10.4081/btvb.2022.57
[This abstract book contains the abstracts presented at the 27th National Congress of the Società Italiana per lo Studio dell'Emostasi e della Trombosi (SISET)]
application/pdf
2785-5309
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https://creativecommons.org/licenses/by-nc/4.0
Bleeding, Thrombosis and Vascular Biology
SISET
27th SISET National Congress | Perugia, 2-5 November 2022
Text.Serial.Journal
https://www.btvb.org/btvb
1
Abstract Book 
2022-10-28
2022-10-28
2022-11-02
2022-11-02
Bleed Thromb Vascul Biol
Società Italiana per lo Studio dell'Emostasi e Trombosi - SISET
2022/10/28
10.4081/btvb.2022.57
2785-5309
s1
SISET
en
.
27th SISET National Congress | Perugia, 2-5 November 2022
https://www.btvb.org/btvb/article/download/57/44
1
SISET_abstrac book_cover dorso 9mm.qxp_Layout 1 27/10/22 08:35 Pagina 1




                                                                                                                                                                      Official Journal of
                                                                                                                               www.btvb.org
                                                                                                                                                                                        SISET
                                                                                                                                                                      Società Italiana per lo Studio dell’Emostasi e della Trombosi




                                                                                                                                               BLEEDING,
                                                                                                                                               THROMBOSIS            AND




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                                                                                                                                               VASCULAR




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                                                                                                                                               BIOLOGY




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                                                                                                                                  XXVII Congresso Nazionale SISET
                                                                                                                                  Perugia | 2-5 novembre 2022




                                                               Official Journal of


                                                                                 SISET
                                                               Società Italiana per lo Studio dell’Emostasi e della Trombosi
                                                                                                                                         btvb
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                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                              Bleeding, Thrombosis and Vascular Biology

                                                                  EDITORIAL BOARD

                                                                    SISET President
                                                               Paolo Gresele, Perugia, Italy


                                                                    Editor-in-Chief
                                  Giovanni de Gaetano, Istituto Neurologico Mediterraneo Neuromed, Pozzilli (IS), Italy


                                                                 Assistant Editor-in-Chief
                                     Chiara Cerletti, Istituto Neurologico Mediterraneo Neuromed, Pozzilli (IS), Italy


                                                                      Section Editors




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                                                             Francesco Bernardi, Ferrara, Italy
                                                            Giancarlo Castaman, Firenze, Italy




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                                                           Raimondo De Cristofaro, Roma, Italy
                                                         Licia Iacoviello, Pozzilli and Varese, Italy
                                                             Gualtiero Palareti, Bologna, Italy




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                                                          Anna Randi, London, United Kingdom
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                                                                Junior Associate Editors
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                                                            Emilia Antonucci, Bologna, Italy
                                                            Alessio Branchini, Ferrara, Italy
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                                                             Simona Costanzo, Pozzilli, Italy
                                                             Stefano Lancellotti, Roma, Italy
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                                                               Silvia Linari, Firenze, Italy
                                                            Roxana Ola, Mannheim, Germany
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                                                                     Board Members
                                     Walter Ageno, Italy                                        Rossella Marcucci, Italy
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                                    Lina Badimon, Spain                                       Maurizio Margaglione, Italy
                                 Stefano Barco, Switzerland                                    Juan Melero-Martin, USA
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                                    Marina Camera, Italy                                          Vittorio Pengo, Italy
                                   Caterina Casari, France                                        Paolo Prandoni, Italy
                               Alessandro Casini, Switzerland                                      Nicola Pozzi, USA
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                                    Sergio Coccheri, Italy                                       Domenico Prisco, Italy
                               Augusto Di Castelnuovo, Italy                                      Alireza Rezaie, USA
                                  Giovanni Di Minno, Italy                                     Marc Righini, Switzerland
                                Maria Benedetta Donati, Italy                                      Bianca Rocca, Italy
                             Karin Fijnvandraat, The Netherlands                              Francesco Rodeghiero, Italy
                                  Alessandro Gialluisi, Italy                                      Luigi Sironi, Italy
                                  Marc Hoylaerts, Belgium                                        David Smadja, France
                                    Paula James, Canada                                        Alessandro Squizzato, Italy
                                  Andrew D. Johnson, USA                                       Saverio Stranges, Canada
                                 Wolfgang Koenig, Germany                                    Hugo Ten Cate, The Netherlands
                                 Maria Elisa Mancuso, Italy                                       Alberto Tosetto, Italy
                              Pier Mannuccio Mannucci, Italy                                      Elena Tremoli, Italy
                                  Lorenzo Mantovani, Italy                                      Armando Tripodi, Italy


                                                                   Managing Editor
                                                                 Emanuela Fusinato, Italy




                                                             [Hematology Reports 2021; 13:s3]                                 [page I]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina II



         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                 SISET - Società Italiana per lo Studio dell'Emostasi e Trombosi

                                                               CONSIGLIO DIRETTIVO

                                                                       Presidente
                                                               Paolo Gresele, Perugia, Italy


                                                                     Vice Presidente
                                                             Valerio De Stefano, Roma, Italy




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                                                                    Past President
                                                              Armando Tripodi, Milano, Italy




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                                                                    Consiglieri
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                                                          Laura Contino, Alessandria, Italy
                                                            Laura Russo, Bergamo, Italy
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                                                        Marco Paolo Donadini, Varese, Italy
                                                  Giovanni Luca Tiscia, San Giovanni Rotondo, Italy
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                                                                      Segretario
                                                              Loredana Bury, Perugia, Italy
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                                                                      XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




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                        The XXVII National Congress of the Italian Society on Hemostasis and Thrombosis (SISET), to be
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                     held in Perugia, November 2-5, 2022 is the first congress in presence since 2018.
                                                                 er


                        SISET was founded 52 years ago and held its first Congress in Parma, in 1970.
                                                           m



                         This will be a four-day important scientific event the motto of which is “Hemostasis: the present and
                     the future”.
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                         This conference will include plenary, educational and state of the art sessions delivered by the most
                     distinguished Italian scientists in the field of hemostasis and thrombosis and by several prominent
                                             -c




                     international scientists. They will present and discuss the most recent findings and the current knowledge
                     on the pathophysiology, diagnostics and treatment of hemorrhagic and thrombotic disorders.
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                         A series of lectures will deal with the new horizons in basic and clinical research including genomics,
                     transcriptomics, artificial intelligence and new drug developments.
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                         A large space will be dedicated to the copious original research produced by Italian scientists in the last
                     year, with many sessions of oral communications and posters, presented by young clinicians and researchers.

                         This year, all these valuable scientific contributions are published in the current special issue of BTVB,
                     the new, open access, international official scientific journal of SISET, with a prestigious international
                     editorial board.
                         BTVB represents a high quality international forum for research on basic, translational and clinical
                     aspects of hemostasis, thrombosis, vascular biology and related fields. We are all proud that the most recent
                     Italian scientific research is now presented in preview in the journal.

                                                                                                                     Paolo Gresele
                                                                                                              President of the SISET




                                                            [Hematology Reports 2021; 13:s3]                                           [page III]
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                                                                                XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                     XXVII Congresso Nazionale SISET
                                                                             Perugia
                                                                       2-5 novembre 2022




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                                                                                INDEX



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                    Selected oral communications ..................................................................................                     1
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                    Oral communications ................................................................................................               11
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                    Poster..........................................................................................................................   81
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                    Index of Authors ........................................................................................................ 165
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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 1



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                         Selected oral communications




                                                                                                           ly
                                                                                                   on
                                                                                   13UOC   Angiologia, Dipatimento di Medicina Clinica,
                    OC001
                                                                                   Azienda ULSS 2Marca Trevigiana, Ospedale San
                    D-DIMER AND REDUCED DOSE APIXABAN FOR
                                                                                   Giacomo Apostolo, Castelfranco Veneto (TV); 14Centro




                                                                                              e
                    EXTENDED TREATMENT AFTER UNPROVOKED                            Emostasi e Trombosi, Dipartimento Emato-Oncologico,
                    VENOUS THROMBOEMBOLISM. THE APIDULCIS
                    STUDY                                                              us
                                                                                   Presidio Ospedaliero deLellis, AzOsp“Pugliese-
                                                                                   Ciaccio”, Catanzaro; 15UOSD di Angiologia, Ospedale
                                                                                   di Conegliano, Conegliano (TV); 16UO Emostasi e
                                                                                   al
                    C. Legnani1, G. Palareti1, D. Poli2, W. Ageno3,                Trombosi, Centro Ricerca Poliambulatorio Giovanni
                    E. Antonucci1, E. Bucherini4, S. Testa5, O. Paoletti5,         Paolo II, IRCCS Ospedale Casa Sollievo della
                                                                           ci

                    A. Chistolini6, A. Serrao6, I. Martinelli7, P. Bucciarelli7,   Sofferenza, San Giovanni Rotondo (FG); 17Clinica
                    A. Falanga8, A. Tosetto9, L. Sarti10, D. Mastroiacovo11,
                                                                     er


                                                                                   Cardiologica, Azienda Ospedaliera di Padova, Padova,
                    B. Cosmi12, A. Visonà13, R.C. Santoro14,                       Italy
                    N. Zanatta15, E. Grandone16, L. Bertù3, V. Pengo17,
                                                               m



                    L. Caiano3, P. Prandoni1, for the Apidulcis study group        Background and Aims: D-dimer assay may be useful
                                                  om




                    1Fondazione                                                    to stratify patients with venous thromboembolism
                                   Arianna Anticoagulazione, Bologna;
                    2Malattie                                                      (VTE) for the risk of recurrence. This approach was
                               Aterotrombotiche, AOU Careggi, Firenze;
                    3UOC Pronto Soccorso, Medicina d’Urgenza e Centro              never evaluated after direct oral anticoagulants are
                                                                                   available. With this multicenter, prospective cohort
                                               -c




                    Trombosi ed Emostasi, ASST dei Sette Laghi, Varese; 4SS
                    Medicina Vascolare e Angiologia, Medicina 2, AUSL              study we wanted to assess the usefulness of an algo-
                                                                                   rithm incorporating serial D-dimer testing to avoid
                                       on




                    Romagna, Ravenna; 5Centro Emostasi e Trombosi,
                    UUOO Laboratorio Analisi chimico-cliniche e                    extended treatment if results are negative and to give
                    microbiologiche,     ASST     Cremona,       Cremona;          apixaban 2.5 mg twice daily if positive. Methods: 732
                                  N




                    6Dipartimento di Medicina Traslazionale e di Precisione        outpatients, from 49 Italian centers, aged 18 to 74 years,
                    Sapienza Università di Roma, Roma; 7Centro Emofilia            anticoagulated for at least 12 months after a first unpro-
                    e Trombosi ABianchi Bonomi, Fondazione IRCCS Ca’               voked VTE, were included. Patients underwent D-
                    Granda, Ospedale Maggiore Policlinico, Milano;                 dimer testing with commercial assays and pre-estab-
                    8Divisione    di Immunoematologia e Medicina                   lished cutoffs. If the first D-dimer, during anticoagula-
                    Trasfusionale & Centro Emostasi e Trombosi, ASST               tion, was negative anticoagulation was stopped and
                    Papa Giovanni XXIII, Bergamo; 9UOC Ematologia,                 testing repeated after 15, 30 and 60 days. At first posi-
                    Centro Malattie Emorragiche e Trombotiche (CMET),              tive result, patients received apixaban for 18 months;
                    AULSS 8Berica Ospedale SBortolo, Vicenza; 10Centro             those with negative results were left without anticoagu-
                    per la diagnosi e la sorveglianza della malattia               lation, and all underwent follow-up for 18 months.
                    tromboembolica, UO Medicina interna d’urgenza,                 Results: D-dimer resulted persistently negative in 286
                    Azienda Ospedaliero Universitaria Policlinico di               patients (39.1%) and positive in the remaining (n=446,
                    Modena, Ospedale Civile Baggiovara, Modena;                    60.9%). During the entire follow-up of 869 years, 26
                    11UOSD     Angiologia e Diagnostica Vascolare,                 primary outcomes occurred in the 732 subjects who
                    Dipartimento Medico, Ospedale Civile SS Filippo e              correctly followed the study protocol (12 proximal
                    Nicola, Avezzano (AQ); 12UO di Angiologia e Malattie           DVTs with or without PE, 10 isolated PEs, and 4 MB
                    della Coagulazione, Azienda Ospedaliero Universitaria          events), for an incidence rate of 3.0% person-years
                    di Bologna, Policlinico SOrsola-Malpighi, Bologna;             (95%CI, 1.9-4.4). The study was prematurely stopped




                                                                [Hematology Reports 2021; 13:s3]                                                [page 1]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 2



         Selected oral communications




                    when a planned interim analysis found an unacceptable       However, thrombocytopenia may also arise from other
                    (according to a pre-established stopping rule) higher       causes than VITT in some patients diagnosed with a
                    prevalence of recurrences in patients who remained off      thrombotic event (TE). Aim of the study was to collect
                    anticoagulation compared to those receiving reduced         relevant information on thrombosis associated with
                    dose apixaban. Twenty-one primary events (19 of effi-       thrombocytopenia occurring after COVID-19 vaccina-
                    cacy) occurred in patients (14 males) with negative D-      tion. Methods: SISET VAX COVID-19 is an ongoing
                    dimer (6.2% person-years, 95% CI, 3.9-9.5), and 5 (3 of     national, multicenter, prospective registry that was set
                    efficacy; 4 males) in those receiving apixaban after pos-   up by the Italian Society on Thrombosis and
                    itive D-dimer results (0.9% person-years; 95% CI, 0.3-      Haemostasis, enrolling consecutive adult patients
                    2.2). The unadjusted HR for primary outcomes between        objectively diagnosed with any TE occurring within
                    the two groups of patients was 6.6 (95% CI, 2.7-19.8),      30 days after receiving any COVID-19 vaccine dose,
                    and the adjusted HR was 8.2 (95% CI, 3.2-25.3). The         with or without thrombocytopenia. Primary objective
                    rate of primary outcomes in patients with negative D-       was the characterization of TEs, in terms of type and
                    dimer was significantly lower (p=0.0362) when the           extension of thrombosis (venous, arterial, microvascu-
                    index event was associated with weak risk factors and       lar), diagnosis, treatment and adverse events (throm-
                    in young (aged <51 years) patients (p=0.0460). Though       bosis progression/early recurrence, major bleeding
                    no other difference reached statistical significance, it    and death) during 30-day follow-up. The study was
                    seems useful to note that recurrences occurred less in      approved by the national ethics committee for studies
                    women whose index event was associated with hor-            on COVID-19 and by participating centers.




                                                                                                        ly
                    monal therapy. Nine events (incidence of 33.3% per-         Fondazione Arianna Anticoagulazione provided sup-
                                                                                port for web-based dataset implementation, data man-




                                                                                                 on
                    son-years) occurred in the 40 subjects who did not cor-
                    rectly follow the algorithm. Conclusions: In patients       agement and approval procedures. Results: From June
                    anticoagulated for >12 months after a first unprovoked      3rd, 2021, to March 20th, 2022, 237 patients from 41
                    VTE, the decision whether to further extend anticoagu-      centers were included in the registry. This report




                                                                                           e
                    lation should not be based on D-dimer testing. Reduced      focuses on 38 patients (16%) who had concomitant
                    dose apixaban is highly effective and safe against recur-
                    rences in patients with positive D-dimer.                       us
                                                                                thrombocytopenia. The mean age was 58 years (stan-
                                                                                dard deviation (SD) 15) and female/male proportion
                                                                                was 18/20. TE occurred after a mean of 11.7 days (SD
                                                                                al
                                                                                7.8) after the latest vaccine dose (30 after 1st or unique
                    OC002                                                       dose, 4 after 2nd dose, 4 after 3rd dose), that was rep-
                                                                        ci

                                                                                resented by ChAdOx1 nCOV-19 (Astrazeneca) in 25
                                                                                patients, Ad26.COV2.S (Janssen) in 2, BNT162b2
                                                                  er


                    THROMBOTIC EVENTS ASSOCIATED WITH                           (Pfizer/BioNTech) in 8 and mRNA-1273 (Moderna) in
                    THROMBOCYTOPENIA AFTER COVID-19                             3. TEs involved a single venous site in 19 patients, a
                                                             m



                    VACCINATION IN ITALY: A REPORT FROM THE                     single arterial site in 6 patients and multiple sites in 13
                    SISET VAX COVID-19 DATABASE                                 patients (4 only venous multiple sites, 9 combined
                                                 om




                                                                                venous and arterial sites). In 22/38 patients (57.8%) a
                    M.P. Donadini1, E. Tarasconi1, L. Bertù1, E. Antonucci2,    diagnosis of VITT was reported, supported in 17
                    L. Contino3, V. De Stefano4, L. Russo5, G. Tiscia6,         patients by positive antibody assay against
                    A. Tripodi7, P. Gresele8
                                              -c




                                                                                PF4/heparin (16 patients) and/or platelet activation
                                                                                assay (6 patients). All these patients had received
                    1Centro
                                      on




                            di ricerche per le malattie tromboemboliche e le    Astrazeneca (21) or Janssen (1) vaccine; 2 patients
                    terapie antitrombotiche, Dipartimento di Medicina e         had been exposed to heparin in the past. TE involved
                    Chirurgia, Università dell’Insubria Varese; 2Fondazione     at least one unusual site in 16/22 patients. The median
                                  N




                    Arianna Anticoagulazione, Bologna; 3S.S.DHaemostasis        platelet count at nadir was 33 × 109/L (Interquartile
                    and Thrombosis Center, Internal Medicine Department,        range 22-50). Patients diagnosed with VITT were
                    “SS Antonio e Biagio” Hospital, Alessandria; 4Sezione di    treated with anticoagulants (16 patients fondaparinux,
                    Ematologia, Università Cattolica, Fondazione Policlinico    6 argatroban and 6 DOAC) and received immunoglob-
                    AGemelli IRCCS, Roma; 5Division of Immunohematology         ulin (21), high-dose steroids (17) and plasma
                    and Transfusion Medicine, Hospital Papa Giovanni XXIII,     exchange (2). (Table 1). During 30-day follow-up, 9
                    Bergamo; 6U.O.SDipartimentale e di Ricerca Emostasi e       patients (40.9%) experienced thrombosis progression
                    Trombosi; Fondazione IRCCS Casa Sollievo della              and 4 (18.2%) died. For the remaining 16/38 patients
                    Sofferenza, San Giovanni Rotondo; 7IRCCS Ca’ Granda         (42.1%), thrombocytopenia was attributed to other
                    Maggiore Hospital, Angelo Bianchi Bonomi Hemophilia         causes than VITT: immune thrombocytopenic purpura
                    and Thrombosis Center, Milano; 8. University of Perugia,    (2), disseminated intravascular coagulation (2), throm-
                    Department of Medicine and Surgery – Section of Internal    botic thrombocytopenic purpura (1), systemic infec-
                    and Cardiovascular Medicine, Italy                          tion (1), myelodysplastic syndrome (1), iatrogenic (1),
                                                                                no specific causes(8). Conclusions: In this national
                    Background and Aims: Vaccine induced immune                 registry on TEs occurring after vaccination against
                    thrombotic thrombocytopenia (VITT) is a very rare           COVID-19, a diagnosis of VITT was extremely rare,
                    and life-threatening complication occurring after ade-      was reported only after adenoviral vector-based
                    noviral vector-based COVID-19 vaccination.                  COVID-19 vaccines in both sexes and was associated




         [page 2]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 3



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    with significant thrombosis progression and mortality.        corrected mice (80%) compared to HA controls (30%).
                    About 40% of reported TEs with thrombocytopenia               None of the transplanted mice developed anti-FVIII or
                    was not attributable to VITT.                                 anti-GFP antibodies. Conclusions: Transplantation of
                                                                                  HSCs from adult BM and FL may provide a novel,
                    Table 1. Treatment of patients with VITT.                     more stable and highly promising preclinical model for
                                                                                  pediatric HA treatment. Our results show FL cells hav-
                                                                                  ing higher engraftment ability, thus paving the way for
                                                                                  studies aimed at maximizing the engraftment and pro-
                                                                                  liferation of donor FVIII-corrective cells while mini-
                                                                                  mizing/avoiding harming pre-conditioning regimens.


                    OC003                                                         OC004


                    TRANSPLANTATION OF FETAL LIVER AND ADULT                      EXPRESSION AND LOCALIZATION OF
                    BONE MARROW CELLS FOR PHENOTYPIC                              COAGULATION FACTOR XI AND ITS RECEPTOR
                    CORRECTION OF HEMOPHILIA A                                    APOER2 IN HUMAN PLATELETS OF HEALTHY
                                                                                  SUBJECTS AND ASSESSEMENT OF ITS




                                                                                                           ly
                    A. Cottonaro1, S. Merlin1, S. Akula1, V. Kalandadze1,         CONTRIBUTION TO PLATELET PROCOAGULANT
                    S. Buzzi1, E. Borroni1, T. Garcia-Leal2, L.J. Serrano2,       ACTIVITY




                                                                                                   on
                    A. Liras3, M.J. Sanchez2, A. Follenzi1
                                                                                  A. Becchetti1, M. Brambilla1, P. Canzano1, M. Conti1,
                    1Department of Health Sciences, University of Piemonte        M. Camera1,2




                                                                                             e
                    Orientale, Novara, Italy; 2Centro Andaluz de Biología
                    del Desarrollo (CABD), CSIC, UPO, Seville, Spain;
                    3Department of Genetic, Physiology and Microbiology,

                    Biology School, Complutense University of Madrid,
                                                                                      us
                                                                                  1Centro  Cardiologico Monzino IRCCS, Milan, Italy;
                                                                                  2Department    of Pharmaceutical Sciences, Università
                                                                                  degli Studi di Milano, Milan, Italy
                                                                                  al
                    Spain
                                                                                  Background and Aim: Coagulation Factor XI (FXI) is
                                                                          ci

                    Background and Aims: We previously showed that                an important member of the contact pathway; its activity
                    adult-derived FVIII-producing cells, i.e. liver sinu-         is critical for thrombus growth and stability, but plays a
                                                                    er


                    soidal endothelial cells (LSECs) and hematopoietic            secondary role in haemostasis. It circulates in plasma and
                    stem cells (HSCs), can be used for the treatment of           binds platelets through Apolipoprotein E Receptor 2
                                                                m



                    adult HA mice. However, after transplantation in busul-       (ApoER2). The binding makes FXI more accessible for
                    fan-conditioned newborn mice, adult LSEC/HSC can-             activation and induces signal amplification that promotes
                                                   om




                    not efficiently engraft compared to murine fetal liver        platelet activation. Furthermore, FXI is stored in platelets
                    (FL) hemato/vascular cells from day 11-13 of gestation        and can be secreted upon activation. Not all platelets
                    (E11-E13) that strongly reconstitute the hematopoietic        have a role in haemostasis: indeed only 30% of platelets
                                                -c




                    compartment and showed multi-organ endothelial                express both phosphatidylserine and Tissue Factor (TF)
                    reconstitution potential and FVIII secretion ability. The     and bind coagulation factors. Thus, the aims of this study
                                        on




                    aim is to investigate the ability of FL cells in repopulat-   were 1) to clarify whether FXI and ApoER2 are present
                    ing the hemato/vascular compartment following trans-          in all platelets or only in a subpopulation and 2) to assess
                    plantation in newborn HA mice without precondition-           intracellular localization through pharmacological
                                   N




                    ing. Methods: We transplanted adult BM or FLE11-              approach; 3) to assess the contribution of platelet-associ-
                    E13 cells from GFP mice into newborn and adult HA             ated FXI to thrombin generation (TG). Methods:
                    mice pre-treated (+BU) or not (noBU) with busulfan.           Platelet-associated FXI and ApoER2 expression were
                    The engraftment level and FVIII activity was assessed         evaluated in 25 healthy subjects by whole blood (WB)
                    starting from 4w after transplantation and followed-up        flow cytometry under basal conditions and upon stimula-
                    for 18m. Bleeding phenotype correction was verified by        tion. Platelet FXI associated thrombin generation (TG)
                    hemarthrosis induction and bleeding assay. Results: In        was assessed by CAT assay. TG was evaluated in pres-
                    all BU-conditioned groups we observed long term               ence and absence of FXI-blocking antibody in FXI
                    (18m) stable GFP+ blood engraftment (>60%) with up            depleted plasma or in plasma pool CTI. Results: Only a
                    to 16% FVIII activity following FL and BM cells trans-        platelet subpopulation expresses FXI on the surface
                    plantation. Interestingly, without pre-conditioning we        (21.4±7.7%) and ApoER2 behaved similarly (26±10%),
                    observed lower but stable engraftment (≤12%) and con-         they colocalized in�15% of platelets. They are also
                    sequent FVIII activity (1-4%) after FL cells transfer,        intracellularly stored in�50% of platelets. Stimulation
                    while BM cells did not engraft in noBU mice. Tail bleed       with different agonists results in a further significant
                    challenge and induced hemarthrosis experiments fur-           increase in the number of positive platelets and in the
                    ther confirmed the phenotypic correction in all mice          amount of antigens/platelet (FXI: ADP, 29±15.7%;
                    receiving FL/BM+BU and in FLnoBU mice.                        TRAP-6, 30.3±14.7%; U46619, 29.4±13.9% p=0.04
                    Additionally, we observed an increased survival rate in       compared to resting condition; ApoER2; ADP,




                                                                [Hematology Reports 2021; 13:s3]                                                 [page 3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 4



         Selected oral communications




                    45.1±15.3%, TRAP-6, 45.1±8.6%, U46619, 48.6±16.7%               in 5/9 patients and >500 BU in two of them. All major
                    p=0.031 compared to resting condition). This upregula-          surgeries were planned to be managed with rFVIIa in
                    tion was blunted when stimulation was performed in the          addition to emicizumab prophylaxis. All patients were
                    presence of the ApoER2 inhibitor RAP and also in the            treated with 2-3 bolus of 90 µg/kg rFVIIa every 3 hours
                    presence Colchicine. This last finding highlights the           at the beginning of surgery till wound suturing, followed
                    localization of both FXI and ApoER2 in the open                 by FVIIa 90 µg/kg every 4 hrs during the first 2 days,
                    canalicular system (OCS). To assess the contribution of         every 6 hrs on days 3-4, every 8 hrs on days 5-7, twice
                    platelet-FXI to TG, we evaluated platelet TG in FXI-            a day on days 8-14 and daily (before the rehabilitation,
                    depleted plasma. TG was delayed compared to autolo-             when required) on days 15-20, then rFVIIa was discon-
                    gous plasma sample (Lagtime: autologous plas-                   tinuated. Adjunctive intravenous tranexamic acid (TA) 1
                    ma:11±2min,         FXI-depleted        plasma17.3±1min;        g every 12 hours for 7 days was also administered.
                    p=0.0022). In the presence of an anti-FXI antibody,             Thromboprophylaxis was not prescribed. Minor surger-
                    Lagtime was further slowed indicating that platelet FXI         ies were managed without rFVIIa prophylaxis, except
                    is functionally active and plays a key role in sustaining       for 2 potentially complicated tooth extractions. A bolus
                    the amplification phase in absence of plasma FXI (base-         of rFVIIa (90 µg/kg) was infused before procedure, fol-
                    line: 17.3±1min, +αFXI Ab 31.5±1min; p=0.0313).                 lowed by oral TA (1 g every 8 hours for 5 days). Results:
                    Moreover, when TG was performed in plasma pool CTI,             No thrombotic events, thrombotic microangiopathy, sig-
                    thus FXI could be activated only by TF-generated throm-         nificant changes of thrombophilia/microangiopathy
                    bin, Lagtime in presence of αFXI antibody was pro-              markers were observed. Bleeding was as expected, apart




                                                                                                            ly
                    longed compared to basal condition (baseline:                   from retroperitoneal toilet, which however was con-
                    23.7±6.6min, +αFXI Ab: 36.4±11min; p=0.05) support-             trolled without additional rFVIIa injections.




                                                                                                     on
                    ing the evidence that FXI has a pivotal role in platelet        Conclusions: Major surgery with a regimen of emi-
                    TG. Similarly, inhibition of ApoER2 also results in a           cizumab and rFVIIa in PWHAi has been safely and effi-
                    decreased TG. Conclusions: All together these data indi-        caciously performed. This represents the first series of




                                                                                               e
                    cate that: 1) FXI and ApoER2 are expressed by a subpop-         major surgery ever reported in this setting using an inter-
                    ulation of platelets and partially co-localized; 2) they are
                    intracellularly stored in OCS; 3) platelet FXI is function-
                    ally active and plays a key role in sustaining the amplifi-
                                                                                        us
                                                                                    nal developed protocol. Minor surgeries can be safely
                                                                                    performed without prophylaxis with rFVIIa. A multidis-
                                                                                    ciplinary team and strict monitoring are recommended
                                                                                    al
                    cation phase of platelet TG.                                    in management of surgical procedures in PWHAi treated
                                                                                    with emicizumab.
                                                                            ci

                                                                                    Table 1.
                    OC005
                                                                     er
                                                               m



                    SURGICAL PROCEDURES IN SUBJECTS WITH
                    HEMOPHILIA A AND INHIBITORS TREATED WITH
                                                   om




                    EMICIZUMAB: A SINGLE CENTRE EXPERIENCE

                    S. Linari1, L. Pieri1, C. Carulli2, F. Demartis1, R. Fjerza1,
                    M.C. Di Stefano1, G. Castaman1
                                                -c




                    1Centre
                                        on




                             for Bleeding Disorders and Coagulation,
                    Department of Oncology, Azienda Ospedaliero-
                    Universitaria Careggi, Florence, Italy; 2Orthopaedic
                                   N




                    Clinic, Orthopedic Traumatologic Centre, Azienda
                    Ospedaliero-Universitaria Careggi, Florence, Italy

                    Background and Aims: The management of surgery in
                    persons with hemophilia A and inhibitors (PWHAi) has
                    always been a major challenge and typically requires the
                    use of activated prothrombin complex concentrates
                                                                                    OC006
                    (aPCC) or recombinant activated FVII (rFVIIa). The
                    recent widespread use of the bi-specific monoclonal
                    antibody emicizumab for long-term prophylaxis in                THE HUMAN NPM1 MUTATION A IMPAIRS
                    PWHAi has required a reassessment of the antihemor-             PROPLATELET FORMATION AND PLATELET
                    rhagic perioperative treatments with by-passing agents.         FUNCTION
                    The aim of this study is to report our experience in man-
                    agement of surgical procedures in PWHAi on emi-                 L. Bury1, S. Momi1, E. Falcinelli1, E. Varasano2,
                    cizumab. Methods: Between January 2018 and April                B. Falini2, P. Sportoletti2, P. Gresele1
                    2022, nine PWHA and high titre inhibitors underwent 31
                    surgical procedures (14 major surgeries, 17 minor sur-          1Section of Internal and Cardiovascular Medicine,

                    geries). At the time of surgery inhibitor titer was >5BU        Department of Medicine and Surgery, University of




         [page 4]                                           [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 5



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Perugia, Perugia, Italy; 2Section of Hematology and          p<0.05). Photochemical damage-induced femoral
                    Clinical Immunology, Centro di Ricerche Emato-               artery thrombosis was significantly delayed, occurring
                    Oncologiche, Department of Medicine and Surgery,             in 11.1±1.2min in control mice and in 30±0.8min in
                    University of Perugia, Perugia, Italy                        Npm1-mutA (p<0.05). Conclusions: Unexpectedly,
                                                                                 given its recognized function of shuttle between
                    Background and Aims: Nucleophosmin (NPM1) is a               nucleus and cytoplasm, platelets express NPM1,
                    multifunctional phosphoprotein which shuttles                which plays a significant role in platelet formation and
                    between the nucleus and the cytoplasm. NPM1 muta-            function. These findings suggest an NPM1-mediated
                    tions, the most common of which is the type A variant,       regulation of genes coding for proteins involved in
                    result in the aberrant localization of the protein in the    several aspects of platelet generation and function,
                    cytoplasm and characterize a specific type of acute          that is lost in the presence of a mutant NPM1, and for
                    myeloid leukemia (AML) reported in 20-30% of                 which further studies are warranted.
                    cases. A conditional mouse model expressing the
                    human NPM1 mutation A in hematopoietic cells pre-
                    viously created did not develop AML and showed               OC007
                    reduction of platelet count, increased mean platelet
                    volume and prolonged bleeding time. In this study we
                    aimed to get a deeper insight into the role of NPM1 in       PLATELETS TRANSFECTED WITH A KRASG12D-
                    megakaryocyte (MK) differentiation, platelet forma-          SILENCING SIRNA REDUCE PANCREATIC TUMOR




                                                                                                         ly
                    tion and function using the conditional mouse model          MASS IN VIVO IN MICE
                    carrying the NPM1 mutation A. Methods: MK were




                                                                                                  on
                    isolated by marrow cells and cultured in the presence        M. Malvestiti, G. Purgatorio, E. Cesari, S. Momi,
                    of TPO. Proplatelet formation was evaluated by fluo-         P. Gresele
                    rescence microscopy. NPM1 was assessed in ultrapu-




                                                                                            e
                    rified control platelets by PCR and Western blotting.        Sezione di Medicina Interna e Cardiovascolare, Dip.
                    Platelet life-span was assessed by flow cytometry
                    upon injecting mice with an anti-GPIX mAb conjugat-
                    ed with DyLight 488. Platelet glycoprotein expression
                                                                                     us
                                                                                 Medicina e Chirurgia, Università di Perugia, Italy

                                                                                 Background and Aims: Pancreatic ductal adenocarci-
                                                                                 noma (PDAC) is one of the deadliest solid cancers with
                                                                                 al
                    was assessed by flow cytometry. P-Selectin and
                    JON/A binding upon platelet activation by convulxin          a dismal 7% 5-year survival rate and is projected to
                                                                         ci

                    (CVX) or thrombin were assessed by flow cytometry.           become the second leading cause of cancer-related mor-
                    Dense granule content and release were assessed by           tality by the end of this decade. Recent exome sequenc-
                                                                   er


                    flow cytometry in platelets labelled with green fluo-        ing studies have provided a detailed genetic profile of
                    rescent mepacrine. Clot retraction was assessed in           PDAC, with mutational activation of the KRAS onco-
                                                             m



                    PRP additioned with thrombin. Thrombus formation             gene found in ~95% of patients. In addition, there is
                    was evaluated in vitro on a collagen-coated surface          compelling evidence that aberrant KRAS protein func-
                                                 om




                    under flow. Tail bleeding time and photochemically-          tion is critical for PDAC growth and progression. The
                    induced femoral artery thrombosis were also studied          extraordinarily complex PDAC tumor microenviron-
                    in vivo. Results: Ultrapurified control human and            ment (TME) is believed to be a crucial cause of drug
                                                                                 resistance and the major hurdle to overcome for any
                                              -c




                    mouse platelets express NPM1 protein but not NPM1
                    mRNA which is found in MK. The number of circulat-           PDAC therapeutic. Moreover, no effective drugs target-
                                       on




                    ing platelets was significantly lower and platelets were     ing KRAS have entered clinical use so far. Small-inter-
                    larger in Npm1-mutA mice compared to control mice            fering RNAs (siRNAs) harbor tremendous therapeutic
                    (227±41x103/μl vs 696±76x103/μl; mean platelet               potential because they offer highly-selective, reversible
                                  N




                    diameter 3.7±1 μm vs 2.9±0.6 μm;p<0.05). The per-            control of gene expression thus potentially silencing
                    centage of MK differentiated from bone marrow cells          pathogenic oncogenes. However, utilization of siRNAs
                    was comparable in Npm1-mutA and control mice,                in vivo has been limited by their short circulating half-
                    however MK from Npm1-mutA mice showed strongly               life due to degradation by serum nucleases and limited
                    defective proplatelet formation. Surface expression of       tumor uptake. A crosstalk between blood platelets and
                    platelet glycoproteins αIIbβ3 and GPVI was reduced,          pancreatic cancer cells has been previously character-
                    expression of CD9 was increased in Npm1-mutA                 ized, thus aim of our study was to verify whether
                    mice. A significant impairment of JON/A binding and          platelets can efficiently deliver therapeutic siRNAs to
                    P-selectin expression was found upon activation,             PDAC TME in vitro and in vivo. Methods: We co-incu-
                    showing defective integrin αIIbβ3 activation and α-          bated human platelets transfected with KRASG12D
                    granules release. Also, secretion of d-granules was          siRNA with Panc-1 cells (PDAC cell line) for 24 and 48
                    impaired. Clot retraction was significantly reduced          hours in order to assess the target gene silencing in Panc-
                    (~60%). Perfusion of Npm1-mutA blood over a colla-           1 cells by q-PCR analysis. Then, human KRASG12D-
                    gen-coated surface revealed that the total surface area      siRNA-transfected platelets were intravenously infused
                    covered by platelets was significantly decreased com-        in NSG (immunotolerant of human cells) mice 7 days
                    pared with wild type mice, both at high and low shear        after the subcutaneous injection of 1x106 Human Panc-
                    rates. Tail bleeding time was slightly, but significantly,   1 cells. A control group of mice infused with non-target-
                    prolonged (610±208 sec vs controls 200±89 sec,               ing scrambled-siRNA (NC) transfected platelets was




                                                              [Hematology Reports 2021; 13:s3]                                                 [page 5]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 6



         Selected oral communications




                    studied in parallel. A total of 6 siRNA-transfected         Sciences “Mario Serio”, University of Firenze,
                    platelet infusions were administered every 2 days start-    Firenze, Italy; 2Department of Experimental and
                    ing from day 7 to day 20 in each mouse. 45x106 human        Clinical Medicine, University of Firenze, Firenze, Italy;
                    siRNA-transfected platelets were transfused each time.      3Clinical   Immunology, Allergy and Advanced
                    Human platelets were isolated from 6 different healthy      Biotechnologies Unit, Azienda Unità Sanitaria Locale-
                    donors. Tumor burden was quantified following animal        IRCCS di Reggio Emilia, Reggio Emilia, Italy;
                                                                                4Department of Biomedical, Experimental and Clinical
                    sacrifice at day 20. During the experiment human
                    platelet count in mouse blood, by measuring the fraction    Sciences “Mario Serio,” University of Firenze,
                    of circulating platelets expressing human CD41, and         Firenze, Italy; Nephrology and Dialysis Unit,
                    animal weight were constantly monitored. Results: In        Department of Pediatrics, Meyer Children’s Hospital,
                    vitro a strong reduction of KRASG12D expression in          Firenze, Italy
                    Panc-1 cells (-92.86±0.11% vs control) after co-incuba-
                    tion for 48 hours with KRASG12D-siRNA-transfected           Background and Aims: Cardiovascular events are
                    platelets was obtained while a non-significant reduction    the major determinant of morbidity and mortality in
                    was found after 24 hours. KRASG12D-siRNA-trans-             systemic lupus erythematosus (SLE), particularly in
                    fected platelets-infused mice showed a reduced tumor        patients with lupus nephritis. A number of studies
                    burden compared with the NC-transfected platelet-           suggested a pivotal role for oxidation in promoting
                    infused group, as shown by a statistically significant      vascular and renal damage in SLE, though the precise
                    decrease of tumor volume and tumor weight (Figure 1).       mechanisms are unclear. This study aimed therefore




                                                                                                       ly
                    No significant differences in the two experimental          to investigate the potential role of oxidative-mediated
                                                                                structural/functional fibrinogen alterations in the




                                                                                                on
                    groups were found regarding body weights and circulat-
                    ing human transfected platelets (3.18±0.83% vs              pathogenesis of atherothrombosis in SLE. Methods:
                    3.99±0.51% at day 11). Conclusions: Our data show           We enrolled 144 adult SLE patients and 90 matched
                    that platelets transfected with a siRNA targeting           controls. We assessed specific plasma oxidative stress




                                                                                          e
                    KRASG12D and co-incubated with PDAC cells in vitro          markers and, in purified fibrinogen fractions obtained
                    are able to strongly silence the tumor cell KRAS onco-
                    gene. The repeated transfusion of human siRNA-trans-
                    fected platelets in mice in which PDAC was subcuta-
                                                                                   us
                                                                                from SLE patients and control subjects, both fibrino-
                                                                                gen oxidative modifications and its functional/struc-
                                                                                tural parameters. In particular, thrombin-catalyzed
                                                                                al
                    neously injected significantly reduced tumor progres-       fibrin polymerization and fibrin resistance to plas-
                    sion. Platelet transfected with siRNAs targeting specific   min-induced lysis were evaluated. Fiber diameter and
                                                                        ci

                    oncogenes may represent an innovative approach for          clot porosity were assessed by stimulated emission
                    chemotherapy resistant cancers.                             depletion (STED) super-resolution microscopy.
                                                                  er


                                                                                Moreover, NADPH oxidase expression in renal biop-
                                                                                sies from patients with active proliferative lupus
                                                             m



                                                                                nephritis was estimated by confocal microscopy.
                                                                                Results: Compared to controls, SLE patients exhibit-
                                                 om




                                                                                ed an increased lymphocyte, monocyte and neutrophil
                                                                                reactive oxygen species (ROS) production, mainly
                                                                                due to neutrophil NADPH oxidase (42432±18998 vs
                                              -c




                                                                                13234±4223 RFU/s, p=0.001), paralleled by an
                                                                                increase in plasma lipid peroxidation, a reduction in
                                      on




                                                                                antioxidant defenses, and an enhanced fibrinogen oxi-
                                                                                dation (294 (256-329) vs 163 (121-184) RFU, p
                                                                                <0.0001). SLE patients also exhibited marked alter-
                                  N




                                                                                ations in fibrinogen structure and clot architecture:
                                                                                this latter was mainly characterized by reduced poros-
                    Figure 1.                                                   ity and by a tight fibrin network with filaments of
                                                                                decreased average size. Furthermore, significant
                                                                                alterations in fibrinogen functional features, namely
                    OC008                                                       thrombin-catalyzed fibrin polymerization and fibrin
                                                                                susceptibility to plasmin-induced lysis were more
                                                                                evident in SLE than in controls. Interestingly, fibrino-
                    OXIDATIVE-MEDIATED STRUCTURAL FIBRINOGEN                    gen structural and functional modifications signifi-
                    MODIFICATIONS: A NEW LINK BETWEEN                           cantly correlated with redox parameters, which in
                    IMMUNITY AND ATHEROTHROMBOSIS IN                            turn correlated with SLE disease activity. NADPH
                    SYSTEMIC LUPUS ERYTHEMATOSUS                                oxidase subunit p22phox was overexpressed in renal
                                                                                biopsies from patients with active proliferative lupus
                    M. Becatti1, G. Emmi2, F.R. Argento1, E. Fini1,             nephritis. Conclusions: collectively, these data sug-
                    A. Mannucci1, A. Bettiol2, M. Nicastro3, A. Vaglio4,        gest that ROS induce fibrinogen structural/functional
                    N. Taddei1, C. Fiorillo1, D. Prisco2                        modifications in SLE; this may represent a new
                                                                                pathogenetic mechanism underlying atherothrombo-
                    1Department   of Experimental and Clinical Biomedical       sis in SLE.




         [page 6]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 7



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    OC009                                                       than 100k/µl and 9.9% lower than 150k/µl. Women
                                                                                were 11.2% of patients with <150k/ul platelets and
                                                                                17.2% of those with <100k/µl versus 28.2% of the
                    LOW PLATELET COUNT HAS A STRONG                             population with normal platelets. Patients with
                    WORSENING IMPACT ON OUTCOME IN PATIENTS                     <150k/µl platelets were older (71±10.9 vs 65.6±12.5
                    WITH ACUTE CORONARY SYNDROMES: FROM                         years, p<0.0001) and had higher incidence of non-
                    THE START ANTIPLATELET REGISTRY.                            valvular atrial fibrillation (17.1%vs 8.6%, p=0.02),
                                                                                previous ischemic stroke (22.5% vs 8.7%, p=0.01) and
                    G. Guglielmini1, M. Del Pinto2, P. Calabrò3,                previous PCI (14.6% vs 7.8%, p=0.001) than patients
                    P. Pignatelli4, G. Patti5, V. Pengo6, E. Antonucci7,        with a normal platelet count. At hospitalization
                    P. Cirillo8, T. Fierro1, G. Palareti7, R. Marcucci9,        patients with <150k/µl had significantly lower hemo-
                    P. Gresele1 and the START Antiplatelet Registry Group       globin levels (12.9±2.1 vs 13.6±1.9 g/dL, p<0.001),
                                                                                hematocrit (38.6±6% vs 40.8±5.4%, p<0.0001) and
                    1Department    of Medicine, Section of Internal and         glomerular filtration rate (73.9±41.2 vs 86.5±32.9
                    Cardiovascular Medicine, University of Perugia;             ml/min, p=0.001). Clinical presentation and type of
                    2Division of Cardiology, Perugia Hospital; 3Department
                                                                                revascularization were similar in normal and low
                    of Translational Medical Sciences University of             platelet count patients, however the latter were dis-
                    Campania “Luigi Vanvitelli”, Naples; 4Department of         charged more frequently on single antiplatelet therapy
                    Clinical,     Internistic,    Anesthesiologic      and      (15.2% vs 8.9%, p=0.028). After two years follow-up,




                                                                                                        ly
                    Cardiovascular Sciences, Sapienza University of Rome;       MACCE-free survival time was significantly shorter
                    5Department of Traslational Medicine, University of




                                                                                                on
                                                                                in patients with platelet count <100k/µl (13.2 vs 21.0
                    Eastern Piedmont, Maggiore della Carità Hospital,           months, p=0.005; HR=9.96, 95% CIs 2 to 49.1) as
                    Novara; 6Department of Cardiac, Thoracic, Vascular          well as for patients with platelet count <150k/µl (17.7
                    Sciences and Public Health, University of Padova;           vs 21.0 months, p=0.001; HR=2.9, 95% CIs 1.5 to




                                                                                           e
                    7Arianna Anticoagulazione Foundation, Bologna;
                                                                                5.5). Also NACE occurred significantly more fre-
                    8Division of Cardiology, Department of Advanced

                    Biomedical Sciences, “Federico II” University, Naples;
                    9Department of Experimental and Clinical Medicine,
                                                                                    us
                                                                                quently in patients with low platelet count
                                                                                (<150k/µl=15.2% vs 8.4%, p=0.02). Conclusions:
                                                                                Low platelet count identifies a subgroup of ACS
                    University of Florence; Italy
                                                                                al
                                                                                patients at significantly increased risk of adverse out-
                                                                                come. Patients admitted to CCU for ACS with a low
                                                                        ci

                    Background and Aims: Some previous observations             platelet count should be treated with special care to
                    suggest that a low platelet count is associated with an     prevent excess adverse outcome.
                                                                  er


                    increased risk of adverse outcome in patients with
                    acute coronary syndromes (ACS). However, most of
                                                             m



                    the information comes from post-hoc analyses of ran-        OC010
                    domized clinical trials, many of which performed sev-
                                                 om




                    eral years ago. Objectives: Our aim was to assess the
                    impact of low platelet count on cardiovascular out-         A FACTOR X VARIANT WITH ENGINEERED
                    come and treatment decisions in patients hospitalized       ACTIVATION PEPTIDE AS INNOVATIVE
                                              -c




                    for ACS in a current real-life setting in Italy. Methods:   BY-PASSING AGENT FOR HAEMOPHILIA
                    START-ANTIPLATELET is a multicenter registry
                                      on




                    enrolling patients admitted to Italian coronary care        E. Tonetto, R. Tarantino, M.F. Testa, L. Peretto,
                    units (CCU) for ACS and treated by PCI or medical           F. Bernardi, M. Pinotti, A. Branchini
                    therapy. Baseline clinical characteristics and treatment
                                  N




                    at discharge were recorded and follow-up reassess-          Department of Life Sciences and Biotechnology,
                    ments were repeated at 6-months, one year and yearly        University of Ferrara, Ferrara, Italy
                    thereafter. Low platelet count was defined as a platelet
                    count at admission lower than 150k/µL. Major cardio-        Background and Aims: In haemophilia with
                    vascular and cerebrovascular adverse events                 inhibitors, by-passing agents hold the promise of
                    (MACCE) were defined as cardiovascular death, non-          enhancing thrombin generation by increasing the con-
                    fatal MI, TIA and ischemic stroke, target vessel revas-     version of FX to its activated form FXa. However,
                    cularization (TVR) and major arterial ischemic events.      FXa, which should be the ideal candidate for therapeu-
                    Net adverse cardiac and cerebrovascular events              tic purposes, is rapidly inhibited and displays very
                    (NACE) were MACCE plus major bleeding. Data                 short half-life in vivo. Hence, the availability of a by-
                    were analyzed using the IBM SPSS 25 software, cate-         passing FX molecule would be of great relevance, with
                    gorical variables were compared using the Fisher            a theoretically low immunogenicity because of the
                    exact test while continuous variables were compared         presence of circulating FX in patients. The aim of this
                    with the t-test or with the Mann-Whitney U test, as         study is to develop a novel hyper-activable engineered
                    appropriate. Clinical event rates were analyzed with        FX variant with by-passing activity (FXHA-BP, patent
                    the Kaplan-Meier method and compared with Logrank           application in progress) able to correct the defective
                    statistics. Results: Among 1890 enrolled patients pre-      intrinsic pathway in haemophilia plasma with
                    senting with an ACS, 1.9% had a platelet count lower        inhibitors. Methods: FX activation peptide (AP) was




                                                             [Hematology Reports 2021; 13:s3]                                               [page 7]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 8



         Selected oral communications




                    rationally engineered by site-directed mutagenesis of        Surgery, G.B. Rossi Hospital, Verona, Italy; 5.
                    wild-type FX (FXwt) and the resulting engineered             Department of Medicine, University of Verona, Italy
                    variants (FXHA-BP-1-4) were transiently expressed in
                    Human embryonic Kidney (HEK) 293 cells. Secreted             Background and Aims: Patients with pancreatic can-
                    FX protein levels were evaluated by ELISA. Activity          cer are at very high risk of venous thromboembolism
                    was assessed by PT-based (FX-deficient plasma) and           (VTE). Activation of the contact system and the intrin-
                    aPTT-based (haemophilia A plasma) assays. Results:           sic pathway may play an important role, but their exact
                    The FXHA-BP variants were transiently transfected in         contribution to hypercoagulability in pancreatic cancer
                    HEK293T and the antigen levels assessed by ELISA.            remains uncertain. We aim to assess the association
                    The FXHA-BP variants showed appreciable secreted lev-        between intrinsic pathway activation as measured by
                    els, with higher values for FXHA-BP3 (65% of FXwt)           complexes of the contact system enzymes with their
                    and FXHA-BP2 (40%) than FXHA-BP1 and FXHA-BP4                serpins and VTE risk in patients with pancreatic cancer.
                    (20%). Intriguingly, all FX variants efficiently short-      Methods: Patients with advanced or metastastic pan-
                    ened coagulation times in PT-based assays in FX-defi-        creatic cancer who were scheduled for palliative
                    cient plasma and showed a specific activity                  chemotherapy were included. Blood was drawn at base-
                    (activity/antigen ratio) slightly increased (~1.6, FXHA-     line and patients were followed for a maximum of 180
                    BP2 and FXHA-BP3; ~2.5, FXHA-BP1 and FXHA-BP4), as           days. The main outcome was VTE, composing of deep-
                    compared to FXwt. Upon intrinsic activation in aPTT-         vein thrombosis of upper or lower extremities, pul-
                    based assays, addition of FXHA-BP1-3 variants to             monary embolism, and splanchnic vein thrombosis.




                                                                                                           ly
                    haemophilia A plasma resulted in a graded shortening         Contact system complexes (PKa:C1, FXIIa:C1,
                    of coagulation times (FXHA-BP1, 155±1 sec; FXHA-BP2,         FXIa:C1, FXIa:α1at, FXIa:AT, and FIXa:AT) were




                                                                                                   on
                    150±2 sec; FXHA-BP3, 151±5 sec), while addition of           measured with ELISA and as well as known biomarkers
                    equal amounts of FXwt resulted in negligible effects         for VTE. Discrimination of complexes and biomarkers
                    (165 sec) as also observed for the negative control          was assessed by areas under the receiver operating




                                                                                             e
                    (medium from untransfected cells) used as baseline.          characteristic curve (AUROC) and subdistribution haz-
                    Noticeably, the FXHA-BP4 variant efficiently restored
                    coagulation, with coagulation times (124±3 sec) signif-
                    icantly different from those of the other FXHA-BP1-3
                                                                                     us
                                                                                 ard ratios (SHR), with death not related to VTE as a
                                                                                 competing risk. Results: Ninety-three patients with
                                                                                 complete 6-month follow-up were included. The mean
                                                                                 age was 66 years (7.8 SD) and 50 (53.8%) were male.
                                                                                 al
                    variants as well as of control FXwt and baseline. These
                    data demonstrated the by-passing activity of the FXHA-       Sixteen (17.2%) patients developed VTE. The AUROC
                                                                         ci

                    BP4 variant even at the relatively low concentrations        was highest for FXIa:α1at (AUC 0.75; 95% CI 0.62-
                    obtained after transient expression. Conclusions: The        0.87) (Table 1). When contrasting the upper quartile
                                                                   er


                    approach of rational engineering on FX AP resulted in        with lower quartiles, the following biomarkers of the
                    FX variants with improved functional properties, with        intrinsic pathway were significantly associated with
                                                             m



                    the relevant ability of by-passing the defective intrinsic   VTE in univariable analyses: FXIa:α1at (SHR 3.53;
                    pathway in haemophilia plasma. In particular, the            95% CI 1.34-9.30) and FXIa:AT (SHR 3.62; 95% CI
                                                 om




                    FXHA-BP4 variant, the most promising by-passing mol-         1.38-9.51). Log-transformed FXIa:α1at (SHR 1.51;
                    ecule, will be further engineered to further improve its     95% CI 1.05-2.18) was also significantly associated
                    by-passing activity as well as its half-life, with the aim   with VTE when assessed as a continuous variable
                                                                                 (Table 1). Conclusions: Markers of contact pathway
                                              -c




                    of developing an extended half-life by-passing variant
                    that might represent a novel therapeutic tool for            activation, such as FXIa:α1at and FXIa:AT complexes,
                                                                                 were strongly associated with VTE risk in patients with
                                       on




                    haemophilia with inhibitors.
                                                                                 pancreatic cancer. These results help to provide insight
                                                                                 in the mechanism of thrombosis in pancreatic cancer
                                  N




                                                                                 and may aid in refining VTE prediction.
                    OC011
                                                                                 Table 1. Competing risk analysis for all venous thromboembol-
                    CONTACT PATHWAY ACTIVATION AND RISK OF                       ic events versus no events during six months of follow-up, with
                    VENOUS THROMBOEMBOLISM IN PATIENTS                           death as a competing risk.
                    WITH PANCREATIC CANCER: THE PROSPECTIVE
                    SENECA STUDY

                    E. Campello1, F.T.M. Bosch2, C. Bulato1, S. Gavasso1,
                    M.W. Henderson3, C. Bassi4, N. Martinelli5, N. van Es2,
                    A. Ilich3, H.R. Büller2, N. Key3, P. Simioni1
                    1.
                     Department of Medicine, University of Padova, Italy;
                    2. Department of Vascular Medicine, Amsterdam
                    University Medical Center, Amsterdam, The
                    Netherlands; 3. UNC Blood Research Center,
                    University of North Carolina at Chapel Hill, Chapel
                    Hill, NC, USA; 4. Unit of General and Pancreatic




         [page 8]                                         [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 9



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    OC012                                                      parameters, no patient was of Asian ethnicity. We failed
                                                                               to confirm the role of body mass index ≥25 kg/m2, cen-
                                                                               tral venous catheter, low-dose Dexamethasone (DEX),
                    IMPACT OF INHERITED AND ACQUIRED                           erythropoietin stimulating agent and therapeutic antico-
                    THROMBOPHILIA ON THE THROMBOTIC RISK IN                    agulation as risk factors for VTE; we confirmed the
                    MULTIPLE MYELOMA                                           prognostic impact of IMIDs, pelvic/hip/femur fractures,
                                                                               doxorubicin, high dose DEX, personal history of VTE
                    F. Di Landro, T. Za, F. Bartolomei, A. Ciminello,          and antithrombotic prophylaxis. Family history of VTE
                    S. Betti, S. Ceglie, F. Ramundo, P. Arcuri, L. Fianchi,    and acquired thrombophilia were significant risk factors
                    L. Pagano, E. Rossi, V. De Stefano                         for VTE; inherited thrombophilia did not enhance the
                                                                               risk of VTE (Table 1). Then we considered the pts with
                    Sezione di Ematologia, Dipartimento di Scienze             an observation time >12 months (N=236): a personal
                    Radiologiche ed Ematologiche, Università Cattolica,        history of VTE and mild inherited thrombophilia (i.e.,
                    Fondazione Policlinico AGemelli IRCCS, Roma, Italy         heterozygous FVL and FII G/A) were independent risk
                                                                               factors for VTE with an hazard ratio (HR) HR of 4.5
                    Background and Aims: Venous thromboembolism                and 5.7, respectively. From this population, we further
                    (VTE) is a common complication of multiple myeloma         analyzed only the pts with responding MM (N=170),
                    (MM). The IMPEDE VTE score is a validated risk             and we recorded 9 VTE events; mild thrombophilia was
                    assessment model for prophylaxis; however, despite         confirmed as a risk factor for VTE (HR 16.2) (Table 1).




                                                                                                      ly
                    antithrombotic prophylaxis, VTE incidence in MM pts        Conclusions: The risk of thrombosis is enhanced dur-
                    remains high, prompting the need for a more tailored       ing the first year from diagnosis of MM by the follow-




                                                                                               on
                    antithrombotic strategy. In this monocentric retrospec-    ing factors: IMIDs, pelvic/hip/femur fractures, doxoru-
                    tive study, we analyzed the impact on VTE risk of the      bicin, high dose DEX, familial and personal history of
                    IMPEDE VTE variables; moreover, we included family         VTE, and acquired thrombophilia. After the first year




                                                                                          e
                    history of VTE and inherited or acquired thrombophilia     from diagnosis, heterozygous FVL and FII G/A and
                    as additional risk factors. Methods: We included in the
                    study only MM pts investigated for inherited and
                    acquired thrombophilia. We considered major VTE as
                                                                                   us
                                                                               personal history of VTE were independent risk factors
                                                                               for VTE. Considering only pts with responding MM,
                                                                               mild inherited thrombophilia was confirmed to be a
                    the primary outcome: deep vein thrombosis, pulmonary
                                                                               al
                                                                               strong risk factor for thrombosis. We suggest that dur-
                    embolism, and cerebral and splanchnic vein thrombo-        ing the first phase of active disease, the moderate risk
                    sis. We analyzed the parameters included in the
                                                                        ci

                                                                               associated with inherited thrombophilia was obscured
                    IMPEDE VTE score, including as additional risk fac-        by the stronger clinical or biochemical risk factors
                                                                  er


                    tors for VTE a family history of VTE, inherited throm-     dependent on the MM burden disease. Conversely, the
                    bophilia (i.e., antithrombin, protein C and S deficien-    constitutional risk due to inherited factors becomes
                    cies, factor V Leiden [FVL], and prothrombin
                                                             m



                                                                               more critical during a stable phase of the disease.
                    G20210A mutation [FII GA]), and acquired throm-
                    bophilic alterations (i.e., hyperhomocysteinemia,
                                                 om




                                                                               Table 1.
                    antiphospholipids, and transitory disease-related defi-
                    ciency of natural anticoagulants). We included as candi-
                    date predictors for VTE in the multivariate Cox propor-
                                              -c




                    tional-hazards regression model only the covariates
                    with a significance level of p<0.1 at the univariate
                                       on




                    analysis. Results: We investigated 292 MM pts (M/F
                    163/129, median age 65 years, range 28-89). During the
                    induction phase (first 12 months from diagnosis), 182
                                  N




                    pts received immunomodulatory agents (IMIDs). We
                    recorded 41 VTE events. Out of the IMPEDE VTE




                                                             [Hematology Reports 2021; 13:s3]                                             [page 9]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 10



         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                                         ly
                                                                                         on
                                                                                 e
                                                                            us
                                                                      al
                                                                  ci
                                                             er
                                                          m
                                              om
                                           -c
                                     on
                                 N




         [page 10]                                    [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 11



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                               Oral communications




                                                                                                         ly
                                                                                                  on
                    OC013                                                         age- and sex-matched patients hospitalized in ICU not
                                                                                  for COVID-19 (non COVID-19 ICU). NO and ROS gen-




                                                                                            e
                                                                                  eration by platelets was assessed by flow cytometry
                    DEFECTIVE NITRIC OXIDE PRODUCTION AND
                                                                                  using two specific fluorescent probes, DAF-FM diacetate
                    INCREASED REACTIVE OXIGEN SPECIES
                    GENERATION BY PLATELETS FROM COVID-19
                    PATIENTS
                                                                                     us
                                                                                  (PMID: 34233447) and DCF (PMID: 26197385), nor-
                                                                                  malizing the fluorescence intensity for platelet number.
                                                                                  Results: Platelets from HS and non COVID-19 ICU
                                                                                  al
                                                                                  patients showed a dose-dependent and comparable
                    E. Petito1, G. Guglielmini1, E. De Robertis2, E. Giglio1,     increase of NO production in response to collagen
                                                                          ci

                    C. Becattini1, E. Falcinelli1, L. Franco1, L. Saccarelli2,    (Figure 1A). On the contrary, platelets from COVID-19
                    F. Gori3, G. Vaudo4, V. Cerotto5, U. Paliani6, P. Gresele1
                                                                    er


                                                                                  patients, both ICU and non ICU, did not show any
                    1Section                                                      increase of NO production in response to increasing col-
                              of Internal and Cardiovascular Medicine;
                                                              m



                    2Division                                                     lagen concentrations, showing an almost complete sup-
                              of Anaesthesia, Analgesia, and Intensive Care,      pression of platelet NO production (Figure 1A). This
                    3Section of Anesthesia, Intensive Care, and Pain Medicine,
                                                  om




                                                                                  suppression appears to be selective to COVID-19
                    Department of Medicine and Surgery, University of
                                                                                  patients, because NO generation by platelets from criti-
                    Perugia, Perugia, Italy; 4Unit of Internal Medicine, Terni
                                                                                  cally ill patients hospitalized in ICU not for COVID-19
                    University Hospital; 5Section of Anesthesia, Intensive Care
                                                                                  did not differ from that of HS (Figure 1A). Moreover,
                                               -c




                    and Pain Medicine, Department of Emergency and
                    Urgency; 6Division of Internal Medicine, ASL 1Umbria,         platelets from COVID-19 patients showed a significant
                                                                                  increase in ROS production compared with HS platelets
                                       on




                    06012, Città di Castello, Italy
                                                                                  but this increase was also evident in non COVID-19 ICU
                    Background and Aims: Oxidative stress, an imbalance           patients (Figure 1B). Conclusions: Our study shows a
                                  N




                    between reactive oxygen/nitrogen species (ROS/RNS)            strikingly defective NO production by platelets from
                    and the antioxidant potential, contributes along with the     COVID-19 patients, independent from disease severity.
                    cytokine storm to COVID-19 pathogenesis (PMID:                Platelets from non COVID-19 ICU patients instead
                    34836751). A hyperactivated platelet phenotype is             released NO normally, and significantly more than
                    strongly involved in the thrombotic complications of          platelets from COVID-19 patients, thus showing a selec-
                    COVID-19 (PMID: 35027697). ROS and NO produced                tive SARS-CoV-2 infection-related defect of platelet NO
                    by activated platelets play a pivotal and opposite role in    production. We also show an increase in ROS production
                    regulating platelet function: ROS potentiate (PMID:           by COVID-19 platelets, evident with the lowest collagen
                    32660144), while NO suppresses platelet activation            dose, indicating that COVID-19 platelets are in a hyper-
                    (PMID: 17901370). To our knowledge no studies have            reactive state independently from disease severity.
                    assessed platelet ROS and NO production in COVID-19.          However, for this parameter platelet hyperreactivity was
                    Our aim was to analyze platelet ROS and NO production         also evident in platelets of patients hospitalized in ICU
                    in COVID-19 patients, comparing it to healthy controls        for reasons different from COVID-19, suggesting that
                    and critically ill non COVID-19 patients. Methods:            the acute inflammatory reaction of the critically ill
                    Venous blood was collected from 24 COVID-19 patients          patient associates with enhanced platelet ROS produc-
                    hospitalized in ordinary wards (non ICU), 24 age- and         tion. Platelets may play a central role in the previously
                    sex-matched healthy subjects (HS), 11 COVID-19                reported decreased NO bioavailability in COVID-19
                    patients hospitalized in intensive care unit (ICU), and 21    (PMID: 34020311). Therapeutic restoration of platelet




                                                               [Hematology Reports 2021; 13:s3]                                               [page 11]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 12



         Oral communications




                     NO production may represent a target for treatments                 biosynthesis and under laminar flow platelet-dependent
                     potentially leading to better clinical outcome in COVID-            thrombus growth in SARS-CoV-2 compared to con-
                     19 patients.                                                        trols; both effects were lowered by Nox2 and Toll-like
                                                                                         receptor 4 (TLR4) inhibitors. Two hours after adminis-
                                                                                         tration of monoclonal antibodies a significant inhibition
                                                                                         of PA was observed in SARS-CoV-2 patients compared
                                                                                         to untreated ones. A docking simulation analysis sug-
                                                                                         gested that TLR4 binds to S protein via three receptor-
                                                                                         binding domains; furthermore, immunoprecipitation
                                                                                         and immunofluorescence showed S protein-TLR4 co-
                                                                                         localization in platelets from SARS-CoV-2. Plasma
                     Figure 1. Nitric oxide (A) and ROS production (B) by resting        from SARS-CoV-2 patients enhanced PA and Nox2-
                     and activated platelets. (*p<0.05 vs their own basal; # p<0.05 vs   related oxidative stress, an effect blunted by TNFα
                     their own Coll 1 µg/mL; § p<0.05 vs their own Coll 5 µg/mL).
                                                                                         inhibitor; this effect was recapitulated by an in vitro
                     Lines report slopes of the concentration-response curves.
                                                                                         study documenting that TNFα alone promoted PA and
                                                                                         amplified the platelet response to S protein via p47phox
                                                                                         up-regulation. Conclusions: The study identifies two
                                                                                         TLR4-dependent and independent pathways promoting
                     OC014
                                                                                         platelet-dependent thrombus growth and suggests inhi-




                                                                                                                ly
                                                                                         bition of TLR4 or p47phox as a tool to counteract
                     TOLL-LIKE RECEPTOR 4-DEPENDENT AND




                                                                                                         on
                                                                                         thrombosis in SARS-CoV-2.
                     INDEPENDENT PLATELET-DEPENDENT
                     THROMBOSIS IN SARS-COV-2 INFECTION
                                                                                         OC015




                                                                                                   e
                     V. Cammisotto1, R. Carnevale2,3, S. Bartimoccia2,
                     C. Nocella1, V. Castellani4, L. Loffredo1, S. Sciarretta2,6,
                     A. Coluccia5, S. Romano5, G. Ceccarelli7, A. Oliva7,
                     M. Venditti7, F. Pugliese4, C. Mastroianni7,
                                                                                            us
                                                                                         TF-POSITIVE SMALL EXTRACELLULAR VESICLES
                                                                                         CONTRIBUTE TO THE PROCOAGULANT
                                                                                         PHENOTYPE IN COVID-19 PATIENTS
                                                                                         al
                     O. Turriziani8, M. Leopizzi1, G. D’Amati9,
                     P. Pignatelli1,3, F. Violi1,3
                                                                                         P. Canzano1, M. Brambilla1, R. Frigerio2, M. Cretich2,
                                                                                ci

                     1Department   of Clinical Internal, Anaesthesiological              A. Becchetti1, M. Conti1, M. Camera1,3
                                                                          er


                     and Cardiovascular Sciences, Sapienza University of                 1Centro
                     Rome; 2Department of Medical-Surgical Sciences and                           Cardiologico Monzino, IRCCS; 2National
                                                                   m



                     Biotechnologies, Sapienza University of Rome;                       Research Council of Italy (SCITEC-CNR); 3Deptof
                     3Mediterranea Cardiocentro-Napoli; 4Department of                   Pharmaceutical Sciences, Università degli Studi di
                                                      om




                     General Surgery and Surgical Speciality Paride                      Milano, Italy
                     Stefanini, Sapienza University of Rome; 5Laboratory
                     affiliated with the Institute Pasteur Italy – Cenci                 Background and Aims: Coronavirus-2 (SARS-CoV-2)
                                                                                         infection causes a sustained prothrombotic state driven
                                                   -c




                     Bolognetti Foundation, Department of Drug Chemistry
                     and Technologies, Sapienza University of Rome; 6IRCCS               by a massive Tissue Factor (TF) expression in circulat-
                                          on




                     Neuromed; 7Department of Public Health and Infectious               ing platelets, leukocytes and microvesicles. Whether
                     Diseases, Sapienza University of Rome; 8Department of               also circulating small extracellular vesicles (sEVs), in
                     Molecular Medicine, Laboratory of Virology, Sapienza                addition to large microvesicles (MVs), can contribute to
                                     N




                     University of Rome; 9Department of Radiological,                    this hypercoagulable scenario through TF expression is
                     Oncological and Pathological Sciences, Sapienza                     not yet known, mainly due to methodological issues in
                     University of Rome, Italy                                           detecting and sizing the smallest vesicles. Therefore,
                                                                                         aim of this study was to characterize TF expression and
                     Background and Aims: Severe acute respiratory syn-                  activity in circulating sEVs, compared to that of MVs,
                     drome coronavirus 2 (SARS-CoV-2) is associated with                 of COVID-19 patients during acute phase infection and
                     an increased risk of venous and arterial thrombosis but             after symptom remission. Methods: MVs and sEVs
                     the underlying mechanism is still unclear. Methods: We              were isolated by plasma differential centrifugation from
                     performed a cross-sectional analysis of platelet function           10 COVID-19 patients enrolled at acute phase infection
                     in 25 SARS-CoV-2 and 10 healthy subjects (HS) by                    (T0) and at six-month-follow-up (T1). Ten healthy sub-
                     measuring Nox2-derived oxidative stress and throm-                  jects (HS) were analyzed as controls. sEVs were count-
                     boxane (Tx) B2 and investigated if administration of                ed by Nano Tracking Assay. In sEVs TF expression was
                     monoclonal antibodies against the Spike (S) protein of              analyzed within CD9/CD63/CD81pos events by imaging
                     SARS-CoV-2 affects platelet activation. Furthermore,                flow cytometry (IFC) and ExoView™ microarray,
                     we investigated in vitro if the S protein of SARS-CoV-              while TF activity by FXa generation assay. TF expres-
                     2 or plasma from SARS-CoV-2 enhanced platelet acti-                 sion and activity in MVs were evaluated for compari-
                     vation (PA). Results: Ex vivo studies showed enhanced               son. Results: By IFC analysis COVID-19 patients at T0
                     platelet Nox2-derived oxidative stress and TxB2                     have about 1.5- and 4-fold higher number of TFpos-sEVs




         [page 12]                                              [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 13



                                                                       XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    and -MVs, respectively, compared to HS, with a trend       Samples were stained using the following antibodies:
                    toward reduction to physiological levels at T1. By         anti-CD62P-selectin-PE and anti-Annexin-V-APC as
                    microarray analysis sEVs behaved similarly (36±12 and      platelet activation markers and anti-CD41-PC7 as spe-
                    25±10 TFpos-spots at T0 and T1, respectively;              cific platelet glycoprotein. The same samples were
                    p=0.0281). sEVs-associated TF is functionally active       stained with anti-IgG-FITC and anti-PF4 and analysed
                    thus able to partially support FXa formation as sEVs       with the ImageStream image cytometer. Patient tissue
                    preincubation with TF-neutralizing antibody reduced        samples, lung and brain, were analyzed by confocal
                    FXa generation by about 30%. However, although sEVs        microscopy for the presence of PF4-IgG complexes.
                    number is significantly higher compared to that meas-      Results: Platelet activation was determined by the
                    ured for MVs (~600-fold in HS), functional activity of     expression of membrane α-granule glycoprotein P-
                    sEV is one-third lower compared to that of MVs.            selectin (CD62P+) and by the presence of phos-
                    Conclusions: These data suggest that, in COVID-19          phatidylserine phospholipids (Annexin-V+). The serum
                    patients, the altered procoagulant phenotype could also    from the patient with VITT was able to activate the
                    be supported by TF carried by sEVs, although their         donor platelets demonstrated by the high expression of
                    functional activity is significantly lower than that of    CD62P+/Annexin-V+. In PRP from healthy subjects, in
                    MVs.                                                       the presence of VITT patient serum and low doses of
                                                                               heparin, the CD62P+/Annexin-V+ expression was
                                                                               decreased and at high doses of heparin, the signal was
                    OC016                                                      further reduced. The ImageStream cytometer demon-




                                                                                                        ly
                                                                               strated the presence of IgG-PF4 complexes on the




                                                                                                on
                                                                               platelet surface. Furthermore, the same complexes were
                    FLOW CYTOMETRY AND MICROSCOPY ASSAY TO
                                                                               observed by confocal microscopy in the patient tissues.
                    IDENTIFY PLATELET ACTIVATING ANTIBODIES
                                                                               Conclusions: The flow cytometry results confirmed the
                    AFTER CHADOX1 NCOV-19 VACCINE                              presence of platelet activating antibodies and that the




                                                                                          e
                                                                               mechanism of action is similar to that observed in
                    C.M. Radu, C. Bulato, E. Campello, S. Gavasso,
                    L. Spiezia, D. Tormene, P. Simioni                             us
                                                                               patients with HIT. Flow cytometry could be a useful
                                                                               assay to identify the real incidence of VITT due to the
                                                                               presence of platelet activating antibodies after vaccina-
                    Department of Medicine, Thrombotic and Hemorrhagic
                                                                               al
                                                                               tion with ChAdOx1 nCOV-19. In addition,
                    Diseases Unit, University of Padua, Italy                  ImageStream cytometry demonstrated the presence of
                                                                       ci

                                                                               IgG-PF4 complexes on the platelet surface. Tissues
                    Introduction: Vaccination against severe acute respira-    analyses demonstrate the presence of IgG-PF4 com-
                                                                 er


                    tory syndrome coronavirus-2 (SARS-Cov-2) is the            plexes. The exact mechanism by which vaccination
                    most important countermeasure to fight the ongoing         with ChAdOx1 nCOV-19 is associated with these
                                                            m



                    Covid-19 pandemic. The vaccine ChAdOx1 nCov-19             platelet-activating antibodies is still unknown.
                    (AstraZeneca) is a recombinant adenoviral vector
                                                om




                    encoding for the spike glycoprotein of SARS-Cov-2.
                    Vaccine-induced thrombotic thrombocytopenia (VITT)
                                                                               OC017
                    is a fatal coagulation disorder after ChAdOx1 nCov-19
                    vaccination, characterized by deep venous thrombosis,
                                             -c




                    pulmonary embolism and cerebral venous sinus throm-        PLATELET-DERIVED EXTRACELLULAR VESICLES
                                                                               CAUSE NEUTROPHIL ACTIVATION IN SYSTEMIC
                                      on




                    bosis in combination with thrombocytopenia. The cause
                    of VITT is due to platelet activating antibodies against   SCLEROSIS
                    platelet factor 4 (PF4) potentially complexed with an
                                 N




                    unknown co-factor making this protein immunogenic.         N. Maugeri, G.A. Ramirez, A. Monno, A. Capobianco,
                    These features make VITT to clinically mimics autoim-      S. Franchini, M.E. Bianchi, P. Rovere-Querini,
                    mune heparin-induced thrombocytopenia (HIT). Aims:         A.A. Manfredi
                    To evaluate by CytoFLEX SRT, ImageStream flow
                    cytometers and confocal microscopy the presence of         San Raffaele Scientific Institute and Vita-Salute San
                    pathogenic antibodies directed against platelets in a      Raffaele University, Italy
                    patient with fatal VITT and cerebral venous thrombosis
                    after adenovirus vaccine ChAdOx1 nCOV-19.                  Background and Aims: Platelet extracellular vesicles
                    Methods: The laboratory results are show a patient         (Plt-EVs) expressing the damage-associated molecular
                    platelet count of 20.000 per cubic millimeter and the      pattern prototypic signal, High Mobility Group Box 1
                    presence of anti-PF4 antibodies detected by enzyme-        (HMGB1) accumulate in the blood of patients with sys-
                    linked immunosorbent assay. The patient had not previ-     temic sclerosis (SSc) and contribute to autoimmunity,
                    ously been exposed to heparin. We analyzed the pres-       fibrosis and vascular inflammation. A poorly understood
                    ence of platelet activating antibodies by CytoFLEX         defect in critical players in sprouting angiogenesis,
                    SRT flow cytometry. Fresh platelet rich plasma (PRP)       angiopoietins and the angiopoietin receptor, Tie2 is also an
                    from healthy donors was incubated with VITT patient        hallmark of SSc. Neutrophils respond to angiopoietins and
                    serum without and in the presence of low and high          other stimuli by undergoing activation and upregulating
                    doses of heparin to inhibit PF4-dependet reaction.         the angiopoietin2, Tie2 expression. Methods: Human




                                                             [Hematology Reports 2021; 13:s3]                                                 [page 13]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 14



         Oral communications




                     studies: We analyzed Tie2 expression by neutrophils in of      and Pathological Sciences, Sapienza, University of Rome,
                     39 patients with SSc and 43 sex- age- matched healthy          Rome, Italy; 5Interventional Cardiology Unit, Santa Maria
                     donors (HD) by flow cytometry. Tie2 expression was con-        Hospital, Terni, Italy6Department of Experimental and
                     firmed by immuno-gold electron microscopy in purified          Clinical Medicine, University of Florence, Firenze, Italy;
                     neutrophils of 20 patients, In vitro experiments: purified     7Stroke Unit, Emergency Department, Sapienza University

                     neutrophils were purified and challenged with recombi-         of Rome, Italy; 8Mediterranea Cardiocentro-Napoli,
                     nant HMGB, or plt-EVs from the plasma of patients with         Naples, Italy
                     SSc. Animal model: Plt-EV were purified from the plasma
                     of patients or HD and injected in the tail vein of NSG         Background and Aims: Vaccine-induced immune
                     mice. After 18 hours blood samples were analyzed by            thrombocytopenia and thrombosis (VITT) is a new syn-
                     flow cytometry and histology of lungs assessed.                drome associated with the ChAdOx1 nCoV-19 adenoviral
                     Neutrophil ablation was induced by treating mice with          vector vaccine against severe acute respiratory syndrome
                     liposome-encapsulated clodronate before Plt-EV injec-          coronavirus 2. Like heparin-induced thrombocytopenia
                     tion. The HMGB1 competitive antagonist, Box A and low          (HIT), this condition has been associated with the devel-
                     molecular weight heparin (LMWH) were used as                   opment of anti-platelet factor 4 antibodies, leading to the
                     inhibitors of HMGB1 in in vitro and in vivo experiments.       neutrophil-platelet aggregate formation and neutrophil
                     Results: Most neutrophils in SSc blood expressed Tie2          extracellular traps (NETs) release. However, the precise
                     (84.7±2.4% vs 8.3±1.1% in HD; p <0.0001), regardless of        molecular mechanisms governing its occurrence are yet to
                     the extent of fibrosis or vascular or systemic inflamma-       be established. Methods: We studied patients with




                                                                                                             ly
                     tion. HMGB1 is a putative candidate signal supporting          ischemic stroke after ChAdOx1 nCoV-19 vaccination,
                                                                                    patients with ischemic stroke from other causes (IS) and




                                                                                                     on
                     neutrophils Tie2 overexpression, as it plays a critical role
                     in neutrophil activation and is present as an EV-associated    healthy subjects (HS). We analyzed NETs release and
                     bioactive molecule in the blood of SSc patients. Our data      coagulation by measuring in plasma and cerebral thrombi
                     support such a role, as HMGB1 and HMGB1+-Plt-EV                citrullination of histone H3 (CitH3) and tissue factor (TF),




                                                                                               e
                     purified from the plasma of patients with SSc induced up-      respectively. Furthermore, we investigated in vitro if plas-
                     regulation of Tie2 in vitro and the event was abrogated by
                     HMGB1 antagonists, Box A and LMWH. Furthermore,
                     18 hours after intravenous infusion of EV purified from
                                                                                        us
                                                                                    ma post-ChAdOx nCoV-19 vaccination enhanced NETs
                                                                                    release. Results: In vivo studies showed enhanced circu-
                                                                                    lating levels of CitH3 and TF in patients with ischemic
                                                                                    al
                     the plasma of SSc patients (but not from the plasma of         stroke after ChAdOx1 nCoV-19 vaccination compared to
                     healthy donors) the fraction of Tie2-expressing neu-           IS and HS groups. In vitro study showed that plasma from
                                                                            ci

                     trophils in the blood increased dramatically. BoxA and         post-ChAdOx nCoV-19 vaccination increased NETs
                     LMWH prevented the neutrophil Tie2 upregulation (>85           release and Protein Arginine Deiminase 4 (PAD4) expres-
                                                                      er


                     and 95% respectively). Histological analysis revealed          sion by neutrophils; this effect was more evident in the
                     fibrosis associated with massive infiltration of the lung by   presence of platelets and blunted by PAD4 inhibitor,
                                                                m



                     Tie2+ neutrophils. Neutrophil ablation by liposome-            heparin, and anti-FCγRII antibody. Finally, immunohisto-
                     encapsulated clodronate prevented SSc Plt-EV-induced           chemistry analysis showed a much stronger staining
                                                   om




                     lung fibrosis, thus causally linking neutrophil activation,    intensity for the antibody against PAD4 in a cerebral
                     Tie2 neutrophil-expressing pulmonary infiltration and tis-     thrombus from a patient after ChAdOx1 nCoV-19 vacci-
                     sue fibrosis. Conclusions: Particulate HMGB1 expressed         nation compared to that from IS patient. Conclusions:
                                                -c




                     on Plt-EVs could be a player and a pharmacological target      This study provides evidence that in VITT patients PAD4
                     in SSc.                                                        overactivation is responsible for NETs release at the level
                                        on




                                                                                    of thrombus growth, thereby its inhibition may represent
                                                                                    a novel approach to the treatment of VITT.
                     OC018
                                   N




                                                                                    OC019
                     NEUTROPHIL ACTIVATION AND NETS RELEASE
                     DRIVER THROMBOSIS IN VACCINE-INDUCED
                     IMMUNE THROMBOTIC THROMBOCYTOPENIA                             A GLOBAL SCORE OF MODIFIABLE
                                                                                    CARDIOVASCULAR RISK FACTORS IN
                     C. Nocella1, S. Bartimoccia2, V. Cammisotto1,                  INDIVIDUALS FREE FROM CARDIOVASCULAR
                     V. Castellani3, M. Leopizzi4, G. D’Amati4,                     DISEASE: LONGITUDINAL FINDINGS FROM THE
                     M. Dominici5, R. Marcucci6, M. De Michele7,                    MOLI-SANI STUDY
                     R. Carnevale2,8, P. Pignatelli1,8, F. Violi8
                                                                                    M. Persichillo1, M. Bonaccio1, S. Costanzo1,
                     1Department  of Clinical Internal, Anaesthesiological and      A. De Curtis1, S. Magnacca3, F. Bracone1, T. Panzera1,
                     Cardiovascular Sciences, Sapienza University of Rome,          C. Cerletti1, M.B. Donati1, G. de Gaetano1,
                     Rome, Italy; 2Department of Medical-Surgical Sciences          L. Iacoviello1,2, A. Di Castelnuovo3 on behalf of the
                     and Biotechnologies, Sapienza University of Rome, Latina,      Moli-sani Study Investigators
                     Italy; 3Department of General Surgery and Surgical
                     Speciality Paride Stefanini, Sapienza University of Rome,      1Department
                                                                                            of Epidemiology and Prevention, IRCCS
                     Rome, Italy; 4Department of Radiological, Oncological          NEUROMED, Pozzilli (IS); 2EPIMED Research Center,




         [page 14]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 15



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Department of Medicine and Surgery, University of              design and evaluate the efficacy of a preventive
                    Insubria, Varese; 3Mediterranea Cardiocentro, Napoli,          approach, to compare different strategies and to com-
                    Italy                                                          municate effectiveness of a preventive action, thus
                                                                                   increasing compliance to the strategy itself.
                    Background and Aims: Cardiovascular (CV) disease,
                    the leading cause of death globally, is largely preventa-      Table 1.
                    ble through targeting modifiable risk factors. Scores of
                    modifiable CV risk factors able to collect the combined
                    effect of all the included components were mainly
                    developed in US populations, or do not include nutri-
                    tional aspects. Beside the effect of specific prevention
                    strategies, the CV risk profile based on modifiable CV
                    risk factors may change in time, but data dealing with
                    this variation over long periods are scarce. Based on
                    data from an Italian population, we aimed a) to compute
                    an algorithm based on common modifiable CV risk fac-
                    tors, including diet and b) to assess the variation at indi-
                                                                                   OC020
                    vidual level of 9 common modifiable CV risk factors,
                    and of the global score based on them, after 12.7 years




                                                                                                           ly
                    of follow-up. Methods: We analyzed data on 1,873 par-          HIGH, BUT NOT LOW-, DOSE ATORVASTATIN
                    ticipants from a sub-cohort of the larger Moli-sani            PREVENTS THE INCREASE OF CIRCULATING




                                                                                                    on
                    Study (enrolled 2005-2010) that was re-examined in             MMP-2 LEVELS IN PATIENTS WITH NSTE-ACUTE
                    2017-2020. Individuals were randomly recruited from            CORONARY SYNDROMES
                    those in the original cohort with longer follow-up and




                                                                                              e
                    were free from CV disease. The panel of modifiable CV          F. Paciullo1, E. Falcinelli1, M. Del Pinto2, T. Fierro1,
                    risk factors included smoking habit, adhesion to
                    Mediterranean diet, LDL, HDL, triglycerides, mean
                    arterial pressure, glucose, relative fat mass (proxy of
                                                                                       us
                                                                                   P. Gresele1
                                                                                   1Department     of Medicine and Surgery, Section of
                                                                                   al
                    body fat percentage) and leisure-time physical activity.       Internal and Cardiovascular Medicine, University of
                    A global score of modifiable CV risk factors was calcu-        Perugia, Italy; 2Division of Cardiology, Perugia
                                                                           ci

                    lated as a weighted sum of the nine standardised risk          Hospital, Italy
                    factors. Weights reflect the strength of the risk
                                                                    er


                    factor/outcome association. To derive the algorithm, we        Background and Aims: High-dose statins improve car-
                    used data on 16,656 men and women (aged ≥45 years)             diovascular outcomes in patients with acute coronary
                                                               m



                    from the population of the Moli-sani Study free from           syndromes (ACS). The beneficial effect of statins are
                    CV disease, and tested the combined association of the         considered to be largely due to their lipid lowering action
                                                  om




                    9 risk factors with a composite outcome of fatal and           but, especially upon very short term administration, they
                    non-fatal CV events. Results: In multivariable Cox             may also be the consequence of pleiotropic actions.
                    regression analysis adjusted for non-modifiable risk           Metalloproteinases (MMP), a family of enzymes degrad-
                                               -c




                    factors, one unit more in the score was associated with        ing extracellular matrix, are released in blood during
                    a hazard ratio for CV events equal to 1.07 (95%CI: 1.06        acute coronary syndromes and have been reported to
                                       on




                    to 1.08). The re-analysed sample consisted of 1,873            contribute to atheroma unstabilization and recurrent car-
                    individuals, 55% women, 57 (SD=7) years of age. In             diovascular events. In particular, it has been recently
                    12.7 median years of follow-up, an improvement in the          reported that in patients with ACS high MMP-2 levels
                                  N




                    CV risk profile was observed as number of cigarettes,          are associated with the degree of myocardial injury,
                    mean arterial pressure and LDL were reduced, and HDL           measured as high-sensitivity troponin T levels, and can
                    increased (Table 1). On the contrary, a worsening in the       predict the severity of outcome. Moreover, in a cohort of
                    CV risk profile was detected as adherence to                   patients with AMI MMP-2 levels predicted all-cause
                    Mediterranean diet was reduced and relative fat mass           mortality. In the present study we investigated the effect
                    and glucose was increased (Table 1). Mean values of            of atorvastatin at high vs standard doses on the early
                    triglycerides and physical activity remained unchanged         increase of circulating MMP-2 levels in patients with
                    (Table 1). Because of these variations, the global score       NSTE-acute coronary syndrome (ACS). Methods: In
                    of modifiable CV risk factors diminished of nearly 1           this single center study we prospectively collected clini-
                    point (Table 1). Conclusions: In the last 13 years, lipid      cal and laboratory data from patients with a definite diag-
                    profile and smoking habit of a large Italian population,       nosis of NSTE-ACS. Patients were divided in three
                    originally free from CV disease, improved, whereas             groups according to the statin regimen: atorvastatin 80
                    adherence to Mediterranean diet, fat distribution and          mg/die from day 1 (high-dose group); atorvastatin 20 mg
                    glycaemic control worsened. These findings can help            from day 1 (low-dose group); atorvastatin 20 mg starting
                    setting strategies for cardiovascular prevention. We pro-      later than 96 hours after admission (non statin group).
                    pose an algorithm based on nine modifiable CV risk             Dose regimens were chosen by the caring cardiologist
                    factors, easily to replicate and predictive of fatal and       based on LDL levels at hospital admission. Each patient
                    non-fatal CV events. The score can be managed to               was clinically evaluated on admission and after 3, 5 and




                                                                [Hematology Reports 2021; 13:s3]                                                 [page 15]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 16



         Oral communications




                     30 days from the event, and blood samples for the meas-       gle-center hospital-based study enrolling patients with
                     urement of MMP-2 were collected. Peripheral blood was         atrial fibrillation, aged 65 years or older, and with no con-
                     collected in parallel also from a group of age- and sex-      traindications to undergo magnetic resonance imaging,
                     matched healthy controls (HC). Plasma MMP-2 levels            referring to Center of Atherothrombotic Disease of our
                     were measured by zymography. Serologic, clinical and          University Hospital (AOU Careggi) for the management
                     instrumental parameters were compared in three groups.        of oral anticoagulation therapy. Recruited patients are
                     Data are reported as mean±SEM. Results: 40 consecu-           evaluated by means of a comprehensive protocol, with
                     tive patients were enrolled, 13 patients in the high dose     clinical, cerebral magnetic resonance imaging and circu-
                     group, 14 in the low-dose group and 13 in the non statin      lating biomarkers assessment. The main outcomes are
                     group. Characteristics of patients in the three groups        the evaluation of cerebral microangiopathy using MRI
                     were comparable (age: 67±2, 70±3, 69±3; sex (F): 38%,         (lacunar infarcts, non-lacunar infarcts, microbleeds,
                     21%, 15%; prevalence of risk factors: 70%, 50%, 50%;          hyperintensity of the white matter, SVDs [small vessel
                     hours from symptom onset: 20±8, 24±6, 11±7, respec-           disease score]) and the onset of ischemic or hemorrhagic
                     tively) Peripheral blood plasma MMP-2 levels at enroll-       stroke related to the levels of circulating biomarkers of
                     ment were significantly higher in NSTE-ACS patients           inflammation (IL-6, IL-8, TNFα, IL-4, IL-10, CCL2,
                     than in HC (median 918±72 ng/ml vs 682±97 ng/ml,              CXCL10, ICAM-1, VCAM-1, VEGF), haemostasis
                     p<0.01) but did not differ significantly between the three    (PAI-1, CLT, vWF) and extracellular matrix remodeling
                     patient groups. High-dose atorvastatin significantly          (MMP-2,-7,-8,-9,-12, TIMP-1,-2,-3,-4). Starting from
                     reduced plasma MMP-2 levels at 3 days compared to             September 2017, 191 patients (mean age 78.1±6.7, range




                                                                                                            ly
                     other strategies (High-dose=before: 986.4±90ng/ml             65-97; 65.3% males) were enrolled. 56 patients (29.3 %)
                     day3:781±81ng/ml; Low-dose=before: 842.4±148ng/ml




                                                                                                    on
                                                                                   were on vitamin K antagonists, and 135 (70.7 %) were on
                     day3:895.2±120 and non statin=before:930.2±1259               direct oral anticoagulants. Follow-up clinical evaluation
                     day3: 1033±123ng/ml, p<0.05, respectively, for high-          and brain MRI were performed at 18 months. Results: At
                     dose vs low-dose and vs non statin regimen) (Figure 1).       multivariate analysis, adjusted for age, sex, CHA2DS2-




                                                                                              e
                     Conclusions: High-dose, but not low-dose, statin              VASc, HAS-BLED and type of anticoagulant, independ-
                     reduces the increase of circulating MMP-2 triggered by
                     ACS. Given that high circulating MMP-2 levels in
                     patients with ACS can predict poor prognosis and are
                                                                                       us
                                                                                   ent predictors were: low levels of vWF and elevated lev-
                                                                                   els of TIMP-2 for microbleeds [OR: 0.62 95% CI (0.42-
                                                                                   0.89), p=0.011 and OR: 1.25 (1.01-1.77), p=0.049
                                                                                   al
                     associated with increased incidence of cardiovascular         respectively]; elevated levels of IL-6 and TIMP-4 for
                     events, this reduction may contribute to explain the pro-     hyperintensity of white matter [OR: 1.55 (1.01-2.58),
                                                                           ci

                     tective effect of high dose statin after ACS on recurrent     p=0.049 and OR: 1.44 (1.01-2.07), p=0.047 respective-
                     cardiovascular events.                                        ly]; elevated levels of MMP-2 and of TIMP-1 and TIMP-
                                                                     er


                                                                                   2 for lacunar infarcts [OR: 1.33 (1.01-1.89), p=0.049,
                                                                                   OR: 1.48(1.01-2.18), p=0.048 and 1.50 (1.05-2.16),
                                                               m



                     OC021                                                         p=0.028 respectively]; elevated levels of TIMP-1, TIMP-
                                                                                   2, TIMP-3 for the appearance of at least 1 new lesion
                                                   om




                     INFLAMMATION, HAEMOSTASIS AND                                 according to the SVD score [OR: 1.48 (1.01-2.19),
                     EXTRACELLULAR MATRIX IN THE                                   p=0.046, OR: 1.53 (1.05-2.23), p=0.028 and OR: 1.58
                                                                                   (1.09-2.31), p=0.016 respectively]. Conclusions: The
                     STRATIFICATION OF BLEEDING AND
                                                -c




                                                                                   Strat-AF study may be an essential step towards the
                     THROMBOTIC RISK OF ATRIAL FIBRILLATION ON                     exploration of the role of a combined clinical biomarker
                                        on




                     ORAL ANTICOAGULANT THERAPY: INSIGHTS                          or multiple biomarker models in predicting stroke risk in
                     FROM STRAT-AF STUDY                                           atrial fibrillation and might sustain the incorporation of
                                                                                   such new markers in the existing stroke prediction
                                   N




                     F. Alfano1, A.M. Gori1, F. Cesari1, E. Salvadori2,            schemes by the demonstration of a greater incremental
                     M. Berteotti1, B. Giusti1, A. Kura1, G. Cassioli1,            value in predicting stroke risk and improvement in clini-
                     E. Lotti1, D. Poli1, A. Poggesi2, R. Marcucci1                cal outcomes in a cost-effective fashion.
                     1Department   of Experimental and Clinical Medicine,
                     University of Florence, Florence, Italy; 2NEUROFARBA          OC022
                     Department, Neuroscience Section, University of
                     Florence, Florence, Italy
                                                                                   CIRCULATING BIOMARKERS AND BLOOD-BRAIN-
                     Background: In anticoagulated atrial fibrillation             BARRIER LEAKAGE: RESULTS FROM THE
                     patients, the validity of models recommended for the          REPERFUSION INJURY IN ISCHEMIC STROKE
                     stratification of the risk ratio between benefits and hem-    (RISK) STUDY
                     orrhage risk is limited. We hypothesize that biological
                     markers – both circulating and neuroimaging-based –           A. Bertelli1, A. Kura1, B. Piccardi2, F. Arba2,
                     and their possible interaction, might improve the predic-     G. Barbieri1, R. De Cario1, A. Sodero2, B. Giusti1,
                     tion of bleeding risk in atrial fibrillation patients under   A.M. Gori1, R. Marcucci1, P. Nencini2
                     treatment with any type of oral anticoagulant. Methods:
                     The Strat-AF study is an observational, prospective, sin-     1Dept of Experimental and Clinical Medicine, University




         [page 16]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 17



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    of Florence, Florence, Italy; 2NEUROFARBA                     /anti-inflammatory ratios were associated with poor out-
                    Department, Neuroscience Section, University of               come assessed by three-month mRS. Our results support
                    Florence, Italy                                               a possible link between BBB disruption and consequent
                                                                                  inflammatory cascade in acute ischemic stroke and sug-
                    Aim: To evaluate the effect of circulating inflammatory       gests the pathogenetic role of inflammation into 3 month
                    mediators and matrix metalloproteinases (MMP) on              neurological disability.
                    blood-brain-barrier (BBB) leakage in the acute phase of
                    stroke. Methods: Observational prospective single-cen-
                    tre hospital-based study that included consecutive acute      OC023
                    ischaemic stroke patients treated with intravenous throm-
                    bolysis, endovascular treatment or both. Circulating bio-
                                                                                  ENHANCED PLATELET REACTIVITY IN
                    markers were taken before and after 24 hours from acute
                    interventions. We assessed pre-treatment BBB leakage          GESTATIONAL DIABETES
                    with CT perfusion by using Ktrans within the ischaemic
                    area. BBB disruption was evaluated with Ktrans maps           G. Guglielmini1, E. Falcinelli1, E. Piselli1,
                    automatically provided by Olea Sphere within the              A.M. Mezzasoma1, L. Alfonsi2, C. De Luca2, V. Fino2,
                    ischemic core and the ischemic penumbra. We evaluated         E. Torlone3, P. Gresele1*, S. Gerli2*
                    independent associations between BBB leakage and bio-         1Division
                    markers levels at baseline, 24 hours and their relative                  of Internal and Cardiovascular Medicine;




                                                                                                         ly
                                                                                  2Division  of Obstetrics and Gynecology; 3Division of
                    variations [(Δ=24hours level-baseline level)/baseline
                    level] adjusting for age, sex, baseline stroke severity,      Endocrinology and Metabolism; Department of




                                                                                                  on
                    type of treatment and onset to treatment time. Results:       Medicine and Surgery, University of Perugia, Perugia,
                    We included 100 patients, mean (±SD) age was                  Italy; *these authors share senior authorship
                    75.9±11.6, 51 (51%) males, median (IQR) NIHSS was




                                                                                              e
                    17 (10-22). Endovascular treatment alone was performed        Background and Aims: Gestational diabetes mellitus
                    in 47 (47%) patients, intravenous thrombolysis in 25
                    (25%) patients, and 28 patients (28%) received both
                    treatments. Baseline biomarker levels were not signifi-
                                                                                     us
                                                                                  (GDM) is a disorder of carbohydrate metabolism which
                                                                                  occurs in around 14% of pregnancies. While platelet
                                                                                  hyperresponsiveness in type-2 diabetes mellitus
                                                                                  (T2DM) has been well characterized and shown to play
                                                                                  al
                    cantly different among the 3 types of treatment. We did
                    not find any association between baseline biomarker val-      a crucial role in cardiovascular complications, this
                                                                          ci

                    ues and BBB leakage. BBB leakage was correlated with          aspect has been little studied in GDM. Aim of our study
                    24h MMP-2 (Rho 0.22, p=0.045), 24h IL-12 (Rho 0.26,           was to evaluate platelet activation in vivo, ex vivo
                                                                    er


                    p=0.017), and with ΔIL-12 levels (Rho 0.22; p=0.047).         platelet hyperreactivity and endothelial function in
                    When we dichotomized BBB values according to the              GDM compared with normal pregnancy. Methods: For
                                                              m



                    mean for the whole population (“low” BBB disruption           the GDM group inclusion criteria were glucose intoler-
                    with values up to 0.63; “high” BBB disruption with val-       ance confirmed by a pathologic oral glucose tolerance
                                                  om




                    ues higher than 0.63), patients with higher BBB had sig-      test performed between 16 and 18 weeks of gestation.
                    nificantly higher levels of 24h MMP-7 (p=0.010), 24h          For the control pregnant (CP) group the only inclusion
                    MMP-9 (p=0.019) and 24 h IL-12 (p=0.010). At multi-           criterion was a physiological pregnancy. Groups were
                                                                                  balanced for age and gestational age. Exclusion criteria
                                               -c




                    variate logistic regression analysis, adjusted for the
                    above mentioned variables, higher BBB leakage was sig-        were: pregestational T2DM, hypertension, preeclamp-
                                                                                  sia, BMI>30, pregestational hypothyroidism, hemor-
                                       on




                    nificantly associated with 24h IL-12 levels [OR=1.81 (CI
                    1.06-3.08), p=0.030], 24h MMP-7 [OR=2.80 (CI 1.28-            rhagic diathesis, platelet abnormalities, platelet count
                    6.14), p=0.010] and 24h MMP-9 [OR=1.89 (CI 1.17-              <100,000 cells/µl, or thrombotic events within 6
                                  N




                    3.04), p=0.009]. Modified Rankin disability score (mRS)       months. All tests were conducted at 26-28 weeks of ges-
                    was dichotomized into good (mRS 0–2) (n=45) or poor           tation (T1), at 34-36 weeks (T2) and 8 weeks post-par-
                    (mRS 3–6) (n=55) outcome. Patients with poor outcome          tum (T3). In vitro: collagen-, ADP-, Arachidonic Acid
                    had significantly higher levels of baseline IL6, CCL2 and     (AA)-, epinephrine- U46619- and TRAP-6-induced
                    CXCL10 [IL6:199 (131-350) pg/mL vs 145 (112-206)              LTA; PFA-100® closure time, platelet activation anti-
                    pg/mL, p=0.0401; CCL2: 10.74 (0.48-1.27) ng/mL vs             gens and platelet production of reactive oxygen species
                    0.52 (0.35-0.76) ng/mL, p=0.005 and CXCL10: 1.4 (1.1-         (ROS) and nitric oxide (NO) upon in vitro stimulation.
                    2.3) ng/mL vs 0.9 (0.6-1.6) ng/mL, p 0.025] than patients     In vivo: Endothelial function measured using the
                    with mRS 0-2. IL6/IL4 and IL6/IL10 ratios were signif-        EndoPAT™ System. Results: 46 pregnant women, 23
                    icantly higher in patients with poor outcome [1.28 (0.91-     GDM and 23 CP, were enrolled. Low dose collagen-,
                    2.33) vs 0.86 (0.59-1.10), p<0.001; 0.74 (0.48-1.27) vs       ADP- and AA- induced platelet aggregation increased
                    0.52 (0.34-0.76), p=0.020] than in patients with mRS 0-       significantly from T1 to T2, both in CP and GDM
                    2. At multivariate logistic regression analysis, the ratios   women, to decrease then at T3. However, GDM showed
                    IL6/IL4 and IL6/IL10 were significantly associated with       a significantly higher LTA at T2 compared with CP
                    three-month mRS 3–6 [OR: 2.48 (1.01-10.11), p=0.049           which persisted at T3. EC50 for collagen LTA was sig-
                    and OR:2.99(1.05-8.54), p=0.040 respectively].                nificantly lower in GDM. Epinephrine- U46619- and
                    Conclusions: Early BBB leakage was associated with 24         TRAP-6-induced platelet aggregation increased signifi-
                    hours IL12, MMP-7 and MMP-9 levels. Unbalanced pro-           cantly throughout pregnancy in both CP and GDM




                                                               [Hematology Reports 2021; 13:s3]                                              [page 17]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 18



         Oral communications




                     women, but then decreased at T3 in CP, while GDM              with ABI measurement. We used an optimized cut-off
                     women maintained a significantly higher platelet reac-        of PCSK9 >1150 pg/ml obtained from ROC curve
                     tivity. PFA-100 C-EPI closure time was significantly          analysis. All-cause mortality according to the ABI value
                     shorter in GDM than in CP at all the observation time         was also analysed. Results: 115 (19.9%) had an ABI
                     points. Platelet adhesion to a collagen-coated surface        ≥1.4. The mean age was 72.1 years and 42.1% of
                     under high-flow was significantly higher in GDM than          patients were women. Patients with ABI ≥1.4 were
                     in CP. The expression of P-Sel or PAC-1 upon stimula-         older, more frequently male and diabetic. Multivariable
                     tion with ADP or TRAP was significantly higher in             logistic regression analysis showed an association
                     GDM compared with CP at T1 and T2. Platelet sensitiv-         between ABI ≥1.4 and serum levels of PCSK9 >1150
                     ity to the inhibitory effect of NO was significantly lower    pg/ml (Odds Ratio 1.835, 95%CI 1.133-2.970,
                     in GDM compared to CP. Also, platelet NO production           p=0.014). During a median follow up of 41 months, 112
                     was significantly lower in GDM than in CP women at            deaths occurred. At multivariable Cox regression analy-
                     T1 and further decreased at T2. Arterial stiffness by the     sis, ABI ≥1.4 was associated with an increased risk of
                     EndoPAT™ System was significantly higher in GDM at            mortality (Hazard Ratio 1.676, 95%CI 1.073-2.617,
                     all time points. Conclusions: Our data show that GDM          p=0.023). Conclusions: In AF patients, PCSK9 levels
                     generates a condition of platelet hyperreactivity. A par-     relate to an abnormally high ABI, which is in turn asso-
                     ticularly evident defect is represented by a lower sensi-     ciated with an increased mortality risk. This is the first
                     tivity of platelets to the inhibitory action of NO and by a   clinical evidence of an association between PCSK9 and
                     reduced platelet and endothelial NO production, while         vascular calcification in AF patients




                                                                                                           ly
                     platelet oxidative stress is increased, contributing to the




                                                                                                   on
                     hyperreactivity state of platelets in GDM. In conclusion
                     GDM generates a platelet hyperreactivity condition            OC025
                     which tends to worsen as pregnancy progresses, and per-
                     sists for at least 2 months after delivery. Platelet hyper-




                                                                                              e
                                                                                   HEAD-TO-HEAD COMPARISON OF THE EFFECT OF
                     reactivity may contribute to long term cardiovascular
                     complications in GDM patients.
                                                                                       us
                                                                                   PLATELET- AND MONOCYTE-DERIVED
                                                                                   MICROVESICLES ON ENDOTHELIAL FUNCTIONS

                                                                                   M. Brambilla1, P. Canzano1, M. Talmon2, L. Fresu2,
                                                                                   al
                     OC024
                                                                                   S. Brunelleschi2, E. Tremoli3, M. Camera1,4
                                                                           ci

                     PROPROTEIN CONVERTASE SUBTILISIN/KEXIN                        1Centro Cardiologico Monzino I.R.C.C.S., Milan, Italy;
                                                                     er


                     TYPE 9 (PCSK9) LEVELS AND ABNORMALLY                          2Dept  of Health Sciences, University of Piemonte
                     HIGH ANKLE-BRACHIAL INDEX IN ATRIAL                           Orientale, Novara, Italy; 3Maria Cecilia Hospital,
                                                               m



                     FIBRILLATION                                                  Cotignola, Italy; 4Dept of Pharmacological and
                                                                                   Biomolecular Sciences,Università degli Studi di Milano,
                                                   om




                     P. Pignatelli1,4, D. Menichelli1, G. Galardo2,                Milan, Italy
                     V. Cammisotto1, S. Bartimoccia1, R. Carnevale3,4,
                     D. Pastori1                                                   Background and Aims: Circulating microvesicles
                                                                                   (MVs) are recognized as both biomarkers and media-
                                                -c




                     1Department   of Clinical Internal, Anesthesiological and     tors of inflammation and endothelial dysfunction in
                                        on




                     Cardiovascular Sciences, Sapienza University of Rome,         many pathological conditions, including cardiovascular
                     Rome, Italy; 2Department of General Surgery, Surgical         diseases. The two main MV-subsets mediating these
                     Specialties and Organ Transplantation “Paride                 effects are those derived from monocytes and platelets
                                   N




                     Stefanini,” Sapienza University of Rome, Rome, Italy;         (PLT), although the specific contribution of each of
                     3Department of Medical-Surgical Sciences and                  them is not well defined. Thus, aim of this study was to
                     Biotechnologies, Sapienza University of Rome, Latina,         assess how MVs released from PLT and monocytes
                     Italy; 4Mediterranea Cardiocentro, Naples, Italy              influence processes involved in the vessel damage -i.e.
                                                                                   oxidative stress, inflammation, leukocyte-endothelial
                     Background and Aims: High ankle-brachial index                adhesion. Methods: PLT and monocytes isolated from
                     (ABI) has been associated with increased risk of worse        healthy subjects (HS, n=10) were stimulated with
                     outcomes in the general population. Few data on atrial        TRAP-6 (20 µM) and LPS (10µg/ml) to release MVs
                     fibrillation (AF) do exist. Experimental data suggest         that were isolated, characterized by flow cytometry, and
                     that proprotein Convertase Subtilisin/Kexin type 9            added to the culture medium of human vascular
                     (PCSK9) contribute to vascular calcification but clini-       endothelial cells (hECV). Superoxide anion production,
                     cal data on this association are lacking. To investigate      inflammatory markers (IL6, TNFalpha, NF-κB mRNA
                     the relationship between circulating PCSK9 levels and         expression) and hECV adhesiveness after MVs chal-
                     abnormally high ABI in patients suffering from AF.            lenge were evaluated. MVs spontaneously released
                     Methods: We analysed data from 579 patients included          from cells of acute myocardial infarction patients
                     in the prospective ATHERO-AF study. An ABI ≥1.4               (AMI; n=10) were used to verify whether the in vivo
                     was considered as high. Patients with an ABI <0.9 were        setting reflected the in vitro activation. Results:
                     excluded. PCSK9 levels were measured coincidentally           Incubation of hECV with MVs released by HS-activat-




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 19



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    ed PLT and monocytes triggered an oxidative burst (3-        grade of inflammation not balanced by anti-inflammato-
                    fold increase) in a MV concentration-dependent man-          ry cytokines; patients with ratios in the other tertiles were
                    ner. Monocyte-MVs doubled IL6, TNFalpha, and NF-             assigned as patients with a low level of inflammation.
                    κB mRNA expression and monocyte-endothelial adhe-            Starting from September 2017, 191 patients (mean age
                    sion. These functions were only slightly influenced by       78.1±6.7, range 65-97; 65.3% males) were enrolled. 56
                    PLT-MVs. These effects were associated with an anti-         patients (29.3 %) were on vitamin K antagonists, and 135
                    genic signature, i.e. procoagulant and PLT-Psel+ and         (70.7 %) were on direct oral anticoagulants. Results:
                    monocyte-CD16+ MVs, reflecting the in vitro stimula-         Patients treated with DOACS had higher ratio values
                    tion and also found in AMI MVs. Interestingly howev-         than patients treated with AVK [IL6/IL4: 0.15 (0.07-
                    er, AMI-monocyte-MVs supported adhesion of mono-             0.35) vs 0.05 (0.03-0.25), p=0.001; IL6/IL10: 0.70 (0.28-
                    cytes to hECV as observed with HS-MVs but not the            5.49) vs 0.54 (0.18-1.42), p=0.090; TNF-alpha/IL4: 0.16
                    oxidative stress. Conversely, AMI PLT-MVs sustained          (0.12-0.31) vs 0.13 (0.08-0.31), p=0.055; TNF-
                    not only redox state but also the inflammatory pheno-        apha/IL10: 0.97 (0.25-5.23) vs 0.75 (0.51-2.62),
                    type (two-fold increase). Conclusions: These data pro-       p=0.105]. According to pro-anti-inflammatory ratio ter-
                    vide evidence that MVs derived from activated PLT and        tiles, 28/135 DOAC patients (20.7%) had an elevated
                    monocytes differently affect endothelial behavior.           grade of inflammation not balanced by anti-inflammato-
                    These functional effects are partially shared also by        ry cytokines, whereas in AVK group only 5.4% (3/53)
                    MVs spontaneously released by platelets and mono-            were classified as patients with high levels of inflamma-
                    cytes from AMI patients and support the antiplatelet         tion (p=0.009). At multivariate logistic regression analy-




                                                                                                          ly
                    treatment in this clinical setting.                          sis, adjusted for age, sex, CHA2DS2-VASc, HAS-
                                                                                 BLED, AVK treatment resulted an independent and pro-




                                                                                                  on
                                                                                 tective predictor for having a high grade of inflammation
                    OC026                                                        not balanced by anti-inflammatory cytokine levels
                                                                                 [OR=0.27 (0.08-0.97), p=0.045]. Conclusions: These




                                                                                            e
                                                                                 result from Strat-AF study may be an essential step
                    PLEIOTROPIC EFFECTS OF ANTICOAGULANT
                    THERAPIES: IS THERE A DIFFERENCE BETWEEN
                    AVK AND DOAC?
                                                                                     us
                                                                                 towards the exploration of the role and the contribute of
                                                                                 anticoagulant therapy in reducing inflammation-related
                                                                                 biomarkers in AF patients. Inflammation has also been
                                                                                 al
                                                                                 associated with endothelial dysfunction, coagulation cas-
                    F. Alfano1, A.M. Gori1, F. Cesari1, E. Salvadori2,           cade activation and thrombogenesis. Thus, inflammation
                    B. Bandinelli1, B. Giusti1, A. Bertelli1, A. Kura1,
                                                                          ci

                                                                                 may contribute to both the occurrence/maintenance of
                    E. Lotti1, R. De Cario1, A. Poggesi2, R. Marcucci1           AF and its thromboembolic complications. These results
                                                                   er


                    1Department
                                                                                 might sustain the incorporation of such new markers in
                                  of Experimental and Clinical Medicine,         the existing stroke prediction schemes by the demonstra-
                                                             m



                    University of Florence, Florence, Italy; 2NEUROFARBA         tion of a greater incremental value in predicting stroke
                    Department, Neuroscience Section, University of              risk and improvement in clinical outcomes in a cost-
                                                 om




                    Florence, Florence, Italy                                    effective fashion.
                    Background: Atrial fibrillation (AF) is the most com-
                    mon supraventricular arrhythmia. Emerging evidence
                                              -c




                                                                                 OC027
                    suggests a significant role of inflammation in the patho-
                                       on




                    genesis and in the maintenance of AF. We hypothesize a
                    different role of anticoagulant therapy (AVK vs. DOACs)      IN VITRO CHARACTERIZATION OF
                    in reducing the levels of inflammatory biomarkers in AF.     PATIENT-SPECIFIC ENDOTHELIAL COLONY-
                                  N




                    Methods: The Strat-AF study is an observational,             FORMING CELLS TO INVESTIGATE THE ROLE OF
                    prospective, single-center hospital-based study enrolling    ENDOTHELIAL DYSFUNCTION IN THE
                    patients with atrial fibrillation, aged 65 years or older,   PATHOGENESIS OF UNPROVOKED VENOUS
                    and with no contraindications to undergo magnetic reso-      THROMBOEMBOLISM
                    nance imaging, referring to Center of Atherothrombotic
                    Disease of our University Hospital (AOU Careggi) for         A. Cancellara1, M. Bacci2, L. Bertolani1, R. Ciceri1,
                    the management of oral anticoagulation therapy.              E. Romualdi3,4, V. Pessi4, M. Fantuzzi4, C. Lodigiani2,
                    Recruited patients are evaluated by means of a compre-       M.P. Donadini4,5, S. Della Bella1, F. Calcaterra1*,
                    hensive protocol, with clinical and circulating biomark-     D. Mavilio1*
                    ers assessment. One of the main outcomes is the evalua-
                    tion of the grade of inflammation in patients treated with   1Unit   of Clinical and Experimental Immunology,
                    AVK vs. DOACs. In order to evaluate the grade of             Department of Medical Biotechnologies and
                    inflammation, we calculated the ratios between pro- and      Translational Medicine, University of Milan, Milan,
                    anti-inflammatory cytokines (IL6/IL4, IL6/IL10, TNF-         Italy; Humanitas Clinical and Research Center - IRCCS,
                    alpha/IL4, TNF-alpha/IL10). The pro-/anti-inflammatory       Rozzano, Milan, Italy; 2Center for Thrombosis and
                    ratios were divided into tertiles and were used to calcu-    Haemorragic Diseases, IRCCS Humanitas Research
                    late a score of inflammation. Patients with ratios in the    Hospital, Rozzano, Milan, Italy; 3UO Medicina 2,
                    third tertile were assigned as patients with an elevated     Ospedale di Circolo e Fondazione Macchi, ASST Sette




                                                              [Hematology Reports 2021; 13:s3]                                                   [page 19]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 20



         Oral communications




                     Laghi, Varese, Italy; 4Università dell’Insubria,              OC028
                     Dipartimento di Medicina e Chirurgia, Varese, Italy;
                     5Centro Trombosi ed Emostasi, Ospedale di Circolo e

                     Fondazione Macchi, ASST Sette Laghi, Varese, Italy;           FROM COAGULATION TO ANGIOGENESIS:
                     *Equally contributed                                          POTENTIAL ROLE OF FVIII IN ENDOTHELIAL
                                                                                   FUNCTIONALITY
                     Background: Venous thromboembolism (VTE) is char-
                     acterized by a multifactorial pathogenesis that often         A. Cucci1, C. Olgasi1, C. Borsotti1, G. Walker1,
                     occurs in the presence of provoking factors; major risk       I. Molineris2, F. Anselmi2, S. Assanelli1, C. Sgromo1,
                     factors induced provoked VTE (pVTE) whereas weakly            S. Oliviero2, A. Follenzi1
                     provoked (wpVTE) is associated with minor ones.
                                                                                   1Department of Health Sciences, Università del
                     Nevertheless, up to 50% of VTE events are defined as
                     “unprovoked” (uVTE) since their pathogenesis is large-        Piemonte Orientale, Novara, Italy; 2Italian Institute for
                     ly unknown, with a high risk of recurrence. In this           Genomic Medicine (IIGM), Università degli Studi di
                     regard, the presence of endothelial alterations deserves      Torino, Torino, Italy
                     to be explored as endothelial dysfunction (ED) is known
                     to trigger thrombus formation. Endothelial colony-form-       Background and Aims: Hemophilia A (HA) is a rare X-
                     ing cells (ECFCs) are the true endothelial progenitors        linked bleeding disorder caused by absence or reduced
                     proposed as cell therapy product or liquid biopsy in vas-     activity of coagulation FVIII. The extent of clinical man-




                                                                                                            ly
                     cular disorders. Aims: To investigate whether ECFCs in        ifestations of HA patients depends on the severity of the
                     VTE patients show alterations that may reflect a pertur-      FVIII deficiency. The main clinical manifestation is




                                                                                                    on
                     bation of the endothelial compartment involved in             spontaneous bleeding episodes that primarily involve
                     uVTE pathogenesis. Methods: ECFCs were isolated               hemarthroses and intracranial hemorrhages. The factors
                     and expanded from peripheral blood mononuclear cells          that initiate hemorrhage are not known, and the onset of




                                                                                              e
                     obtained from 53 VTE patients (16 uVTE, 21 wpVTE,             hemorrhage is often a random event, occurring either
                     16 pVTE) and 20 sex- and age-matched healthy donors
                     (HDs). ECFCs were analyzed for efficiency of isolation,
                     cell viability and growth. Endothelial function was
                                                                                       us
                                                                                   spontaneously or after minimal injury. Standard replace-
                                                                                   ment therapies are ineffective in preventing the bleeding
                                                                                   episodes. It is well established that FVIII is mainly
                                                                                   secreted by endothelial cells (ECs), but the impairment of
                                                                                   al
                     assessed at baseline by analyzing: a) platelet deposition
                     and fibrin formation under flow conditions, assessed in       vessel stability in HA patients and a correlation between
                                                                           ci

                     thrombogenesis assay by confocal microscopy; b)               FVIII and endothelial functionality has never been
                     thrombin formation assessed by thrombin generation            explained. Therefore, we aimed to explore the role of
                                                                     er


                     assay (TGA) on ECFCs - a modification of Hamker’s             FVIII in endothelial stability and identify significant dif-
                     method. Expression levels of the adhesion molecule            ferences in HA and healthy ECs. Methods: iPSCs-
                                                               m



                     CD106 (VCAM-1) and the pro-coagulant molecule                 derived ECs, differentiated from HA patients or healthy
                     Tissue Factor (TF) were also assessed, by flow cytome-        donors, were used as EC model. HA-ECs were trans-
                                                   om




                     try. Results: ECFCs were isolated from the three sub-         duced with a lentiviral vector (LV) carrying the B domain
                     groups of VTE patients with a similar efficiency as HDs       deleted form of FVIII under the control of the vascular
                     in term of frequency and number of colonies. In pVTE,         endothelial cadherin promoter (LV-VEC.FVIII). To eval-
                                                -c




                     a trend in the reduction of the frequency of subjects who     uate differences in healthy, HA ECs and LV-VEC.FVIII-
                     gave origin to ECFCs was observed. Time of appearance         transduced HA ECs, RNA-Seq analysis was carried out
                                        on




                     was longer in VTE patients with the difference being          and modulated genes were validated by qPCR. Finally, in
                     significant only in wpVTE patients. VTE ECFCs                 vitro and in vivo endothelial functional assays were per-
                     showed a higher rate of early senescence (passage≤2),         formed to investigate HA ECs impairment. Results:
                                   N




                     with wpVTE group being characterized by the highest           RNA-Seq analysis revealed different gene expression
                     frequency. By performing in vitro functional assays, we       profile among healthy, HA and LV-VEC.FVIII HA ECs,
                     observed: a) an increased platelet deposition and fibrin      particularly highlighting the down regulation of several
                     formation in VTE patients than in HDs as assessed by          genes in HA compared to healthy ECs, suggesting an
                     thrombogenesis assay; b) a reduction of the lag phase         impairment in HA ECs stability. LV-VEC.FVIII HA ECs
                     and an increase in thrombin formation in VTE patients         showed an intermediate transcriptional profile indicating
                     being more pronounced in uVTE patients as assessed by         only a partial correction after LV transduction.
                     TGA on ECFCs. CD106 and TF were barely expressed              Endothelial functional in vitro experiments demonstrated
                     on ECFCs in basal condition in all analyzed groups and        a reduced capability of HA ECs, showing a weakening in
                     only a slight increase in their expression was observed in    vessel-formation capability, migration potential and per-
                     ECFCs obtained by wpVTE patients. Conclusions: Our            meability when compared to healthy ECs. The impaired
                     results indicate alterations in the growth and in function-   phenotype was attenuated in LV-VEC.FVIII transduced
                     ality of VTE ECFCs thus confirming their use to inves-        HA ECs, suggesting the potential involvement of FVIII.
                     tigate ED role in uVTE pathogenesis. A deeper charac-         Finally, in vivo experiments in a HA mouse model
                     terization of ECFCs, still ongoing, may highlight the         demonstrated an altered permeability and tubulogenesis
                     molecular mechanisms involved in ED in uVTE and to            potential of HA vessels compared to wild type mice.
                     disclose whether they are associated with uVTE only are       Conclusions: These preliminary results, if confirmed in
                     shared with secondary VTE.                                    primary ECs, can provide new insights into the possible




         [page 20]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 21



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    extra-coagulative role of FVIII and it can be crucial to     pendent experiments (WT CD31+%: 70.3±11; HA
                    understand the key molecular targets missing in HA           CD31+%: 70.6±5, p value=0.9). Microglia contami-
                    patients at the cellular level impairing EC functionality.   nants (CD45+CD11b+CD31-) were eliminated by 5
                    This information can lead to new therapeutic approaches      days of culture in endothelial medium, with the persis-
                    resulting in a safer and more efficient treatment of HA.     tency of only 5% of pericytes (αSMA+ cells). No sig-
                                                                                 nificant morphological differences were appreciated
                                                                                 between WT and HA BMECs when cultured in FBS.
                    OC029                                                        However, qPCR analysis indicated that the absence of
                                                                                 FVIII secreted by BMECs modulates the expression of
                    THE ROLE OF MICROENVIRONMENT IN
                                                                                 endothelial markers. On the contrary, cells cultured
                                                                                 with WT or HA serum showed morphological and tran-
                    MORPHOLOGICAL AND TRANSCRIPTIONAL
                                                                                 scriptional changes with an increased rate of growth of
                    CHANGES OF BRAIN MICROVASCULAR                               WT BMECs cultured in WT murine serum compared to
                    ENDOTHELIAL CELLS IN WILD TYPE AND                           HA cells cultured in HA murine serum. Conclusions:
                    HEMOPHILIC MICE                                              These preliminary data indicate that WT and HA
                                                                                 BMECs have similar morphology and rate of growth if
                    S. Assanelli, C. Sgromo, P.E. Di Simone, C. Borsotti,        cultured in FBS but they showed a partially different
                    C. Olgasi, A. Follenzi                                       gene expression profile. This difference is marked when
                                                                                 cells are cultured in WT or HA serum suggesting that




                                                                                                          ly
                    Department of Health Sciences, Università del Piemonte       HA pro-inflammatory microenvironment could induce
                    Orientale, Novara, Italy




                                                                                                  on
                                                                                 a disorganized endothelium. These preliminary data
                                                                                 need to be further confirmed and could subtend a possi-
                    Background and Aims: Hemophilia A (HA) is a reces-           ble impaired vascular functionality in HA which could
                    sive X-linked bleeding disorder caused by the deficien-      justify the spontaneous ICHs in HA patients.




                                                                                            e
                    cy of the coagulation factor VIII (FVIII). Clinical man-
                    ifestations are bleeding episodes that primarily consist
                    in hemarthroses and intracranial hemorrhages (ICHs).
                    Standard therapies are ineffective in preventing the
                                                                                     us
                                                                                 OC030
                    bleeding episodes that can occur without any clear
                                                                                 al
                    cause. The manifestation of ICHs is often spontaneous,       P2Y12 DEPENDENT MODULATION OF
                                                                         ci

                    and mainly occurs in patients that do not adhere to the      PLATELET-ASSOCIATED TISSUE FACTOR AND
                    prophylaxis regimen with a mortality around 20%.             INTRACELLULAR LOCALIZATION
                                                                   er


                    Although the cause of the development of ICHs is still
                    unknown, a correlation between the severe form of HA         M. Brambilla1, P. Canzano1, G.E. Rovati2, M. Conti1,
                                                             m



                    and ICHs in children has been described, emphasizing         A. Becchetti1, A. Loffreda3, M. Cattaneo4,
                    the importance of prophylaxis in reducing the risk of        E. De Candia5, G.M. Podda4, C. Pinna2, E. Tremoli6,
                                                 om




                    ICHs. Additionally, clinical evidence indicate a general     M. Camera1,2
                    microvascular endothelial impairment in HA patients
                                                                                 1Centro   Cardiologico Monzino IRCCS, Milan, Italy;
                    and that HA untreated patients show a pro-inflammato-
                                                                                 2Dept   of Pharmaceutical Sciences, Università degli
                                              -c




                    ry environment with higher levels of cytokines in the
                    plasma compared to healthy controls. Thus, we aim to         Studi di Milano, Milan, Italy; 3Experimental Imaging
                                       on




                    investigate morphological and transcriptional differ-        Center, San Raffaele Scientific Institute, Milan, Italy;
                                                                                 4Unità di Medicina II, ASST Santi Paolo e Carlo, Dept
                    ences in brain microvascular endothelial cells (BMECs)
                    isolated from wild type (WT) and HA mice and to              of Scienze della Salute, Università degli Studi di Milano,
                                  N




                    define if WT or HA microenvironment can influence            Milan, Italy; 5Dept of Traslational Medicine and
                    the phenotype of BMECs. Methods: To isolate murine           Surgery, Università Cattolica del Sacro Cuore, Rome,
                    BMECs from WT and HA C57BL/6 mice brains were                Italy; 6Maria Cecilia Hospital, Cotignola, Italy
                    removed from 8 weeks old mice, and they were
                    mechanically disaggregated. After enzymatic digestion        Background and Aims: ADP is stored at high concen-
                    of the tissues, myelin was eliminated, and endothelial       trations in platelet dense granules and it enables platelet
                    cells were separated from the debris through a Percoll®      activation and aggregation by binding to two purinergic
                    gradient. FACS analysis was performed to evaluate            receptors on the platelet surface: P2Y1, triggering platelet
                    endothelial markers expression. BMECs were then cul-         activation, and P2Y12, which amplifies the process poten-
                    tured in endothelial medium with FBS, WT and HA              tiating platelet secretion through membrane exposure of
                    murine serum. After 5 days of culture, qPCR and              several intracellular proteins, including Tissue Factor
                    immunofluorescences were performed to verify the             (TF). The contribution of P2Y1 and P2Y12 receptors in
                    expression of endothelial markers and to reveal the          platelet TF modulation is still unknown. The aims of the
                    presence of other cell types. Results: Isolation of          study were to assess: 1) the involvement of P2Y1 and
                    BMECs was optimized for both WT and HA mice.                 P2Y12 receptors in ADP-induced TF exposure on the
                    Most of the live isolated cells were endothelial             platelet surface in healthy subjects’ platelets, exploiting
                    (CD31+CD45-): the number of isolated CD31+ cells             platelets from a P2Y12-deficient patient; 2) TF intracellu-
                    was not different between WT and HA mice in 3 inde-          lar localization through immunogold-electron




                                                              [Hematology Reports 2021; 13:s3]                                                  [page 21]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 22



         Oral communications




                     microscopy and pharmacological approach, taking                A. De Curtis1, T. Panzera1, M. Ninivaggi2, C. Cerletti1,
                     advantage from the study of two Grey Platelet Syndrome         G. de Gaetano1, L. Iacoviello1,4, M.B. Donati1,
                     (GPS) patients’ platelets. Methods: TF+- and P-selectin+-      B. de Laat2,5, for the Moli-sani Study investigators
                     platelets were evaluated by whole blood (WB) flow
                                                                                    1Department  of Epidemiology and Prevention, IRCCS
                     cytometry. In vitro experiments were performed on 10
                     healthy subjects (HS; n=5 males and n=5 females; mean          Neuromed, Pozzilli, IS, Italy; 2Department of Data
                     age 39±6y) and WB was incubated with AR-C69931MX               Analysis and Artificial Intelligence, Synapse Research
                     (1pM–100nM) or MRS-2500 (1pM–100nM) and stimu-                 Institute, Maastricht, the Netherlands; 3Mediterranea
                     lated with ADP (10µM). To assess actin and microtubular        Cardiocentro, Napoli; 4Research Center in
                     polymerization involvement in TF and P-selectin expres-        Epidemiology and Preventive Medicine (EPIMED),
                     sion, platelets were preincubated with cytochalasin D          Department of Medicine and Surgery, University of
                     (10µM) or colchicine (10µM). Immunogold labelling              Insubria, Varese, Italy; 5Department of Functional
                     and electron microscopy were also performed to define          Coagulation, Synapse Research Institute, Maastricht, the
                     platelet TF localization. Two patients with GPS syn-           Netherlands
                     drome (males, aged 31y and 15y) and a subject with
                     inherited severe P2Y12 deficiency (male, aged 65y) were        Background and Aims: Thrombin is the key enzyme
                     also studied to confirm our hypothesis. Results: Flow          in the coagulation process. It leads to clot formation by
                     cytometry analysis of WB from HS showed that P2Y12-            promoting activation of platelet and coagulation fac-
                     inhibitor AR-C69931MX, but not P2Y1-inhibitor MRS-             tors. On the other hand, thrombin bound to thrombo-




                                                                                                           ly
                     2500, concentration-dependently prevented ADP-stimu-           modulin activates protein C, an inhibitor of the coagu-
                                                                                    lation cascade. The Thrombin generation (TG) assay




                                                                                                    on
                     lated TF exposure. To confirm the exclusive involvement
                     of P2Y12 in TF expression, we analyzed blood of a              assesses the potential of plasma to form thrombin, and
                     P2Y12-deficient patient. Upon ADP stimulation, no              it is useful for detecting hyper-or-hypocoagulability
                     increase of TF expression on the platelet surface was          tendency at the individual level. To date a limited num-




                                                                                              e
                     observed, but TF was readily detectable after stimulation      ber of studies have investigated the relationship of TG
                     with U46619 (+1.7 fold) or TRAP-6 (+1.4 fold). Unlike
                     what observed for TF, the membrane exposure of P-
                     selectin, a protein stored in platelet alpha-granules, was
                                                                                       us
                                                                                    with the risk of coronary heart disease (CHD) and
                                                                                    stroke, with inconsistent results. The possible associa-
                                                                                    tion of both low and high values of the TG parameters
                                                                                    al
                     significantly modulated by both P2Y1 and P2Y12. This           with the incidence of CHD and stroke was therefore
                     finding led us to hypothesize a different cellular localiza-   evaluated in a large cohort of an Italian adult popula-
                                                                            ci

                     tion of these proteins. Inhibition of open canalicular sys-    tion. Methods: We analysed data on 20,109 individu-
                     tem (OCS) externalization, by preincubation with               als (53.3% women, mean age 54.8 ±11.5 years)
                                                                      er


                     cytochalasin D or colchicine, impaired ADP-induced TF          enrolled in the Moli-sani study (2005-2010), without a
                     but not P-selectin expression, supporting TF storage           history of cardiovascular disease or use of vitamin K
                                                                m



                     within OCS as indicated also by immunogold labelling           antagonists at baseline, for whom TG data were avail-
                     and electron microscopy analysis. Furthermore, we con-         able. TG curves were measured in each sample at 5 pM
                                                   om




                     firmed this hypothesis in patients with GPS syndrome, a        tissue factor by the calibrated automated thrombinog-
                     disease characterized by large platelets lacking normal        raphy method (Synapse Research Institute, The
                     alpha-granules. Flow cytometry analysis showed signifi-        Netherlands). The whole cohort was followed-up to
                                                -c




                     cantly reduced levels of P-selectin+-platelets (about -        December 2015 for fatal or non-fatal CHD and stroke
                     65%) compared to HS. Conversely, platelet-associated           events; the latter were ascertained by record linkage to
                                        on




                     TF expression in GPS patients was comparable to that of        the regional hospital discharge form and death reg-
                     HS, despite the alpha-granule defect. Conclusions: This        istries and validated by checking medical records or
                     study shows for the first time 1) the unique involvement       death certificate, respectively. Multivariable hazard
                                   N




                     of P2Y12 receptor in ADP-induced TF exposure and 2)            ratio (HR) and 95% confidence interval (CI) for out-
                     the TF intracellular localization that is stored in the OCS    comes were estimated by Cox-proportional hazard
                     and exposed upon stimuli-induced OCS externalization.          models, categorizing TG parameters in quintiles (the
                     All together our findings add new insights into the regu-      third acted as the reference) including a large panel of
                     lation of the procoagulant activity of platelets highlight-    cardiovascular risk factors as possible confounders.
                     ing that as for phosphatylserine, also TF exposure is a        Results: Over a median follow-up of 8.2 years, a total
                     P2Y12 dependent mechanism.                                     of 571 (2.8%) CHD and 183 (0.9%) stroke events were
                                                                                    ascertained. No association between any TG parameter
                                                                                    and CHD was found. In contrast, individuals in the
                     OC031                                                          lowest or the highest endogenous thrombin potential
                                                                                    (ETP) quintiles showed an increased risk of total stroke
                                                                                    compared to the third one: multivariable HR=1.63
                     THROMBIN GENERATION IS ASSOCIATED WITH                         (95%CI: 1.03-2.58) and HR=1.75 (95%CI: 1.06-2.89),
                     STROKE BUT NOT WITH CORONARY HEART                             respectively (Table 1). Dose-response analyses by use
                     DISEASE IN A GENERAL ADULT POPULATION:                         of restricted cubic spline functions, confirmed the U-
                     THE MOLI-SANI STUDY                                            shaped relationship between ETP and risk of total
                                                                                    stroke (P value for overall association 0.032; P value
                     S. Costanzo1, R. de Laat-Kremers2, A. Di Castelnuovo3,         for non-linear association 0.013). Preliminary, under-




         [page 22]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 23



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    powered analyses by stroke subtypes showed U-shaped         stable anticoagulation in the range from 1.5 to 4.5 INR.
                    association of ETP with ischemic (N=95), but a nega-        These patients are difficult to be recruited as the con-
                    tive linear link of ETP with haemorrhagic (N=50)            ventional therapeutic intervals for VKA is from to 2.0 to
                    events. Conclusions: In comparison with median val-         3.0 INR. A possible solution could be including in the
                    ues, both lower and higher levels of ETP, correspon-        calibration patients with less intense anticoagulation.
                    ding to the total amount of thrombin formed during the      We sought to investigate the impact of the above sim-
                    assay, are associated with an increased risk of total       plified procedure on the following parameters of cali-
                    stroke. The thrombin dualistic role in the coagulation      bration: (i) ISI; (ii) coefficient of variation (CV) of the
                    cascade could be an explanation of the U-shaped rela-       calibration and (iii) INR. Methods: We used eight data
                    tionship observed with total (particularly ischemic)        sets of previous calibrations of a rabbit thromboplastin
                    stroke. Further confirmation of a relationship between      (Sclavo PT Ready Quick, Dasit, Milano, Italy) in com-
                    TG and CHD/stroke would not only contribute to the          bination with the coagulometer CSA 5100, Sysmex,
                    knowledge on the physio-pathological processes lead-        Kobe, Japan. The above calibrations were performed
                    ing to thrombosis, but also suggest new preventive          against the rabbit thromboplastin WHO standard (coded
                    approaches.                                                 RBT/16) and included patients on anticoagulation as
                                                                                required by WHO and with INR from 1.5 to 4.5.
                    Table 1.                                                    Parameters of calibration as determined with the full
                                                                                data sets were identified as “full calibrations” and were
                                                                                considered as reference. Each of the data sets were then




                                                                                                        ly
                                                                                used to recalculate the calibration parameters after
                                                                                including patients with INR <4.0, <3.5 or <3.0, which




                                                                                                 on
                                                                                were identified as “trimmed calibrations” and compared
                                                                                to those from the “full calibrations”. Results: Results
                                                                                are in Table 1.




                                                                                           e
                                                                                    us
                                                                                Table 1.
                                                                                al
                                                                        ci
                                                                  er
                                                             m
                                                 om




                    OC032
                                              -c




                    THROMBOPLASTIN CALIBRATION REVISITED TO
                    LOOK FOR POSSIBLE REVISION OF THE WORLD
                                      on




                    HEALTH ORGANIZATION RECOMMENDATIONS

                    E. Scalambrino1, M. Clerici1, V. Chantarangkul1,
                                  N




                    F. Peyvandi1,2, A. Tripodi1
                    1Fondazione   IRCCS Ca’ Granda Ospedale Maggiore
                    Policlinico, Angelo Bianchi Bonomi Hemophilia and
                    Thrombosis Center, Milano and Fondazione Luigi Villa,
                    Milano, Italy; 2Università degli Studi di Milano,
                    Department of Pathophysiology and Transplantation,
                    Milano, Italy

                    Background and Aims: Monitoring vitamin K antago-
                    nists (VKA) entails frequent measurement of the pro-
                    thrombin time (PT) with results expressed as interna-
                    tional normalized ratio (INR) to be used for dose adjust-   There was marginal variation of the ISI, CV and INR
                    ment. To this end thromboplastin calibration is essential   that can hardly be considered of practical significance.
                    to determine the international sensitivity index (ISI)      CV was the most affected parameter, which increased
                    required to calculate the INR. The procedure for cali-      from the full to the trimmed <3.0 calibration, but never
                    bration recommended by the World Health                     exceeded the 3% cut off value recommended by WHO.
                    Organization (WHO) calls for selection of patients on       Conclusions. Should the results of this pilot study be




                                                             [Hematology Reports 2021; 13:s3]                                                 [page 23]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 24



         Oral communications




                     confirmed      for      calibrations   of     other           (Chromogenix Coamatic® Factor VIII and Trinichrom
                     thromboplastin/coagulometer combinations, the revi-           FVIII Stago). Each sample was also tested using throm-
                     sion of the WHO recommendations to include patients           bin generation assay (TGA) by ST Genesia system –
                     with INR from 1.5 to 4.0 is warranted.                        Stago which measured the generated thrombin in fluo-
                                                                                   rescence. The parameters considered were normalized:
                                                                                   lag-time, peak height, endogenous potential of active
                     OC033                                                         thrombin (ETP), velocity index, time to peak and start
                                                                                   tail. Results: The classical OSA assay could show sig-
                     EXTENDEND-HALF-LIFE RECOMBINANT FVIII
                                                                                   nificant discrepancies between the certificated and real
                                                                                   EHL-FVIII concentrations. The determination of
                     PRODUCTS (EHL-FVIIIS): ONE STAGE
                                                                                   Efmoroctocog alfa by OSA provides a value two-fold
                     COAGULATION ASSAY OR CHROMOGENIC                              higher than the expected level. The calibrated-OSA
                     ASSAY?                                                        assay is more accurate in determining the values of all
                                                                                   EHL-FVIIIs, showing in the spiked experiments a drug
                     M. Sacco1, S. Lancellotti2, M. Tardugno1,                     recovery from 92% to 101%. On the contrary, the CSA,
                     R. De Cristofaro1,2                                           despite has been used to determine the potency of all
                     1Catholic
                                                                                   products, reported a CV more than 30%. Noteworthy,
                                University, Translational Medicine and             Damoctocog alpha-pegol levels were significantly
                     Surgery Department Rome; 2Fondazione Policlinico              underestimated by one of the CSA tested, underestimat-




                                                                                                           ly
                     Universitario AGemelli, IRCCS, Center for                     ing the nominal value up to 50%. TGA showed a signif-
                     Hemorrhagic and Thrombotic Diseases and




                                                                                                   on
                                                                                   icant functional difference between EHL-FVIII prod-
                     Haemophilia Center, Rome, Italy                               ucts. The thrombogram obtained from spiked EHL-
                                                                                   FVIIIs, demonstrated lower sensitivity at concentration
                     Background and Aims: The extended-half-life recom-            <0.1 IU/dL. Conclusions: In this scenario, a standard




                                                                                              e
                     binant FVIII products (EHL-FVIIIs) have been intro-           calibration curve for each EHL-FVIII product could
                     duced to improve the FVIII pharmacokinetic profile
                     (increasing about 1.5 fold the half-life of FVIII) reduc-
                     ing the infusions number in hemophilic patients and the
                                                                                       us
                                                                                   avoid inaccurate laboratory results, which might heavi-
                                                                                   ly impact on patient management.
                     annual bleeding rates with less frequent dosing and
                                                                                   al
                     higher trough plasma concentrations. These products           OC034
                                                                           ci

                     are structurally and functionally heterogeneous; in fact,
                     chemical coupling of Polyethylene glycol (PEG)
                                                                     er


                     increases hydrodynamic volume to prevent FVIII clear-         COAGUCHEK® PRO II PERFORMANCE
                     ance, while the fusion with other proteins, such as the       EVALUATION TO ASSESS DIRECT ORAL
                                                               m



                     Fc fragment of IgG1 or albumin, protects FVIII from           ANTICOAGULANT (DOAC) ACTION.
                     lysosomal sorting and degradation. These different            THE DOAC-CHECK STUDY
                                                   om




                     strategies lead to several laboratory discrepancies in
                     FVIII coagulant (FVIII:C) measurement by chro-                C. Legnani1, M. Cini1, S. Testa2, A. Tosetto3,
                     mogenic substrate assay (CSA) and one-stage clotting          C. Dellanoce2, S. Bellesso3, G. Carli3, I. Nichele3,
                                                                                   L. Lissandrini3, S. Zorzi1, E. Antonucci1, G. Palareti1
                                                -c




                     assay (OSA) inducing an altered FVIII recovery with an
                     over/under-estimating by more than 30%. Similar dis-
                                                                                   1Arianna
                                        on




                     crepancies were observed in patients receiving                            Anticoagulazione Foundation, Bologna;
                     ReFacto® (B-Domain-Deleted FVIII) a decade ago and            2Haemostasis    and Thrombosis Center, Laboratory
                     has been resolved with the use of specific ReFacto            Medicine Department, Cremona Hospital, Cremona;
                                   N




                     Laboratory Standard. The laboratory assays used               3Department of Hematology, San Bortolo Hospital,

                     should obtain the recovery of FVIII:C closely in accor-       Vicenza, Italy
                     dance with the assay used to demonstrate the product
                     efficacy during clinical trials. The aim of our study is to   Background and Aims: Direct oral anticoagulants
                     evaluate, with four different types of assays, FVIII:C        (DOACs) do not require routine laboratory monitoring,
                     and recovery of spiked EHL FVIIIs in FVIII-deficient          which is suggested in specific conditions, especially
                     human plasma. Methods: The EHL-FVIII products                 emergencies. Specific tests for DOAC are available,
                     tested were: Damoctocog alpha-pegol, Rurioctocog              though implemented in few laboratories. PT and PTT
                     alpha-pegol,       Turoctocog        alpha-pegol       and    may be useful for a rough evaluation of anticoagulation
                     Efmoroctocog alfa. All EHL-FVIIIs were reconstituted          level but are performed in laboratory, and results are
                     according to the manufacturer’s instructions and serial       delayed by sample transportation and handling.
                     dilutions (1/0.5/0.1/0.05/0.025 UI/dL) were obtained by       Conversely, POCT for PT and PTT measurements are
                     using HemosIL® FVIII Deficient Plasma (Werfen). For           currently used. The DOAC-CHECK Study, an observa-
                     each product, FVIII:C was then tested through OSA             tional, national, multicentre, no-profit study, aimed to
                     (Werfen) by using two different APTT reagents contain-        evaluate weather CoaguChek® Pro II (Roche
                     ing different plasma activators (colloidal silica and         Diagnostics) use can provide reliable information in
                     Ellagic Acid), EHL-FVIII-calibrated OSA methods and           DOAC treated patients. The study, promoted by
                     two different commercially available CSA                      Fondazione Arianna Anticoagulazione, was supported by




         [page 24]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 25



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Roche Diagnostics and Fondazione Cassa di Risparmio           Hemorrhagic and Thrombotic Diseases and
                    di Bologna. Methods: The study was carried out in two         Haemophilia Center, Rome; 3Fondazione Policlinico
                    FCSA (Italian Federation of Thrombosis Centers) cen-          Universitario AGemelli, IRCCS, Department of
                    ters. PT and PTT were performed by CoaguChek® Pro II          Digestive, Endocrine and Metabolic System, Institute of
                    using capillary blood; at the same time venous samples        Internal Medicine and Geriatrics, Rome, Italy
                    were collected for measuring DOAC levels by chro-
                    mogenic assay (Diagnostica Stago). Since data to define       Background: Portal vein thrombosis (PVT) represents
                    relevant DOAC concentrations are currently limited we         one of the most fearsome comorbidity of cirrhosis and
                    choose 3 different concentration thresholds for our analy-    its prevalence is higher in patients with decompensated
                    sis (30, 50 and 100 ng/mL). Using the ROC curves, for         disease. These patients are characterized by a fragile
                    each test/DOAC threshold, as the ideal cut-off point was      hemostatic balance with a decrease of both procoagu-
                    selected the one that yielded a sensitivity of at least 95%   lant and anticoagulant factors, synthesized by hepato-
                    (misprediction percentage <5%, considered sufﬁciently         cytes, together with an increase in endothelial-derived
                    safe for clinical application) associated to the highest      factors such as Factor VIII (FVIII) and von Willebrand
                    possible speciﬁcity, to avoid false-negative results but      factor (VWF). Simultaneously, the pro-fibrogenic trans-
                    simultaneously to identify the largest number of patients     formation of liver’s stellate cells causes a progressive
                    eligible for emergency treatment such as thrombolysis or      decrease of ADAMTS-13 levels, while VWF multimers
                    emergency surgery. Results: 512 patients were enrolled;       secretion by endothelial cells is strongly enhanced. This
                    222 received Apixaban, 120 Edoxaban, 111                      unbalance induces an accumulation of ultra-large VWF




                                                                                                         ly
                    Rivaroxaban, and 59 Dabigatran. For Edoxaban and              multimers in sinusoidal circulation where PVT in intra-
                    Rivaroxaban both for CoaguChek® Pro II PT and PTT a           and extra-hepatic branches could occur, especially in




                                                                                                  on
                    sensitivity >95% corresponded to satisfying specificity       decompensated cirrhosis. A decreased portal vein flow
                    values, especially at the 50 and 100 ng/mL threshold          velocity <15cm/sec2, thrombophilia factors and portal
                    DOAC concentrations; the NPVs resulted in the range           hypertension are well-known risk factors predisposing




                                                                                            e
                    90-100% both for CoaguChek® Pro II PT and PTT at all          to PVT but it is still unclear how the primary and sec-
                    DOAC thresholds. Conversely, CoaguChek® Pro II PT
                    and PTT could identify patients with Apixaban concen-
                    trations above the pre-defined thresholds, but only at the
                                                                                     us
                                                                                  ondary haemostatic abnormalities are involved in PVT
                                                                                  development. Thus, the identification of biomarkers
                                                                                  acting as predictors of PVT in compensated cirrhotic
                                                                                  al
                    expense of a high number of false-positive cases, thus        patients could have a significant clinical impact allow-
                    excluding from thrombolysis/surgery a too large number        ing to identify patients at risk who could eventually
                                                                          ci

                    of patients. CoaguChek® Pro II PT and PTT do not seem         undergo prophylactic therapy. This observational
                    to be useful for identifying Dabigatran concentrations        prospective clinical trial on compensated cirrhotic
                                                                    er


                    higher than the pre-defined threshold; acceptable results     patients (ClinicalTrial.gov: NCT03322696) could iden-
                    were obtained only for CoaguChek® Pro II PTT at the           tify ADAMTS-13/VWF:GpIbR ratio as a key predic-
                                                              m



                    threshold of 100 ng/mL Dabigatran concentration.              tive factor of PVT development in cirrhotic patients.
                    Conclusions: Our results suggest that CoaguChek® Pro          Methods: The final 79/118 enrolled subjects were fol-
                                                  om




                    II can be used to qualitatively identify relevant concen-     lowed for 48 months and underwent Doppler-ultra-
                    trations of Edoxaban and Rivaroxaban, but not of              sound liver examination and clinical (Child-Pugh and
                    Apixaban and Dabigatran. However, as the suggested            MELD-score) and hematochemical (PT%, APTT%,
                                               -c




                    cut-offs were determined retrospectively, further evalua-     AT%, PC%, PS%, LAC, fibrinogen, FVIII:C, D-dimer)
                    tion in a prospective clinical trial, ideally in emergency    assessment. VWF antigen and VWF:GpIbR were meas-
                                       on




                    situations, is warranted to investigate the clinical safety   ured by chemiluminescence assays; ADAMTS-
                    of this approach.                                             13:activity by a FRET-based-assay; both FV Leiden
                                                                                  G1691A and the prothrombin G20210A polymor-
                                  N




                                                                                  phisms by using the RT-PCR; statistical analyses by the
                    OC035                                                         SPSS-software. Results: Five patients presented PVT,
                                                                                  the cumulative incidence of PVT in this cohort was:
                                                                                  2.5, 3.8, and 6.3% at 1, 2 and 3 years after the enroll-
                    ADAMTS-13/VON WILLEBRAND RATIO:                               ment. These patients showed a statistically significant
                    A PROGNOSTIC BIOMARKER FOR PORTAL VEIN                        reduction of ADAMTS-13/VWF:GpIbR ratio com-
                    THROMBOSIS (PVT) IN COMPENSATED LIVER                         pared to patients without PVT [mean value: 0.30 (0.28-
                    CIRRHOSIS. A PROSPECTIVE OBSERVATIONAL                        0.32) vs 0.69 (0.61-0.77). Univariate analysis showed
                    STUDY                                                         that at the enrollment visit ADAMTS-13 levels,
                                                                                  ADAMTS-13/VWF:GpIbR ratio, PS, PC, Platelets
                    M. Sacco1, M. Tardugno1, S. Lancellotti2, A. Ferretti2,       count, INR were significantly correlated with PVT. The
                    F.R. Ponziani3, L. Riccardi3, M.A. Zocco3,                    ADAMTS-13/VWF:GpIbR ratio sensitivity and speci-
                    A. De Magistris3, F. Santopaolo3, M. Pompili3,                ficity as PVT predictor, 86% and 80% respectively,
                    R. De Cristofaro1,2                                           were assessed by a ROC curve (Figure 1), highlighting
                                                                                  the AUC equal to 0.833 and a cut-off-value=0.40,
                    1Catholic  University, Translational Medicine and             below which the risk to develop a PVT complication
                    Surgery Department Rome; 2Fondazione Policlinico              increases (OR 14.6, 95% C.I.:1.36-157.2, p=0.027) as
                    Universitario AGemelli, IRCCS, Center for                     also confirmed by a Cox-regression-analysis, where the




                                                               [Hematology Reports 2021; 13:s3]                                               [page 25]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 26



         Oral communications




                     risk to develop a PVT rises exponentially after 30th        and Internal Medicine, Policlinico San Marco, Zingonia,
                     month-follow-up in patients with a ADAMTS-                  Bergamo; 8Oncology Unit, Hospital San Giovanni
                     13/VWF:GpIbR ratio <0.4 compared to patients with           Addolorata, Rome; 9DIPO, ASST Papa Giovanni XXIII,
                     ratio >0.4 [HR:7.7 (C.I.; 2.2-26.7, p=0.001].               Bergamo; 10School of Medicine, University of Milan-
                     Conclusions: In compensated cirrhosis a ADAMTS-             Bicocca, Milan, Italy
                     13/VWF:GpIbR ratio <0.4 could be a useful predictive
                     biomarker for the identification and stratification of      Background and Aims: Gastrointestinal (GI) cancers
                     patients at risk of PVT development. Further studies are    represent 26% of the global cancer incidence and more
                     needed to confirm these findings even in moderate and       than one-third of all cancer-related deaths. The clinical
                     severe clinical setting where new treatment strategy        benefits of treatments developed over years differ criti-
                     could be used for preventing and treating this complica-    cally among individuals and, in many cases, patient’s
                     tion of cirrhosis.                                          prognosis remains poor. Due to the reciprocal interac-
                                                                                 tion between coagulation and cancer, the biomarkers of
                                                                                 the hemostatic system could be useful in monitoring
                                                                                 prognosis and in the choice of the most appropriate
                                                                                 treatment. In a prospective cohort of patients with
                                                                                 newly diagnosed metastatic GI cancer, we aimed to
                                                                                 evaluate whether pre-chemotherapy hemostatic bio-
                                                                                 markers may predict for 6 months disease progression




                                                                                                         ly
                                                                                 (6m-DP), and for 1-year overall survival (1-year OS).
                                                                                 Methods: Prospectively enrolled patients with metasta-




                                                                                                 on
                                                                                 tic GI cancer from the HYPERCAN study were ana-
                                                                                 lyzed. At enrollment, before starting any curative
                                                                                 chemotherapy, plasma samples were collected and test-




                                                                                            e
                                                                                 ed for thrombin generation (TG) potential and D-dimer,

                                                                                     us
                                                                                 fibrinogen, and prothrombin fragment 1+2 (F1+2) lev-
                                                                                 els. DP and mortality were monitored during follow-up.
                                                                                 Results: A cohort of 626 GI cancer patients (age: 66 y
                                                                                 al
                                                                                 [26-87]), 462 with colorectal and 164 with gastric can-
                                                                                 cers, was available for analysis. After 6 months from
                                                                         ci

                                                                                 the start of chemotherapy, DP occurred in 148 patients,
                                                                                 providing a cumulative incidence of 24.8% (CI 95%
                                                                   er


                                                                                 21.4-28.4) with death as competing risk. Baseline D-
                                                                                 dimer levels were significantly (p=0.001) higher in
                                                              m



                                                                                 these patients compared to those without DP. Gastric
                                                                                 cancer, pre-chemotherapy D-dimer, ETP, and the pres-
                                                  om




                     Figure 1.                                                   ence of more than 1 metastatic sites were identified as
                                                                                 independent risk factors for 6m-DP by multivariate
                                                                                 Fine-Gray proportional hazard regression analysis.
                                               -c




                     OC036                                                       After 1-year, overall survival (OS) was 75.7% (CI 95%
                                                                                 71.9-79.0). Patients who died showed higher baseline
                                       on




                                                                                 D-dimer (p<0.001), F1+2 (p=0.004), fibrinogen
                     BIOMARKERS OF HYPERCOAGULABILITY AND                        (p=0.023), and TG ETP (p<0.01) values compared to
                     PREDICTION FOR EARLY DISEASE                                surviving. Multivariate Cox regression identified inde-
                                   N




                     PROGRESSION AND MORTALITY IN PATIENTS                       pendent risk factors for 1-year OS the followings: gas-
                     WITH METASTATIC GASTROINTESTINAL CANCER                     tric cancer, D-Dimer, ETP, ECOG-PS, and advanced
                                                                                 cancer stage at diagnosis. D-dimer and TG ETP levels
                     C. Giaccherini1, M. Marchetti1, C. Verzeroli1, L. Russo1,   were dichotomized using ROC curves and a score was
                     S. Gamba1, C.J. Tartari1, S. Bolognini1, C. Ticozzi1,       created accordingly for classification of patients at low,
                     F. Schieppati1, A. Santoro2, F. De Braud3, G. Gasparini4,   intermediate, or high risk of early DP. As a result, hav-
                     F. Petrelli5, F. Giuliani6, A. D’Alessio7, M. Minelli8,     ing at least one or both parameters above the specific
                     R. Labianca9, A. Falanga1,10 on behalf of the HYPERCAN      threshold was a risk factor not only for 6m-DP (inter-
                     Investigators*                                              mediate vs low: SHR=2.1 [1.3-3.4]; high vs low:
                                                                                 SHR=2.3 [1.4-3.9]), but also for 1-year mortality (inter-
                     1Immunohematology      and Transfusion Medicine,            mediate vs low: HR=2.7 [1.5-4.8]; high vs low: HR=5.5
                     Hospital Papa Giovanni XXIII, Bergamo; 2Oncology            [3.1-9.8]). Conclusions: Our results showed that, in
                     Unit, IRCCS National Cancer Institute, Milan;               newly diagnosed metastatic GI cancer patients, pre-
                     3Oncology Unit, IRCCS Humanitas Institute, Rozzano,         treatment TG potential and D-dimer appear as promis-
                     Milan; 4Oncology Unit, Hospital San Filippo Neri,           ing candidate biomarkers to predict for 6-month DP and
                     Rome; 5Oncology Unit, Hospital Treviglio-Caravaggio,        1-year OS. In this setting, the role of TG as a prognostic
                     Treviglio, Bergamo; 6Oncology Unit, IRCCS Cancer            biomarker emerges for the first time in a large prospec-
                     Institute Giovanni Paolo II, Bari; 7Medical Oncology        tive cohort of GI cancer patients.




         [page 26]                                         [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 27



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    OC037                                                          L. Chatenoud3, G. Portella1, D. Poli4, S. Testa4,
                                                                                   R. Baiocchi1
                    ASSOCIATION BETWEEN NON-O BOOD TYPE AND                        1Emergency NGO ONLUS, Milan, Italy; 2SALAM Centre
                    EARLY UNEXPLAINED RECURRENT                                    for Cardiac Surgery, Khartoum, Sudan; 3Department of
                    SPONTANEOUS ABORTION IN WOMEN WITH AND                         Public Health, Laboratory of Clinical Epidemiology,
                    WITHOUT THROMBOPHILIA                                          Istituto di Ricerche Farmacologiche Mario Negri
                                                                                   IRCCS, Milan, Italy; 4Federazione Centri per la
                    L. Spiezia, A. Poretto, E. Borella, G. Turatti,                Diagnosi Della Trombosi e la Sorveglianza delle Terapie
                    M. Marobin, C. Simion, E. Campello, D. Tormene,                Antitrombotiche (FCSA), Milan, Italy; 5Università degli
                    P. Simioni                                                     Studi, Milan, Italy

                    Thrombotic and Hemorrhagic Diseases Unit, Padova               Background and Aim: Pregnancy on anticoagulants
                    University Hospital, Padova, Italy                             after mechanical heart valve replacement presents a high
                                                                                   risk of complications for both mother and baby.
                    Background and Aims: The association between inherit-          Moreover, pregnancy is a prothrombotic condition itself.
                    ed thrombophilias (IT) and recurrent spontaneous abor-         A safe and effective anticoagulation regimen for both
                    tion (RSA) has been studied extensively without reaching       mother and foetus doesn’t exist. The most effective
                    definitive conclusions. We retrospectively evaluated           drugs to prevent valve thrombosis are Vitamin K




                                                                                                           ly
                    whether non-O blood type – the commonest prothrombot-          Antagonists (VKAs): the dosage needs to be adjusted
                    ic risk factor – might be associated with a higher risk to     with frequent INR checks. VKAs can have embryopath-




                                                                                                   on
                    develop early unexplained RSA in women with and with-          ic and teratogenic effect; moreover they can cause foetal
                    out IT. Methods: N. 238 consecutive women with a his-          bleeding. Patients in follow-up and anticoagulant treat-
                    tory of early (i.e. before 12 weeks of gestation) objective-   ment at the Salam Centre for Cardiac Surgery in Sudan




                                                                                              e
                    ly unexplained RSA (i.e. ≥3 consecutively) tested for IT       live scattered in a large area, with limited access to the
                    at the laboratory of Thrombotic and Hemorrhagic
                    Diseases Unit, Padova University Hospital, Padova, Italy
                    between December 2011 and December 2021 were con-
                                                                                       us
                                                                                   Center. For all these reasons pregnancy is discouraged
                                                                                   and contraception and therapeutic abortion recommend-
                                                                                   ed, but this guidance frequently goes unheeded. The aim
                                                                                   al
                    sidered for enrollment. A group of healthy women, age-         is to analyze pregnancy outcomes in women with
                    matched, with ≥1 normal pregnancy and no history of            mechanical valve prosthesis followed by the
                                                                           ci

                    adverse pregnancy outcomes acted as controls. All partic-      Anticoagulant Clinic (AC) of Salam Centre. Methods:
                    ipants were tested for antithrombin, protein C and S defi-     Data were collected from all pregnancies recorded
                                                                     er


                    ciency, prothrombin G20210A, FV Leiden and MTHFR               among patients followed by our AC from April 2017
                    mutations and were ABO genotyping. Results: N. 216             until November 2021. Death, valve thrombosis and
                                                               m



                    (mean age at the time of first abortion 36 yrs, range 24-43    major bleeding occurred during pregnancy till 6 weeks
                    yrs) women were enrolled. No significant differences           after its conclusion have been considered as maternal
                                                  om




                    between cases and controls according to age and BMI (p         outcomes; miscarriage, intrauterine foetal death and
                    >0.05 in each comparison) were observed. Non-O blood           neonatal death have been considered as foetal outcomes.
                    type was significantly more prevalent in cases than con-       Results: 307 pregnancies in 253 women were assessed.
                                               -c




                    trols (p <0.05). Adjusted Odds ratio (aOR) of RSA in non-      All patients had mechanical heart valve prostheses: 15
                    O vs. O blood type was 1.37 (95% CI, 1.04 to 2.18); aOR        aortic valve prostheses (AVPs), 163 mitral prostheses
                                       on




                    of RSA in women with vs. without IT was 2.06 (95% CI,          (MVPs) and 75 combined MVPs and AVPs. Out of 307
                    1.08 to 3.61). Interestingly, we found concomitant IT and      pregnancies, there were 15 maternal deaths (4.9%), 24
                    non-O in n. 29 (13%) cases and n. 16 (7%) controls, aOR        thrombotic events (7.8%) and 22 major bleedings
                                  N




                    2.52 (95% CI, 1.12 to 5.47). Conclusions: We found a           (7.2%). The cause of death often is unknown.
                    significant higher prevalence of non-O blood group or IT       Thrombosis leading to death was more frequent than
                    in women with unexplained RSA. Their combined pres-            hemorrhage (33.3% vs. 6.7%). Out of 24 thrombotic
                    ence further increases RSA risk. Larger studies are war-       events, 23 were valve thrombosis. Major bleeding
                    ranted to validate our findings.                               occurred mainly after miscarriage/therapeutic abortion
                                                                                   compared to post-partum (12.9%, 12%, 3.9% respective-
                                                                                   ly; p=0.019). Fifty pregnancies (16.3%) were terminated
                    OC038                                                          by therapeutic abortion. Only 133/237 (56.1%) mothers
                                                                                   had at least one live baby. Nearly one out of five mothers
                                                                                   (19.8%) repeatedly tried to have a child, but only 41
                    PREGNANCY IN PATIENTS WITH MECHANICAL                          babies were born live after their collective 101 pregnan-
                    HEART VALVES: FOETO-MATERNAL OUTCOME IN                        cies. Information about VKAs embrio-foetopathy in our
                    THE POPULATION MONITORED BY                                    population was not available. Conclusions: To become a
                    ANTICOAGULATION SERVICE IN SALAM CENTRE                        mother is a natural desire all over the world. In medium-
                    FOR CARDIAC SURGERY-EMERGENCY                                  and low-income countries, the importance placed on
                    NGO – KHARTOUM – SUDAN                                         women having children, and the social pressure on them
                                                                                   to do so, is very strong. In this context, limitations to
                    N. Erba1,2,4, S. Abdelwahab2, S. Gatti1, M. Langer1,5,         family planning are not easily accepted, even if for para-




                                                                [Hematology Reports 2021; 13:s3]                                                [page 27]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 28



         Oral communications




                     mount medical reasons. In our population maternal mor-        rare inherited bleeding disorders. Their demographic
                     tality and major complications during pregnancy are at        profile, reason for admission to the ED, triage priority
                     an unacceptable level: this is related to the presence of     code, length of stay in the ED, time between triage and
                     the mechanical valve at first, but also to low compliance     the first replacement dose of lacking clotting factor, and
                     to anticoagulant therapy and scarcity of tertiary health-     outcome were investigated. Descriptive analysis is
                     care services. Safeguarding mothers’ lives must become        reported. Significance was set at p<0.05. Results:
                     our principal commitment, and solutions should be con-        Overall, 60 children were admitted 210 times (median
                     text-related. First, more efforts should be made to           age: 8.00 years): 41 with haemophilia A, 7 haemophilia
                     improve women’s awareness about health issues related         B, 9 von Willebrand disease, and 3 other rare bleeding
                     to pregnancy. Standardized counseling, focused on self-       inherited disorders (2 with factor VII deficiency, 1 with
                     engagement, can strengthen alliance between patients          factor XI deficiency). On the same period, 58 adults
                     and caregivers, improve adherence to therapy and con-         were admitted 115 times (median age: 38.00 years): 40
                     traception, and ensure timely communication about             with haemophilia A, 9 haemophilia B, 8 von Willebrand
                     pregnant status. At last but not at least, a nationwide       disease, and 1 factor V deficiency. Triage priority code
                     obstetric support network should be implemented to            was significantly higher in children, with red and yel-
                     meet this complex and growing health demand.                  low high priority codes attributed respectively to
                                                                                   190/210 (90.5%) admissions of children and 42/115
                                                                                   (36.5%) admissions of adults (p<0.001). Suspected
                     OC039                                                         spontaneous bleeding was reported in 70/210 pediatric




                                                                                                           ly
                                                                                   admissions (33.3%) and 59/115 adult admissions




                                                                                                    on
                     IMPACT OF TRIAGE PRIORITY CODE ON THE                         (51.3%). Median time between triage and the first
                                                                                   replacement dose of clotting factor was 49.00 minutes
                     MANAGEMENT OF CHILDREN AND ADULTS WITH
                                                                                   (IQR 23.00-85.00) in children and 157.00 minutes
                     INHERITED BLEEDING DISORDERS IN THE
                                                                                   (78.00-242.50) in adults (p <0.001). Compared to




                                                                                              e
                     EMERGENCY DEPARTMENT                                          adults, children showed significantly shorter median
                     C. Ferraris1, L. Franceschi1, E. Castagno2, B. Pollio3,
                     I. Ricca3, R. Albiani3, A. Borchiellini4, C. Dainese4,
                                                                                       us
                                                                                   time since triage to clinical examination (15 minutes vs
                                                                                   47 minutes, respectively; p<0.001), shorter median
                                                                                   length of stay in the ED (106 minutes vs 311 minutes,
                     A. Catalano5, R. Cerchio2, C. Bondone2
                                                                                   al
                                                                                   respectively; p<0.001), and lower prevalence of hospi-
                     1Department                                                   talization (8.1% vs 15.2%, respectively; p=0.049).
                                                                           ci

                                   of Public Health and Pediatrics, University     Conclusions: The upgrade of triage priority code for
                     of Turin, Turin, Italy; 2Department of Pediatric              children with inherited bleeding disorders was associat-
                                                                     er


                     Emergency, Regina Margherita Children’s Hospital -            ed to earlier administration of factor, shorter length of
                     A.O.UCittà della Salute e della Scienza di Torino, Turin,     stay in the ED, and lower hospitalization rate, which
                                                               m



                     Italy; 3Transfusion Medicine, Department of Diagnostic,       represent indirect indicators of better outcome. Our
                     Regional Center for hemorragic and thrombotic diseases        observation provides evidence supporting the recom-
                                                   om




                     in childhood, Regina Margherita Children’s Hospital -         mendations not to keep patients with inherited bleeding
                     A.O.UCittà della Salute e della Scienza di Torino, Turin,     disorders waiting in the ED and to provide early
                     Italy; 4Hematology Division, Regional Center for              replacement of lacking clotting factors.
                                                -c




                     hemorragic and thrombotic diseases, A.O.UCittà della
                     Salute e della Scienza di Torino, Turin, Italy; 5Department   References:
                                        on




                     of Clinical and Biological Sciences, University of Turin,     1 Srivastava A, Santagostino E, Dougall A, Kitchen S,
                     Orbassano (TO), Italy                                            Sutherland M, Pipe SW, et al. WFH Guidelines for
                                                                                      the Management of Hemophilia, 3rd edition.
                                   N




                     Background and Aims: Haemophilia and von                         Haemophilia 2020;26(Suppl 6):1-158.
                     Willebrand disease are the most common inherited              2 Dunkley S, Lam JCM, John MJ, Wong RSM, Tran
                     bleeding disorders in the Emergency Department (ED).             H, Yang R, et al. Principles of haemophilia care: The
                     Early replacement of the lacking clotting factor is rec-         Asia-Pacific perspective. Haemophilia 2018;24:
                     ognized as the cornerstone for better outcome, based on          366-75.
                     consensus of experts but lacking high-level evidence
                     [1,2]. For this reason, in most pediatric EDs children
                     with known inherited bleeding disorders usually receive
                                                                                   OC040
                     high triage priority code even for not serious events.
                     Our aim was to investigate the impact of triage priority
                     code on the management of children with inherited             LPA KRINGLE IV TYPE 2 DETECTION: QPCR OR
                     bleeding disorders admitted to ED and their outcome,          DDPCR?
                     compared to adults. Methods: All the children <18
                     years and adults with inherited bleeding disorders            G. Barbieri1, T. Capezzuoli1, M. Giannini1, G. Cassioli1,
                     referred to the Pediatric ED and General ED of our            F. Cesari2, R. Marcucci3, A.M. Gori3, B. Giusti3,
                     Institution between January 2015 and June 2021 were           E. Sticchi3
                     retrospectively reviewed. We included patients with
                     haemophilia A and B, von Willebrand disease and other         1University   of Florence, Department of Experimental




         [page 28]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 29



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    and Clinical Medicine, Florence; 2Atherothrombotic           tively). Conclusions: Data obtained by this study show
                    Diseases Center, Careggi Hospital, Florence;                 the limitations in terms of stability and sensitivity of
                    3University of Florence, Department of Experimental          qPCR, enhancing the more accurate and less variable
                    and Clinical Medicine, Atherothrombotic Diseases             results obtained with ddPCR, increasingly widespread
                    Center, Careggi Hospital, Florence, Italy                    method in diagnostics. The complex nature of this kind
                                                                                 of polymorphism still makes difficult to find an effec-
                    Background: Evidence to support the role of lipopro-         tive method to determine the exact number of repeats of
                    tein (a) [Lp(a)] as a risk factor for atherothrombosis       the KIV2 domain, nevertheless it is important to refine
                    continue to increase. Lp(a) levels strongly differ among     the techniques, since Lp(a) measurement has been
                    individuals, and many studies have shown the relation-       shown to provide clinically significant improved car-
                    ship between variants in the LPA gene, encoding              diovascular risk reclassification.
                    apolipoprotein (a), and the increase in Lp(a) levels,
                    closely related to the increase in cardiovascular risk.
                    The major genetic determinant of these levels is a copy      OC041
                    number variation (CNV) polymorphism, consisting of a
                    variable number of repeats of a 5 kb region that
                    includes exons 4 and 5 of the gene, encoding the protein     EFFECTS OF TRINITRO-RESVERATROL A NITRIC
                    domain Kringle IV type 2 (KIV2). Determination of            OXIDE-RELEASING DERIVATIVE OF
                    KIV2 repeat can be used to indirectly assess Lp(a) lev-      RESVERATROL, ON PLATELET ACTIVATION




                                                                                                        ly
                    els and to evaluate its involvement in modulating car-
                    diovascular risk. However, the peculiar structural char-     G. Guglielmini1, E. Berardi1, F. Messina2,




                                                                                                 on
                    acteristics of this variant constitute a significant chal-   M.C. Marcotullio2, P. Gresele1
                    lenge to the development of accurate methods for its
                                                                                 1Divisionof Internal and Cardiovascular Medicine,
                    detection. In this study, we compared quantitative real-




                                                                                             e
                    time PCR (qPCR) and digital droplet PCR (ddPCR) in           Department of Medicine and Surgery; 2Department of
                    KIV2 repeats determination. Methods: 70 patients with
                    probable/certain         diagnosis       of
                    Hypercholesterolemia according to the Dutch Lipid
                                                                     Familial       us
                                                                                 Pharmaceutical Sciences, University of Perugia, Italy

                                                                                 Background and Aims: Resveratrol (RSV), a
                                                                                 polyphenol of red wine, has been considered as a cru-
                                                                                 al
                    Clinic Network score were analysed. Demographic and
                    laboratory/clinical characteristics of the study popula-     cial contributor to the «French paradox» which
                                                                         ci

                    tion were collected. CNV values were obtained with           showed a protective effect of moderate wine consump-
                    qPCR using the 7900HT Sequence Detection System              tion against cardiovascular events. RSV stimulates
                                                                   er


                    and with ddPCR using the QX200 Droplet Generator             endothelial nitric oxide (NO) synthase by complex
                    and reader system. Telomerase reverse transcriptase          mechanisms thus enhancing NO production. In condi-
                                                             m



                    gene (TERT) was used as a single-copy reference gene         tions of high atherosclerotic risk, in which the antiox-
                    in both techniques. To make CNV values comparable            idant potential of plasma is strikingly impaired, the
                                                 om




                    within and between different plates, three internal con-     bioavailability of NO is reduced diminishing his car-
                    trols (CNV: 41<C1<47, C2=50, 61<C3<68) were used.            diovascular protective action, thus the use of NO-
                    Results: In the whole cohort, qPCR analysis showed           releasing compounds can have relevant therapeutic
                                                                                 effects. Aim of the present study was to evaluate
                                              -c




                    median values of repeats of 28.37 [IQR: 20.25-40.21],
                    while ddPCR of 10.55 [IQR: 9.44-12.26]. Correlation          whether a newly synthetized nitro derivative of RSV
                                       on




                    analysis between the two methods was slightly signifi-       may act as a NO-donor and display an antiplatelet
                    cant, because of the greater dispersion of data obtained     effect. Methods: 3,5,4’-tri-oxy resveratrol (TN-RSV)
                    by qPCR compared with ddPCR. Control sample C2,              was synthesized at the Department of Pharmaceutical
                                  N




                    used as a reference sample for qPCR and estimated to         Sciences of the University of Perugia. Trans-
                    have around 50 repeats, was confirmed to have a mean         Resveratrol (RSV) was obtained from Sigma Aldrich.
                    value of 54.85±1.11 with ddPCR analysis.                     Collagen-induced aggregation was assessed by LTA
                    Measurement of the other two internal controls (C1,          on washed platelets and followed for 5 min. At the end
                    C3) showed a wider data dispersion with qPCR vs              samples were centrifuged at 12,000xg for 30sec and
                    ddPCR (mean±SD: 36.99±11.25 vs 44.28±2.74 and                surpernatant stored at -80°C for subsequent NOx
                    92.78±25.20 vs 65.02±3.53, respectively). Spearman’s         assay. NOx were measured using a colorimetric non-
                    rho test showed an inverse proportional correlation          enzymatic assay (Griess Reaction Assay). Platelet
                    between Lp(a) levels of each patient and the CNV poly-       reactive oxygen species (ROS) production was meas-
                    morphism, as expected, but higher and significant when       ured by flow-cytometry using the fluorescent probe
                    evaluated with ddPCR despite qPCR (R=-0.393,                 H2DCFDA. Washed platelets were preincubated for
                    p<0.001 vs R=-0.198, p=0.044, respectively). Dividing        40 min with increasing concentrations (0.1 to 5µM) of
                    patients in two groups based on Lp(a) concentration          RSV, TN-RSV or their vehicles and stimulated with
                    (300 mg/L as cut off), a significant lower number of         collagen (50µg/µl, 3min). Platelet adhesion to colla-
                    repeats of the KIV2 domain emerged among patients            gen under flow conditions was assessed in a parallel
                    with greater levels of Lp(a) compared with the other         plate perfusion chamber. Citrated whole blood was
                    group in both methods but with strongly evidence with        preincubated with RSV, TN-RSV or their vehicles,
                    ddPCR than in qPCR (P<0.001 and P=0.024, respec-             perfused over a collagen-coated coverslip at high




                                                              [Hematology Reports 2021; 13:s3]                                              [page 29]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 30



         Oral communications




                     shear rate (3000sec-1) for 5 min, then coverslips were       inflammation has been proposed as a potential mecha-
                     stained and surface coverage measured (IMAGE J).             nism underlying this link, although its consequences
                     Results: RSV exerted a mild inhibitory effect on             on comorbidities are not well elucidated. We aimed to
                     platelet aggregation with a J-shaped dose-response           disentangle the relationship between depression,
                     curve and a maximal inhibitory effect at 0.5µM (-            inflammation and cardiovascular risk by investigating
                     33.9±4.9%). On the contrary, TN-RSV induced strong           the role of specific biomarkers, gender, and symptoms
                     inhibition of collagen-induced platelet aggregation,         domains in this link. Methods: We performed gender-
                     already significant at 0.1µM and maximal at 0.5µM (-         stratified multivariable Cox regressions of first hospi-
                     52.4±16.6%,p=0.02). NOx increased dose-dependent-            talization (median follow-up 7.28 years) or mortality
                     ly in the supernatant of collagen-stimulated platelets       for cardiovascular disease (CVD) and cerebrovascular
                     preincubated with RSV. With TN-RSV the release of            disease (CeVD) (median follow-up 8.24 years) vs
                     NOx was significantly higher than with RSV over the          depression severity, in an Italian population cohort
                     entire concentration range tested (at 0.1µM                  (N=13,191; age≥35 years; 49% men), adjusting for
                     RSV=+21.4±3.4% vs TN-RSV=+62.5±7.3%,p=0.03)                  sociodemographic, health and lifestyle factors. We
                     maintaining similar difference up to 5µM.                    tested potential mediation of C-reactive protein, gran-
                     Preincubation of citrated whole blood with RSV               ulocyte-to-lymphocyte ratio (GLR), platelet and white
                     decreased platelet adhesion to collagen. TN-RSV              blood cell counts (WBC). Depressive symptoms were
                     0.1µM also inhibited platelet adhesion, but more             assessed through an alternative validated version of
                     strongly and its effect was enhanced by increasing the       the Patient Health Questionnaire (PHQ9-6). To inves-




                                                                                                         ly
                     incubation time (at 40min RSV=-55.2%±10.3 vs TN-             tigate specific domains of depressive symptoms, we
                                                                                  derived latent variables tagging somatic and cognitive




                                                                                                  on
                     RSV=-83.9±2.1%, p=0.02). Collagen-induced platelet
                     ROS formation was reduced by RSV with a J-shaped             symptoms, through a polychoric factor analysis. First
                     curve, with a significant maximal inhibition at 0.5µM,       hospital admissions were recorded by direct linkage
                     while TN-RSV inhibited it more strikingly and dose-          with the Molise regional registry of hospital discharge




                                                                                            e
                     dependently (at 0.5µM RSV=-57.1±3.9% vs TN-                  records, updated to December 31st, 2015. Overall and
                     RSV=-66.1±1.8%%, p=0.04). Conclusions: We syn-
                     thesized a novel RSV derivative with the capacity to
                     release NO, property that adds to the beneficial activ-
                                                                                     us
                                                                                  cause-specific mortality was assessed through interro-
                                                                                  gation of the Italian mortality (ReNCaM) registry and
                                                                                  linkage with the Moli-sani database through unique
                                                                                  al
                     ities of resveratrol the ability to act as a direct source   identifier codes for each participant, updated to
                     of NO, with a synergistic inhibitory effect on platelet      December 31st, 2015. Results: In 6,689 women,
                                                                          ci

                     actation. The addition of a NO-donating moiety to            N=630, N=88 and N=48 events for CVD, CeVD hos-
                     RSV may enhance its effectiveness in disease condi-          pitalizations and CVD deaths occurred, respectively.
                                                                    er


                     tions of in vivo peroxidation and strong atherosclero-       In 6487 men, CVD, CeVD hospitalizations or CVD
                     sis-related inflammation, such as those found in high-       death events were 1,020, 138 and 93, respectively.
                                                              m



                     risk patients with acute coronary syndromes.                 Depression severity was similarly associated with
                                                                                  CVD hospitalization in both men and women (Table
                                                  om




                                                                                  1). On the contrary, CeVD hospitalizations and CVD
                     OC042                                                        mortality relative risk was higher only in men with
                                                                                  severe depression (Table 1). In multivariable models
                                               -c




                                                                                  reciprocally adjusting for both factors, somatic
                     DISENTANGLING THE ASSOCIATION BETWEEN                        depressive symptoms resulted associated with CVD
                     DEPRESSION AND CARDIOVASCULAR RISK:
                                        on




                                                                                  hospitalization or death, and cognitive symptoms with
                     FINDINGS FROM THE MOLI-SANI STUDY                            CeVD hospitalization events (Table 1). In the associa-
                                                                                  tion between depression severity and CVD hospital-
                                   N




                     F. Bracone1, A. Gialluisi1,2, S. Costanzo1,                  izations, an explanatory effect of the leukocyte bio-
                     F. Santonastaso2,3,4, A. Di Castelnuovo5, S. Orlandi1,       markers of inflammation were revealed, with WBC
                     S. Magnacca5, A. De Curtis1, C. Cerletti1, M.B. Donati1,     and GLR explaining 2.4% (P=0.01) and 2.0%
                     G. de Gaetano1, L. Iacoviello1,2, on behalf of the           (P=0.03) of the association. A mediation role for WBC
                     Moli-sani Study Investigators                                (13.7%, p=0.01) and GLR (10.4%, p=0.11) was also
                                                                                  detected for CVD mortality risk. C-reactive protein
                     1Department   of Epidemiology and Prevention, IRCCS          and platelet count showed negligible mediation roles.
                     NEUROMED, Pozzilli, Italy; 2EPIMED Research                  Conclusions: We further characterized the link
                     Center, Department of Medicine and Surgery, University       between depression and related CVD risks, revealing
                     of Insubria, Varese, Italy; 3Human Technopole, Viale         a prominent explanatory role of immune components
                     Rita Levi Montalcini 1, 20157Milan, Italy; 4European         of inflammation, in particular granulocytes and lym-
                     School of Molecular Medicine, University of Milan,           phocytes, and highlighting the importance of gender
                     20122Milan, Italy; 5Mediterranea Cardiocentro, Napoli,       and of somatic depressive symptoms in this link.
                     Italy                                                        These findings point to the existence of pathways
                                                                                  other than inflammation through which depression
                     Background and Aims: Major Depressive Disorder               may play a detrimental effect on cardiovascular sys-
                     (MDD) is a mental illness associated with chronic            tem, possibly related to gender-specificity and symp-
                     conditions like cardiovascular disease. Circulating          tom clusters.




         [page 30]                                         [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 31



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    OC043                                                       before CCP treatment, were significantly associated
                                                                                with mortality (fibrinogen b=0.709 p<0.01 and D-dimer
                                                                                b=0.065 p<0.05). Only four out of 334 patients (1.1%)
                    COVID-19 CONVALESCENT PLASMA IN PATIENTS
                                                                                treated with CCP developed adverse events (i.e. ana-
                    WITH SARS-COV2 INFECTION: FOCUS ON                          phylaxis/hypersensitivity Grade 1-2) with a resolution
                    IMMUNOSUPPRESSED OR NEOPLASTIC                              in a day after CCP infusion suspension. Conclusions:
                    PATIENTS                                                    CCP treatment led to an increase in antibody levels in
                                                                                recipients, especially in ICU patients with severe
                    A.M. Palladino1,2, C.J. Tartari1, L. Russo1,                COVID-19. The low incidence of adverse events sup-
                    C. Giaccherini1, S. Gamba1, S. Bolognini1, C. Ticozzi1,     ports the role for CCP administration, especially in
                    M. Marchetti1, O.C. Cretu1, F. Schieppati1, L. Barcella1,   patients with severe inflammation and low antibody
                    F. Zunino1, A. Falanga1,2                                   levels at admission.
                    1Division   of Immunohematology and Transfusion
                    Medicine, Papa Giovanni XXIII hospital, Bergamo;            OC044
                    2Medicine and Surgery, University of Milan-Bicocca,

                    Milan, Italy
                                                                                A PLATELET LIPIDOMICS SIGNATURE IN
                    Background and Aims: Patients treated with immuno-          PATIENTS WITH COVID-19




                                                                                                        ly
                    suppressive and corticosteroid drugs are at increased
                    risk for morbidity and mortality due to SARS-CoV2           E. Petito1, L. Goracci2, A. Di Veroli2, E. Falcinelli1,




                                                                                                 on
                    infection. Passive immunization therapy with convales-      C. Bencivenga2, E. Giglio1, C. Becattini1,
                    cent plasma has been successfully used in prior viral       E. De Robertis3, G. Vaudo4, P. Gresele1
                    pandemics. For these reasons, from the beginning of the




                                                                                            e
                                                                                1Division of Internal and Cardiovascular Medicine;
                    COVID-19 pandemic our hospital has been actively
                                                                                2Department
                    involved in collection of hyperimmune plasma from
                    convalescent patients and in the administration of the
                    same plasma to hospitalized patients for SARS-CoV2
                                                                                    us       of Chemistry, Biology and Biotechnology;
                                                                                3Division of Anaesthesia, Analgesia, and Intensive

                                                                                Care, Department of Medicine and Surgery, University
                                                                                of Perugia, Perugia, Italy; 4Unit of Internal Medicine,
                                                                                al
                    infection. In this setting, in a cohort of COVID-19 plas-
                    ma recipients, we aim to evaluate the association of        Terni University Hospital, Terni, Italy
                                                                        ci

                    convalescent plasma treatment with clinical and labora-
                    tory outcome. Methods: Adult patients with SARS-            Background and Aims: Besides respiratory system
                                                                  er


                    CoV2 infection diagnosed with moderate to severe            involvement, venous thromboembolism is a severe
                    acute respiratory distress syndrome were considered for     complications of COVID-19, largely due to the strong
                                                             m



                    COVID-19-convalescent plasma (CCP) treatment.               derangement of hemostasis, with platelets playing a
                    Blood samples were collected before and after 7 days        central role (PMID: 35027697). Great attention has
                                                 om




                    from CCP administration and tested for anti-SARS-           recently been devoted to lipid alterations in COVID-
                    CoV2 Nucleocapsid (anti-N) and Spike protein (anti-S)       19, because viruses reprogram cellular lipid metabo-
                    IgG antibodies, markers of inflammation [i.e. fibrino-      lism in order to fuel their replication. Lipidomics
                                                                                studies in COVID-19 patients have been performed
                                              -c




                    gen, reactive protein C (CRP), lactate dehydrogenase
                    (LDH), lymphocytes], hypercoagulability (i.e. D-            mainly in plasma and serum (PMID: 33571544). To
                                                                                the best of our knowledge, platelet lipidomics have
                                      on




                    dimer) and organ functionality (i.e. AST, ALT, creati-
                    nine, GFR). Results: From April 2020 to April 2022,         not been examined despite the central role played by
                    334 hospitalized COVID-19 patients [103M/231F, age          platelets in COVID-19 complications. Aim of our
                                  N




                    range 67 (41-86) years] were treated with CCP, (84 ICU      study was to preliminarily explore whether platelet
                    and 250 not-ICU patients). Among them, 125 were             lipidomics are altered in COVID-19 patients com-
                    patients under immunosuppressive therapy (68 with           pared to age- and sex-matched healthy subjects, ana-
                    cancer, 56 with diabetes or other comorbidities). In the    lyzing lipidomic profile of ultrapurified platelets.
                    overall group (n=334), treatment with CCP led to a sig-     Methods: Eight hospitalized COVID-19 patients and
                    nificant increase in anti-SARS-CoV2 antibodies, both        eight age- and sex-matched healthy controls were
                    anti-N [3.5 (0.06-7.74) vs 5.74 (0.25-7.95), p<0.001]       enrolled. Platelets were ultrapurified by negative
                    and anti-S [63.15 (7.13-397.15) vs 123 (34-400)             selection using Dynabeads™ CD45 from peripheral
                    p<0.001], especially in ICU patients. Inflammatory          venous blood collected in trisodium citrate 3.2%
                    parameters (i.e. CRP, LDH and ferritin) significantly       (0.109 M, 1/10 v/v) (PMID: 30408636). Platelet
                    decreased after CCP treatment in the overall cohort of      lipids were extracted by LC-MS/MS using an untar-
                    patients, including those under immunosuppressive           geted lipidomics approach (PMID: 25381612) and
                    therapies. In particular, anti-N SARS-CoV2 antibodies       analyzed by the Lipostar software (PMID:
                    titre positively correlated with platelet counts (r=0.127   28471643). Results: No significant differences in
                    p<0.05) and negatively correlated with ferritin levels      clinical and demographic characteristics were
                    (r=-0.505 p<0.01), also after adjustment for age and        observed between patients and controls, except for
                    gender. Interestingly, in the sub-group of neoplastic       neutrophil count, neutrophil to lymphocyte ratio
                    patients (N=68), D-dimer and fibrinogen, measured           (NLR) and lymphocyte count which were significant-




                                                             [Hematology Reports 2021; 13:s3]                                             [page 31]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 32



         Oral communications




                     ly higher or lower, respectively, in COVID-19                     Materno-Infantili e dell’Adulto, Modena; *COVID-
                     patients. The final profile included 341 lipids cover-            19HD Study Group: DAschieri, AUSL Piacenza;
                     ing five categories (sterols, sphingolipids, glyc-                LBrugioni, AOU Modena; EClini, AOU Modena;
                     erophospholipids, glycerolipids, fatty acyls) and                 MCodeluppi, Piacenza Hospital; DImberti, Piacenza
                     about 20 subclasses. A full separation of the                     Hospital; GDolci, AOU Modena; E.Franceschini, AOU
                     lipidomics profile of COVID-19 platelets from that of             Modena; C.Mussini, University of Modena and Reggio
                     healthy controls was observed (Figure 1A). In partic-             Emilia; GPinelli, AOU Modena; APietrangelo,
                     ular, a significant decrease of ether phospholipids               University of Modena and Reggio Emilia; LSarti, AOU
                     (Figure 1B-D) and dihydro-Ceramide (Figure 1E),                   Modena; MSilva, AUSL Piacenza, Italy
                     and increased levels of ganglioside GM3 (Figure 1F)
                     were observed in platelets from COVID-19 patients                 Background and Aims: COVID-19 is associated with
                     compared to healthy controls. Conclusions: Our                    a severe procoagulant status, leading to a substantial
                     study shows for the first time that platelets from                incidence of venous thromboembolism (VTE), despite
                     COVID-19 patients display a peculiar lipidomics sig-              standard prophylaxis with Low Molecular Weight
                     nature distinguishing them from healthy controls, and             Heparin (LMWH). Several Randomized Controlled
                     suggests that altered platelet lipid metabolism may               Trials (RCT) have addressed the issue of whether high-
                     play a role in viral spreading and in the thrombotic              er-than standard prophylactic doses of LMWH might
                     complications of COVID-19. In particular, ether                   improve the prognosis of COVID-19 patients. On these
                     lipids are synthesized in peroxisomes (PMID:                      ground we designed the COVID-19 trial (Eudract N.




                                                                                                             ly
                     28523433), and SARS-CoV-2 decreases the number                    2020-001972-13), aimed at comparing the efficacy and
                     and alters the structure of peroxisomes (PMID:                    safety of two doses of Enoxaparin in non-critically ill




                                                                                                      on
                     34010015), thus reshaping the host cell environment               COVID-19 pts. Methods: COVID-19 HD is a multi-
                     in a way more suitable for its replication, given that            center, randomized, controlled, open label, two arms
                     peroxisomes are signaling platforms for antiviral                 study aimed at assessing whether Enoxaparin at dose




                                                                                                 e
                     innate immunity (PMID: 34696946).                                 of 70 IU/kg twice daily is more effective than

                                                                                          us
                                                                                       Enoxaparin 4000 IU once day to prevent clinical wors-
                                                                                       ening, and similar in terms of major bleeding risk, in
                                                                                       COVID-19 adult hospitalized patients with pneumonia
                                                                                       al
                                                                                       and coagulopathy (D-dimer >4 times the upper level of
                                                                                       normal reference range or Sepsis-Induced
                                                                               ci

                                                                                       Coagulopathy (SIC) score >4), not requiring invasive
                                                                                       mechanical ventilation (IMV). The Primary Efficacy
                                                                        er


                                                                                       Endpoint was clinical worsening during hospital stay,
                                                                                       defined as the occurrence of death, acute Myocardial
                                                                  m



                                                                                       infarction, VTE, or need for escalating the oxygen sup-
                                                                                       port. This outcome was analyzed as a binary outcome
                                                     om




                                                                                       as well as a time-to-event one. Primary safety endpoint
                     Figure 1. Untargeted lipidomic analysis of platelet samples. A)   was major bleeding, defined according to ISTH. A
                     Partial Least Squares Discriminant Analysis (PLS-DA) scores       sample size of 300 pts was planned. The first patient
                                                  -c




                     plot based on the controls (gray circles)/COVID-19 (black         was enrolled on June 30th, 2020. Because of the low
                     squares) classification. B-F) Variation of the lipid classes in   accrual rate due to the changed epidemiological sce-
                     COVID-19 patients compared to healthy subjects: B) ether
                                         on




                                                                                       nario, an interim analysis was performed on September
                     phosphatidylcholine (PC), C) ether lysophosphatidylcholine
                                                                                       2021, after the randomization of 132 patients, with the
                     (LPC), D) ether phosphatidylethanolamine (PE), E) Dihydro-
                     Ceramide (Dihydro-Cer), F) ganglioside GM3.
                                                                                       agreement of the Data Safety Monitoring Board.
                                    N




                                                                                       Results: Of the 132 pts examined, 96 were males and
                                                                                       36 females; 54.8% had 0-1 comorbidities, and 45.2% 2
                                                                                       or more. The mean age was 59.9 yrs (SD 10.8), and
                     OC045
                                                                                       only 10 pts. were older than 75 yrs. The mean BMI was
                                                                                       30.6 (SD 5.0). The two groups were well balanced in
                     COVID-19 HD TRIAL: RESULTS OF THE INTERIM                         terms of demographic and clinical characteristics.The
                     ANALYSIS                                                          primary end-point occurred in 25.3% of pts. allocated
                                                                                       to the low dose and in 12.3% of those to the high dose
                     M. Marietta1, P. Mighali2, V. Coluccio1, F. Banchelli3,           arm (Relative Risk 0.48, 95% C.I. 0.22-1.04, P=0.055).
                     R. Vicini2, A. Gozzi2, P. Vandelli2, R. D’Amico3                  When the primary outcome was examined as a time-to-
                     on behalf of the COVID-19 Study Group*                            event one, the analysis of survival curves showed that
                                                                                       most of events occurred within 5 days from of hospi-
                     1Struttura Semplice Malattie della Coagulazione,                  talization. This correspond to an incidence rate of
                     Azienda Ospedaliero-Universitaria di Modena; 2Servizio            20.8% /pts/week in the low dose and of 8.9%/pts/week
                     Formazione, Ricerca e Innovazione, Azienda                        in the high dose, with an Incidence Rate Ratio of 0.43
                     Ospedaliero- Universitaria di Modena; 3Università di              (95% C.I 0.18-0.99, P=0.042). No major bleeding was
                     Modena e Reggio Emilia, Cattedra di Statistica Medica,            reported in either group. Discussion: In this analysis,
                     Dipartimento di Scienze Mediche e Chirurgiche                     we found that the use of Enoxaparin 70 IU/kg twice




         [page 32]                                             [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 33



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    daily was associated to a trend toward a lower risk of       acute COVID-19 infection and 46 healthy subjects (HS)
                    clinical worsening in non-critically ill hospitalized        were used for comparison. Results: Dispnea in LCS
                    COVID-19 pts. Moreover, the time-to event analysis           patients was confirmed by CPET showing compro-
                    showed a statistically significant lower incidence rate      mised alveolus-capillary membrane diffusion and resid-
                    of the primary outcome in the high-dose arm. Both            ual pulmonary impairment. TFpos-platelet and -MV lev-
                    treatments proved to be safe, with no major bleeding         els were 3- and 2-fold lower at 6mo-FU compared to
                    reported in either group. Although preliminary, these        acute phase, being comparable to HS, as well as pTGC.
                    findings are in keeping with those of the larger             At 6mo-FU, the MV profile (total number and derived
                    ATTACC,         ACTIV-4a,          and     REMAP-CAP         from different cells) returned to physiological levels.
                    Multiplatform RCTs. A limit of this study is the low         Conversely, although lower than that measured in acute
                    accrual rate, which lengthened randomization for sev-        phase, a 2.5-fold higher platelet P-selectin expression
                    eral months, thus including pts. infected by different       and PLA formation was observed at 6mo-FU compared
                    SARS-CoV-2 variants, and possibly with different             to HS. Interestingly, a significant correlation between
                    clinical characteristics of the disease. The data from the   PLA formation and residual pulmonary impairment was
                    COVID-19 HD will be included in two large ongoing            observed. Conclusions: These data strengthen the
                    metanalysis, whose results will hopefully provide a          hypothesis that the presence of PLA in the bloodstream,
                    better evidence about the efficacy and safety of high        and thus also in the pulmonary microcirculation, may
                    doses of LMWH in preventing clinical worsening in            contribute to support pulmonary dysfunction still
                    hospitalized, non-critically ill COVID-19 pts.               observed in LCS patients.




                                                                                                        ly
                                                                                                 on
                    OC046                                                        OC047




                                                                                           e
                    LONG COVID-19 SYNDROME: INSIGHTS INTO THE                    EFFICACY AND SAFETY OF DIFFERENT HEPARIN
                    CONTRIBUTION OF PLATELET ACTIVATION TO
                    PULMONARY IMPAIRMENT                                            us
                                                                                 REGIMENS FOR PREVENTION OF VENOUS
                                                                                 THROMBOEMBOLISM IN HOSPITALIZED
                                                                                 PATIENTS WITH COVID-19: A META-ANALYSIS
                    M. Conti1, P. Canzano1, M. Brambilla1, A. Becchetti1,
                                                                                 al
                    P.G. Agostoni1, M. Pengo2, E. Tortorici2, M.E. Mancini1,     M. Graziani, C. Vedovati
                    D. Andreini1, A. Bonomi1, G.F. Parati2, M. Camera1,3
                                                                         ci

                                                                                 Medicina Interna Vascolare-Stroke-Unit, Università
                                                                   er


                    1Centro   Cardiologico Monzino IRCCS, Milan, Italy;          degli Studi di Perugia, Perugia, Italy
                    2Istituto Auxologico Italiano, Milan, Italy; 3Department
                                                             m



                    of Pharmaceutical Sciences, Università degli Studi di        Background: Venous thromboembolism (VTE) is
                    Milano, Italy                                                common in patients with coronavirus disease-2019
                                                 om




                                                                                 (COVID-19). The optimal heparin regimen remains
                    Background and Aims: Long-COVID-19 syndrome                  unknown and should balance thromboembolic and
                    (LCS) is defined as symptoms persisting beyond initial       bleeding risks. The aim of this study is to evaluate the
                    phase of infection. Among them, pulmonary fibrotic
                                              -c




                                                                                 efficacy and safety of standard or higher heparin regi-
                    damage remains in 25-30% of COVID-19 patients at 3-          mens for the prevention of VTE in patients hospital-
                                       on




                    6 month-follow-up. We documented that acute COVID-           ized due to COVID-19. Materials and Methods: We
                    19 patients have massive platelet activation character-      performed a systematic literature search; studies
                    ized by the formation of platelet-leukocyte aggregates       reporting on hospitalized patients with COVID-19 who
                                  N




                    (PLA), that may be involved in the pulmonary                 received standard heparin prophylaxis vs. high (inter-
                    microthrombi found in autoptic specimens, and by a           mediate or therapeutic) heparin regimens were includ-
                    prothrombotic phenotype. No data are currently avail-        ed if outcome events were reported by treatment group
                    able on contribution of platelet activation to residual      and more than 10 patients were included. Primary
                    pulmonary impairment and procoagulant potential in           study outcome was in-hospital VTE. Secondary study
                    LCS patients. Thus, aims of this study was to character-     outcomes were major bleeding (MB), all-cause death,
                    ize platelet activation, microvesicle (MV) profile,          fatal bleeding and fatal pulmonary embolism. Results:
                    platelet thrombin generation capacity (pTGC) in LCS          Overall, 30 studies (10637 patients) were included.
                    patients at 6-month-follow-up (6mo-FU) compared to           Venous thromboembolic events occurred in 5.4% and
                    acute COVID-19 infection patients. Methods:                  in 8.3% of patients who received heparin prophylaxis
                    Twentyfour 6mo-FU COVID-19 patients with estab-              with at high-dose or standard-dose, respectively (RR
                    lished LCS defined according to their residual pul-          0.72, 95% CI 0.56-0.93, I2 52.2%). MB was signifi-
                    monary impairment assessed by Cardiopulmonary                cantly higher in patients who received high- compared
                    Exercise Testing (CPET) and 64-rows-CT scan evalua-          to the standard-dose (3.8% vs 2.1%, RR 1.89, 95% CI
                    tion were enrolled. Platelet activation (P-selectin,         1.41-2.53, I2 18.1%). Sub-analyses showed a slight
                    Tissue Factor [TF] and PLA) and MV profile were eval-        benefit associated with high-dose heparin in patients
                    uated by flow cytometry; pTGC by calibrated automat-         admitted to non-intensive care unit (ICU) but not in
                    ed thrombogram. Fortysix patients enrolled during            those to ICU. No significant differences were observed




                                                              [Hematology Reports 2021; 13:s3]                                              [page 33]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 34



         Oral communications




                     for mortality outcomes. Conclusions: Heparin prophy-        coronary syndrome, 2 strokes) and 28 cases (16.6%) of
                     laxis at high-dose reduces the risk of VTE, but             VTE, mostly distal and peri-catheter deep venous
                     increased the risk of MB compared to the standard-          thrombosis (28.6% each). 41 patients (24.4%) were
                     dose. No clinical benefit for heparin high-dose was         transferred to semi-ICU or ICU, while 15 subjects
                     observed for ICU setting, but its role in the non-ICU       (8.9%) died, mainly because of respiratory or multior-
                     deserves further evaluation.                                gan failure and septic shock. None of bleeding or
                                                                                 thrombotic events were associated with PLTs count or
                                                                                 function. 62 underwent PLTs study at discharge, show-
                     OC048                                                       ing a slightly increased of median PLTs count
                                                                                 (282x109/L [211-386]) and PLT aggregation still nor-
                                                                                 mal: ASPI 69 AUC [29-72], ADP 67 AUC [45-87) and
                     PLATELET AGGREGATION IN MILD COVID-19:                      TRAP 88 AUC [63-116]. Two patients (1.2%) died
                     A PERSPECTIVE SINGLE CENTER STUDY                           within 15 days post-discharge (for unkown reasons) and
                                                                                 9 (5.4%) had a re-hospitalization within 30 days (1 for
                     C. Simion, E. Campello, L. Spiezia, D. Tormene,             rectorrhagia, 1 for a proximal DVT of the leg). None of
                     F. Dalla Valle, C. Bulato, M. Fadin, S. Gavasso,            these events were associated with PLTs count or func-
                     C.M. Radu, G. Saggiorato, F. Sartorello, P. Zerbinati,      tion. 30 days after discharge, PLT count was within the
                     P. Simioni                                                  normal range (292x109/L [221-356]), while vWF anti-
                                                                                 gen (217% [157-245]) and FVIII (179.9% [122.9-




                                                                                                        ly
                     General Medicine and Thrombotic and Hemorrhagic             236.8]) still increased. Aggregometry test showed nor-
                     Diseases Unit, Department of Medicine – DIMED,              mal PLT function: ASPI 57 AUC [38-71], ADP 59 AUC




                                                                                                on
                     University-Hospital of Padova, Italy                        [46-72] and TRAP 85 AUC [74-95]. Among patients
                                                                                 who were retested post-discharge, neither arterial or
                     Background and Aims: COVID-19 is associated with            VTE nor bleedings occurred. Conclusions: According




                                                                                             e
                     a peculiar coagulopathy characterized by increased risk     to our findings, mild COVID-19 patients present with
                     of arterial and venous thromboembolism (VTE). Little
                     is known about the role of platelets (PLT) function in
                     the pathophysiology of COVID-coagulopathy: mild
                                                                                     us
                                                                                 normal PLTs count and reduced thrombin-driven
                                                                                 platelet aggregation. This latter was restored at dis-
                                                                                 charge. No clinical events were associated with PLTs
                     thrombocytopenia and PLT hyperactivity have been            function.
                                                                                 al
                     described, particularly in later stages; defects of PLT
                                                                         ci

                     functional responses were also reported. The aim of our
                     study is to evaluate PLT function in in-patients with
                                                                   er


                     mild COVID-19 at admission and post-discharge
                     through impedance aggregometry. Methods: We
                                                              m



                     enrolled all consecutive adults admitted to our Internal
                     Medicine Unit between November 2020 and May 2021
                                                  om




                     for mild COVID-19. Exclusion criteria were ongoing
                     antiplatelet therapies, severe thrombocytopenia
                     (<50x109/L) and previous semi-intensive or intensive
                     care unit (ICU) stay. Clinical characteristics, blood
                                               -c




                     chemistry, coagulation panel and Multiplate® aggre-
                     gometry were collected within 48h after admission, at
                                       on




                     discharge and 30-day post discharge. Clinical outcomes
                     were incidence of venous or arterial thrombosis, any
                                   N




                     bleeding complications, transfer to semi-intensive or
                     ICU, in-hospital mortality, 30-day post-discharge hos-
                     pitalization and mortality. Results: Out of 247 patients
                     admitted, 168 were enrolled (median age 77 years [IQR
                     64–84], F 56.4%). At admission, median PLTs count
                     was within normal range (240x109/L [151-258], n.v.
                     150-450x109/L), while levels of von Willebrand factor
                     (vWF) antigen (247% [225-373], n.v. 42-170%) and
                     FVIII (176.1% [135.7-227.9], n.v. 50-150%) were
                     increased. Using impedance aggregometry, we
                     observed normal PLTs aggregation: ASPI 53 AUC [29-
                     72] (n.v. 21-112), ADP 63 AUC ([42-84], n.v. 48-119)
                     and reduced thrombin-driven aggregation: TRAP 53
                     AUC ([29-72], n.v. 86-159) (Figure 1). During hospital-
                     ization, bleeding occurred in 15 (8.9%) patients, partic-
                     ularly 3 muscle hematomas and worsening anemia
                     requiring iron supplementation or transfusion. We
                     reported 3 cases (1.8%) of arterial thrombosis (1 acute     Figure 1.




         [page 34]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 35



                                                                           XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                                    1Dipartimento  di Medicina Interna, U.Odi Medicina
                    OC049
                                                                                    Interna, Ospedale Santa Chiara, Azienda Provinciale
                                                                                    per i Servizi Sanitari (APSS), Trento, Italy; 2U.O di
                    MONTELUKAST REPURPOSING IN COVID-19                             Medicina Interna, Ospedale Santa Maria del Carmine,
                    PATIENTS                                                        Azienda Provinciale per i Servizi Sanitari (APSS),
                                                                                    Rovereto, Italy; 3U.Odi Anestesia e Rianimazione,
                    M. Brambilla1, P. Canzano1, A. Becchetti1, M. Conti1,           Ospedale Santa Chiara, Azienda Provinciale per i
                    G.E. Rovati2, M. Camera1,2                                      Servizi Sanitari (APSS), Trento, Italy; 4U.Odi
                    1Centro
                                                                                    Pneumologia, Ospedale Santa Chiara, Azienda
                            Cardiologico Monzino IRCCS, Milan, Italy;               Provinciale per i Servizi Sanitari (APSS), Trento, Italy
                    2Department of Pharmaceutical Sciences, University of
                    Milan, Milan, Italy                                             Background and Aims: Venous thromboembolism
                                                                                    (VTE) is associated with acute SARS-CoV2 infec-
                    Background and Aims: Sustained platelet activation,             tion, especially in cases with severe interstitial pneu-
                    thrombosis, vascular damage, fibrotic response as well as       monia, and influences its short-term prognosis. Data
                    inflammatory overload are typical features of COVID-19          on long-term consequences and follow-up of SARS-
                    pathology. Common denominator in these processes are            CoV2 associated VTE are still lacking. Therefore, we
                    leukotrienes (LTs). Elevated levels of LTE4 have been           aims to investigate long term recurrences, resolution
                    detected in bronchoalveolar lavage of COVID-19                  rates and outcome of SARS-CoV2 associated VTE.




                                                                                                           ly
                    patients so that the use of LT receptor antagonists as a        Methods: We conducted a cohort study of all SARS-
                    potential therapeutic for COVID-19 patient treatment has        CoV2 patients consecutively admitted for severe




                                                                                                    on
                    been hypothesized. A first phase III randomized double-         pneumonia to the intensive and sub-intensive care
                    blind clinical trial testing montelukast in COVID-19            units at the Azienda Provinciale per i Servizi Sanitari,
                    patients has been indeed proposed. Thus, aim of this            from March 21st to May 4th 2020, who developed an




                                                                                              e
                    study was to investigate whether montelukast affects the        episode of VTE (either pulmonary embolism, PE, or
                    expression of the major markers of platelet activation
                    such as tissue factor (TF), P-selectin, as well as the for-
                    mation of platelet-leukocyte aggregates and microvesicle
                                                                                       us
                                                                                    deep vein thrombosis, DVT). During hospitalization,
                                                                                    all patients were screened for DVT and, when appro-
                                                                                    priate, for PE. After discharge, we followed patients
                    (MV) release observed in COVID-19 syndrome.
                                                                                    al
                                                                                    with confirmed VTE for thrombotic and haemorrhag-
                    Methods: Blood from healthy subjects (HS; n=4-6) was            ic events at three, six and 12 months. Thrombus reso-
                    plasma-depleted and reconstituted with plasma pools
                                                                            ci

                                                                                    lution and cardiac involvement were evaluated by
                    (n=3) from COVID-19 patients (4 patients/pool) or from          venous and cardiac ultrasound. The post-COVID19
                                                                     er


                    the same HS blood donors. To assess the effect of mon-          functional status scale (PCFS) was used to investigate
                    telukast on cell activation, blood from HS was preincu-         residual symptoms and dysfunction. Thrombophilia
                    bated for 30 minutes with the drug. Circulating cell-asso-
                                                               m



                                                                                    screening (including factor V Leiden, prothrombin
                    ciated TF expression, platelet activation markers and MV        G20210A mutation, antiphospholipid antibodies, pro-
                    release were analyzed by flow cytometry. Results:
                                                   om




                                                                                    tein S, protein C and antithrombin levels) was per-
                    Plasma from COVID-19 patients significantly increased           formed at least one month after termination of antico-
                    (4-fold) the number of TFpos- and P-selectinpos-                agulation. Descriptive results are presented by mean
                    platelets of HS recapitulating the platelet activation status
                                                -c




                                                                                    and standard deviation where appropriate. Results:
                    of COVID patients. Montelukast prevented platelet acti-         As shown in Figure 1, of 218 patients admitted for
                    vation induced by plasma from COVID-19 patient and it
                                        on




                                                                                    severe SARS-CoV2 pneumonia, 35 (16%) had VTE
                    reduced the formation of circulating monocyte- and gran-        (mean age 66±11 years, 27 males). Five patients died
                    ulocyte-platelet aggregates, decreasing the number of
                                                                                    during hospitalization (all males, mean age 68±11
                                   N




                    those TFpos by 4-times. Finally, it completely inhibited
                                                                                    years). Of the 30 remaining (mean age 65±12, Brescia
                    the release of TFpos circulating MVs, reducing by more
                                                                                    SCORE at admission 2.8±0.9, duration of hospitaliza-
                    than 2-times those derived from platelets. Conclusions:
                                                                                    tion 41±15 days), 13 (43%) had PE and 17 (57%)
                    Our data indicate that leukotrienes contribute to sustain
                                                                                    DVT, of which 13 (43%) had isolated calf thrombo-
                    platelet activation occurring in the COVID-19 patient,
                                                                                    sis. At 12 months, all patients had complete resolution
                    which can however be prevented by treatment with mon-
                                                                                    of the DVT, and none had signs of pulmonary hyper-
                    telukast. Until results from ongoing trials will be avail-
                                                                                    tension at the cardiac ultrasound. Thrombophilia
                    able, our data provide the molecular basis by which the
                                                                                    screening was performed in 16 patients, and showed
                    drug may be effective in the treatment of COVID-19.
                                                                                    negative results. Anticoagulant therapy (22/30 low
                                                                                    molecular weight heparin; 5/30 direct oral anticoagu-
                                                                                    lant, 1/30 fondaparinux) was stopped after three
                    OC050                                                           months in most patients with DVT (13/17, 76%) and
                                                                                    six months in most patients with PE (7/13 54%). Only
                    SARS-COV2 ASSOCIATED VENOUS                                     six patients continued anticoagulation for 12 months
                    THROMBOEMBOLISM - A LONG TERM FOLLOW-UP                         (6/20, 20%). After a mean follow-up of 14.8±1.2
                                                                                    months, 6 (20%) minor bleeding and no VTE recur-
                    A. Maino1, M. Landolfo1, S. Conci2, E. Vettorato2,              rence occurred. Seven patients (54%) reported no
                    C. Susanna2, S. Magnoni3, F. Boccafoglio4, D. Peterlana1        limitations or symptoms at the PCFS, and the remain-




                                                                [Hematology Reports 2021; 13:s3]                                               [page 35]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 36



         Oral communications




                     ing reported mild to moderate limitations.                  dict the diagnosis of PE as confirmed by a contrast-
                     Conclusions: We did not observe recurrences of              enhanced CT lung scan. Results: During the study
                     SARS-CoV2 associated VTE. Our preliminary data              period, 74 COVID-19 patients who had CT angiogra-
                     suggest that stopping anticoagulation after a standard      phy for PE clinical suspicion were included (mean age
                     treatment period of three (for DVT) or six (for PE)         68 years, male 64.9%). Thirteen patients (17.6%) had
                     months might be a reasonable approach in SARS-              PE confirmed at CT. No significant differences were
                     CoV2 related VTE. Nevertheless, the risk of bleeding        observed for comorbidities, antithrombotic treatment
                     might be not negligible.                                    and mortality between the two groups. D-dimer result-
                                                                                 ed significantly higher in patients with compared to
                                                                                 patients without PE. Poor discrimination was observed
                                                                                 for Wells and Geneva scores (AUC 0.596, 95% CI
                                                                                 0.413-0.779, and AUC 0.603, 95% CI 0.439-0.767,
                                                                                 respectively), without substantial differences adding
                                                                                 d-dimer at conventional cut-off (Table 1). The
                                                                                 IMPROVEDD score had the highest discriminative
                                                                                 power among CPRs for VTE (AUC 0.699, 95% CI
                                                                                 0.539-0.860). Scores’ performance improved by
                                                                                 increasing the D-dimer cut-off at level of 2000 ng/ml:
                                                                                 among diagnostic scores, Wells showed the best dis-




                                                                                                        ly
                                                                                 crimination (AUC 0.806, 95% CI 0.674-0.939, nega-
                                                                                 tive predictive value 97%); among CPRs for VTE the




                                                                                                 on
                                                                                 IMPROVEDD confirmed its accuracy (AUC 0.769,
                                                                                 95% CI 0.633-0.904, negative predictive value 94%).
                                                                                 Conclusions: The accuracy of the currently used diag-




                                                                                            e
                                                                                 nostic and predictive scores for PE or VTE in COVID-
                     Figure 1.
                                                                                     us
                                                                                 19 patients is poor. D-dimer improves the diagnostic
                                                                                 accuracy of these scores; most of all, it seems to allow
                                                                                 a diagnostic strategy with a high negative predictive
                                                                                 al
                     OC051                                                       value, so we can rule out a consistent part of the
                                                                                 patients with a low risk of PE.
                                                                         ci

                     DIAGNOSTIC STRATEGIES FOR PULMONARY                         Table 1.
                                                                   er


                     EMBOLISM IN PATIENTS HOSPITALIZED FOR
                     COVID-19: ROLE OF CLINICAL PREDICTION
                                                              m



                     RULES
                                                  om




                     M.C. Vedovati, F. Balducci, C. Becattini

                     Internal, Vascular and Emergency Medicine - Stroke
                     Unit, University of Perugia, Perugia, Italy
                                               -c




                     Background: Diagnostic strategies for pulmonary
                                       on




                     embolism (PE) in patients already facing respiratory
                     failure due to COVID-19 is challenging. The use of
                                                                                 OC052
                                   N




                     the conventional diagnostic algorithm and clinical pre-
                     diction rules (CPR) for PE is controversial in these
                     patents. Methods: We conducted a retrospective              IMPACT OF COVID-19 PANDEMIC ON
                     cohort study in patients with COVID-19 infection,           PULMONARY EMBOLISM CHARACTERISTICS:
                     with the aim to evaluate the accuracy of currently          A MULTICENTRIC 24-MONTH PERSPECTIVE
                     available CPRs to assess the risk for venous throm-         STUDY
                     boembolism (VTE) in medically ill patients and algo-
                     rithms for the diagnosis of PE. Consecutive patients        P. Sterpone1, I. Abatangelo2, A. Raza3, F. Piacentino4,
                     >18 years hospitalized at Santa Maria della                 L. Tofanelli5, W. Ageno6,7, F. Pavesi2, S. Birocchi2,
                     Misericordia Hospital (Perugia, Italy) from March 2th,      M.P. Donadini6,7, G.M. Podda2
                     2020, to September 29th, 2021 were included if they
                     had: 1) confirmed diagnosis of COVID-19 with a              1Internal Medicine Residency Program, School of
                     molecular testing; 2) chest CT angiography performed        Medicine, University of Insubria, Varese; 2Medicine II
                     for clinical suspicion of PE during the hospital stay.      Unit, ASST Santi Paolo e Carlo, Department of Health
                     The study outcome was the accuracy of currently             Sciences, University of Milan, Milan; 3School of
                     available CPRs for PE diagnosis (Wells and Geneva)          Medicine, University of Insubria, Varese; 4Department
                     and for VTE-risk stratification in medically ill patients   of Diagnostic and Interventional Radiology, Ospedale
                     (IMPROVE, IMPROVEDD and Padua score) to pre-                di Circolo, ASST Sette Laghi, Varese; 5Radiology Unit,




         [page 36]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 37



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    ASST Santi Paolo e Carlo, Milan; 6Research Center on         with a significantly lower burden in terms of severity
                    Thromboembolic Disorders and Antithrombotic                  indicators and clot extension as compared to non-
                    Therapies, University of Insubria, Varese; 7Thrombosis       COVID-19 patients. All these data together suggest a
                    and Heamostasis Center, Ospedale di Circolo, ASST            potential role of lung infection causing local pul-
                    Sette Laghi, Varese, Italy                                   monary artery thrombosis instead of typical VTE with
                                                                                 clot embolization. Further research is needed to better
                    Background: COVID-19 is associated with intense              understand PE in COVID-19 patients.
                    systemic inflammation and abnormal coagulation
                    profile leading to an increased incidence of venous
                    thromboembolic events (VTE). We reported that the            OC053
                    incidence of Pulmonary Embolism (PE) during
                    COVID-19 pandemic was significantly increased of
                                                                                 IMPACT OF ROTATIONAL
                    about 20% as compared to pre-COVID-19 pandemic.
                    Few data are available on PE characteristics in              THROMBOELASTOMETRY (ROTEM) ON CLINICAL
                    COVID-19 patients. Nevertheless, it was suggested            COURSE AND OUTCOMES OF COVID -19
                    that occlusions of pulmonary arteries observed in            CRITICALLY ILL PATIENTS: A RETROSPECTIVE
                    COVID-19 patients are caused by pulmonary thrombi            SINGLE CENTRE OBSERVATIONAL STUDY.
                    originating in loco, as a result of a thrombo-inflam-
                    matory process. Our study aimed to evaluate the clin-        M. Pazzi1, E. Gemmi1, D. Conti1, L.A. Zanofino1,




                                                                                                        ly
                    ical, laboratory, and radiological characteristics of PE     L. Tofani2, D. Prisco3, V. Pavoni1, R. Marcucci4
                    in patients with or without COVID-19 Methods: We




                                                                                                 on
                                                                                 1SOC   Anestesia e Rianimazione Ospedale Santa Maria
                    conducted an observational multicentric cross-sec-
                    tional study on consecutive patients diagnosed with          Annunziata, Azienda USL Toscana Centro; 2Department
                    PE at admission or during hospital stay at Azienda           of Statistics, Computer Science, Applications, University




                                                                                           e
                    Socio Sanitaria Territoriale (ASST) Sette Laghi              of Florence; 3Department of Experimental and Clinical
                    (Varese) and ASST Santi Paolo e Carlo (Milan) dur-
                    ing 24 months, from12 months before (pre-COVID
                    period) up to 12 months after (COVID period)
                                                                                    us
                                                                                 Medicine, University of Florence; 4Center for
                                                                                 Atherothrombotic disease, AOU Careggi, Dept of
                                                                                 Experimental and Clinical medicine, University of
                                                                                 Florence, Italy
                                                                                 al
                    February 21st2020, when the first Italian case of
                    COVID-19 was diagnosed. Patients’ clinical and lab-
                                                                         ci

                    oratory characteristics were collected. Thoracic             Background and Aims: Recently a close correlation
                    Computed Tomography images (TC) were revised                 between prothrombotic laboratory alterations and
                                                                   er


                    and data on the distribution and extent of PE were           COVID-19 severity has been suggested and increased
                    collected and quantified by means of the Qanadli             D-Dimer (DD) levels have been reported as a nega-
                                                             m



                    Index. Non-COVID-19 patients (ie those of the pre-           tive prognostic factor. Although Viscoelastic tests
                    COVID period and those of the COVID period who               (VETs) have been used to detect hypercoagulability
                                                 om




                    tested negative for SARS-CoV-2) were compared to             (characterized by acceleration in clot formation time
                    COVID-19 patients. Results: During the study period          (CFT) and increased maximum clot firmness (MCF),
                    772 patients were identified with acute PE (336 in the       their usefulness in clinical management of critically
                                              -c




                    PRE-COVID period and 436 in the post COVID peri-             ill COVID-19 patients has yet to be elucidated. Our
                    od), of whom 88 had COVID-19. In patients with PE            study aimed to identify the relationship between
                                       on




                    and COVID-19, male sex was more common than in               ROTEM profiles and adverse outcomes (mortality,
                    non-COVID-19 patients (52% vs 41%; p-value 0.01).            needing for Mechanical Ventilation (MV), Venous
                    Presence of risk factors for VTE was more frequent in        Thromboembolism (VTE), COVID-19 Associated
                                  N




                    patients without COVID-19: previous VTE and active           Coagulopathy (CAC)) in critically ill COVID-19
                    cancer were significantly less common in COVID-19            patients. Methods: 100 consecutive patients admitted
                    patients compared to non-Covid-19 patients (3.5% vs          to the intensive care unit (ICU) with severe COVID-
                    11.6% p 0.02; 4% vs 24.4% p<0.0001 respectively).            19 pneumonia, between October 1, 2020 May 31,
                    Interestingly, median Troponin T and nT pro BNP              2021 were enrolled in Florence, Santa Maria
                    levels were lower (15 vs 56 ng/L p-value 0.04 and 424        Annunziata Hospital. (Table 1). Results: We enrolled
                    vs 1501 pg/ml p-value 0.01 respectively) in COVID-           a population with a significantly higher prevalence of
                    19 patients. In COVID-19 patients the median value           male (80%), hypertension and diabetes. As concerns
                    of Qanadli Index was inferior (4 vs 7 p<0.0008), right       standard coagulation tests and ROTEM profile, a pro-
                    ventricular dilatation was less frequent ( 4.3 vs 11.5 %     thrombotic pattern was documented, with high DD
                    p 0.07) and distal obstruction was more common               (>6 VN) and fibrinogen levels. A modest increase of
                    compared to negative patients. Conclusions: In               MCF in EXTEM and FIBTEM was instead depicted
                    COVID-19 patients the classic risk factors for VTE           probably due to early administration of “intermedi-
                    are less frequent, possibly reflecting that COVID-19         ate” LMWH dosage before ICU admission in moder-
                    is itself a strong risk factor. The different characteris-   ate respiratory failure patients. Regarding ICU mor-
                    tics of COVID-19 associated PE suggest a difference          tality, statistically significant differences were found
                    in pathophysiological mechanisms. Furthermore, our           between DD values and platelet counts. ROTEM
                    study found a predominantly distal distribution of PE        analysis highlighted any hypercoagulability differ-




                                                              [Hematology Reports 2021; 13:s3]                                               [page 37]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 38



         Oral communications




                     ence but Maximum Lysis (ML) values resulted statis-        OC054
                     tically different in EXTEM and INTEM. (Table 1) A
                     sub-group analysis demonstrated hypofibrinolysis in
                     MV patients too. However, as concerns MV rate, the         RETROSPECTIVE SINGLE-CENTER ANALYSIS OF
                     univariate analysis showed a possible association          THE EFFECTS OF HEPARIN PROPHYLAXIS IN
                     with hypercapnia (OR 1.11, 1.03-1.2, p=0.0053) and         COVID-19 PATIENTS ADMITTED TO MEDICAL
                     leukocytosis (OR 1.12, 1.02-1.23, p=0.0181). Patients      WARDS: ANTI-THROMBOTIC OR ANTI-VIRAL?
                     with VTEs (n=20) showed higher DD values
                     (10732±1703 ng/ml vs. 3773±747 ng/ml,                      A. D’Angelo, L. Galli, P. Della Valle, C. Tresoldi,
                     p=0,010319); conversely no correlation was high-           M. Spessot, D. Palumbo, F. De Cobelli, D. Baccellieri,
                     lighted in terms of ROTEM hypercoagulability or            L. Apruzzi, M. Locatelli, M. Tresoldi, S. Calvisi,
                     hypofybrinolisis. No statistical difference was found      P. Scarpellini, M. Ripa, G. Martino, E. Bosi, L. Dagna,
                     between CAC (n=29) vs no CAC (n=71) patients.              A. Castagna, P. Rovere-Querini, F. Ciceri, on behalf of
                     Lastly, survival analysis allowed us to identify           COVID-BioB Study Group
                     hypofibrinolysis (ML EXTEM; HR 0,87, 0,81-0,95,
                     p=0,0011and INTEM; HR 0,9, 0,84-0,97,                      Scientific Institute Ospedale San Raffaele and Vita e
                     p=0,0083)as a possible risk factor. Conclusions: In        Salute University, Milano, Italy
                     our COVID-19 population, NOT-survivors had higher
                     DD values, lower platelet counts and a more pro-           Background and Aims: Mechanisms whereby thera-




                                                                                                        ly
                     nounced hypofibrinolytic ROTEM pattern than sur-           peutic-dose heparin appears beneficial in non-critical-
                     vivors. Hypofibrinolysis occurs more frequently in         ly ill, but detrimental in critically ill covid-19 patients




                                                                                                 on
                     patients requiring MV, although it does not result as a    are unclear. We retrospectively evaluated the effect of
                     possible risk factor for intubation. Similarly, hyperco-   LMWH prophylaxis on the composite endpoint of
                     agulability is a common ROTEM feature but it               symptomatic thromboembolism, transfer to ICU,




                                                                                           e
                     appears to increase neither VTE risk, intubation rate,     major and NMCR bleeding, and 28-day mortality of
                     or mortality probably due to early administration of
                     intermediate-dose LMWH. Therefore, we could
                     hypostasize that hypofibrinolysis, but not hypercoag-
                                                                                    us
                                                                                covid-19 patients hospitalized in medical wards of the
                                                                                San Raffaele Hospital during the first wave of the pan-
                                                                                demic. Secondary outcomes were the individual con-
                     ulability, resulted in worse outcomes in terms of mor-
                                                                                al
                                                                                tributors to the composite endpoint. Methods: Of 519
                     tality risk, MV and VTE. 29% of patients met CAC           patients with laboratory confirmed respiratory syn-
                                                                        ci

                     criteria but no significant difference in mortality, MV    drome coronavirus 2 (SARS-CoV-2) infection admit-
                     rate, VTE and ROTEM parameters was revealed.               ted to the medical wards of the hospital from March
                                                                  er


                     Further studies are highly recommended in order to         1st to April 30th 2020, 56 were on therapeutic antico-
                     define the usefulness of VETs as a tool for multipara-     agulation before infection, 243 received prophylactic
                                                             m



                     metric prognostic assessment. However, based on            anticoagulation, and 220 patients did not. A multivari-
                     current data, ROTEM analysis could represent a help-       able Cox proportional hazard model was used to
                                                 om




                     ful tool for predicting ICU admission risk and worst       assess the association between demographic and other
                     outcome as well as bleeding risk in critically ill         clinical or laboratory factors and the risk of ≥1 clinical
                     patients.                                                  event; the stabilized inverse probability weighting,
                                              -c




                                                                                calculated on propensity score for heparin prescrip-
                                                                                tion, was considered in the model. The hospital’s EC
                     Table 1.
                                       on




                                                                                approved the study, registered on ClinicalTrials.gov
                                                                                (NCT04318366). Results: Incidence rates of the com-
                                                                                posite endpoint (per 100 patient-days of follow up)
                                  N




                                                                                were 3.71 (2.54-5.41) for patients on therapeutic anti-
                                                                                coagulation, 3.30 (2.59-4.20) for those not receiving
                                                                                prophylaxis, and 1.99 (1.56-2.49) for those on prophy-
                                                                                lactic anticoagulation (p=0.003). In addition to latin
                                                                                ethnicity (HR 5.15), O2 saturation (HR per 1% higher
                                                                                0.93), lymphocyte count (HR per 1x109/L higher
                                                                                0.61), 4-items SOFA score (HR per 1 point higher
                                                                                1.32) and steroid use (any type and dosage, HR 0.54,
                                                                                all p values ≤0.03), all of the above at hospital admis-
                                                                                sion, heparin prophylaxis – considered as a time-vary-
                                                                                ing covariate (HR 0.38, 0.21-0.71, p=0.002) was pre-
                                                                                dictor of the 28-day risk of the composite endpoint’s
                                                                                occurrence. The incidence rate of 28-day mortality
                                                                                was the only individual contributor to the endpoint
                                                                                significantly different between patients receiving pro-
                                                                                phylaxis or not (0.49 vs 1.33 per 100 patient-days of
                                                                                follow up, p=0.0001). Notably, the incidence of major
                                                                                and CRNM bleeding, although numerically higher in




         [page 38]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 39



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    patients receiving prophylaxis, was not significantly       and 10 (39%) were taking Apixaban 2.5 mg BID for
                    different among the three groups. The results persisted     the prevention of VTE recurrence. The mean age was
                    in several sensitivity analyses. Conclusions: Similar       65±11.7 years, the mean BMI was 24.8±9.6, and the
                    to additional reports, in this retrospective analysis of    mean creatinine clearance was 78.9±32.9 ml/min
                    the early stage of the covid-19 pandemic, heparin pro-      (according to Cockroft–Gault). Regarding surgical
                    phylaxis reduced the 28-day mortality rate, but not the     interventions, 10 (38%) patients had undergone
                    rate of symptomatic thromboembolic events. The              ileostomy, 4 (15%) subtotal gastrectomy, 5 (19%)
                    SARS-CoV-2-specific anti-viral effect of heparin may        small bowel resection (mainly ileum), and 7 (27%)
                    explain why the covid-19 infection appears to be the        colon resection. Two patients had undergone both
                    first clinical setting showing a survival effect of         gastric resection and small bowel resection, while 2
                    heparin prophylaxis.                                        others both ileostomy and colectomy. The peak and
                                                                                trough plasma concentrations of patients on Apixaban
                                                                                5 BID for VTE treatment were respectively
                    OC055                                                       174.4±92.1 ng/mL and 72.25±41.3 ng/mL, on
                                                                                Apixaban 2.5 mg BID for VTE secondary prevention
                                                                                was 107.3±38.8 ng/mL and 50.4±14.2 ng/mL, and on
                    EFFECTS OF GASTROINTESTINAL RESECTION
                                                                                Apixaban 5 mg BID for NVAF was 304.7±183.1
                    AND OSTOMY SURGERY ON THE PLASMA
                                                                                ng/mL and 238.0±213.3 ng/mL [Figure 1]. We
                    CONCENTRATION OF APIXABAN:                                  observed that 2/26 (8%) patients, both receiving




                                                                                                         ly
                    AN OBSERVATIONAL PROSPECTIVE STUDY                          Apixaban 5 mg BID for the treatment of VTE, were




                                                                                                 on
                                                                                below the trough reference range. 1/26 (4%) patient,
                    R. Talerico1, R. Caprari1, A. Porfidia1, E. Fondi1,         on therapy with Apixaban 5 mg BID for the treatment
                    E. De Candia2, R. Pola1                                     of VTE, was below the peak reference range.
                                                                                Conclusions: These results show that almost all




                                                                                            e
                    1Dipartimento    di Medicina Interna, Fondazione            patients (92% for trough and 96% for peak plasma
                    Policlinico Universitario AGemelli IRCCS, Università
                    Cattolica del Sacro Cuore, Roma, Italy; 2Dipartimento
                    di Diagnostica per Immagini, Radioterapia Oncologica
                                                                                    us
                                                                                concentrations, respectively) who had undergone
                                                                                resections of the gastrointestinal tract, during
                                                                                Apixaban therapy achieved plasma drug concentra-
                    ed Ematologia, Fondazione Policlinico Universitario
                                                                                al
                                                                                tions within the reference range. Despite the limita-
                    AGemelli IRCCS, Università Cattolica del Sacro Cuore,       tions of our study, undoubtedly due to the small sam-
                                                                          ci

                    Roma, Italy                                                 ple size and variety of surgeries, these results are
                                                                                encouraging for further studies.
                                                                   er


                    Background and Aims: Patients who underwent gas-
                    trointestinal resection or ostomy surgery may have
                                                             m



                    altered absorption of orally administered drugs. This
                    could be an important clinical problem in subjects
                                                 om




                    taking direct oral anticoagulants (DOACs), for whom
                    an evaluation of effective anticoagulation is not war-
                    ranted. In this study, we evaluated whether therapeu-
                                              -c




                    tic plasmatic concentrations of Apixaban can be
                    found in this category of patients. Methods: We
                                       on




                    designed an observational prospective study and             Figure 1.
                    included patients with medical history of gastroin-
                    testinal resection or ostomy surgery who were on oral
                                  N




                    anticoagulation with Apixaban for the treatment or          OC056
                    prophylaxis of venous thromboembolism (VTE)
                    and/or for the prevention of cardioembolic stroke due
                    to nonvalvular atrial fibrillation (NVAF). We meas-         PREVALENCE AND RISK FACTORS FOR
                    ured the peak and the trough (2-3 and 12 hours after        PULMONARY EMBOLISM IN PATIENTS
                    drug intake respectively) plasma concentrations of          HOSPITALIZED IN INTERNAL MEDICINE UNITS
                    Apixaban using a chromatographic assay and com-             FOR ACUTE EXACERBATION OF CHRONIC
                    pared the levels obtained with the expected levels, as      OBSTRUCTIVE PULMONARY DISEASE:
                    established by the European Medicines Agency. We            A PROSPECTIVE MULTI-CENTER FADOI STUDY
                    included in the study patients who had undergone
                    gastric, small and/or large intestine resection up to the   I. Giarretta1, F. Dentali1,2, P. Gnerre3, F. Pomero4, R. Re5,
                    descending colon and/or ostomy surgery. We exclud-          N. Mumoli6, F. Pieralli7, P. Di Micco8, M. Bigossi9,
                    ed resection of sigmoid colon and/or rectum due to          A. Mazzone6, F. Colombo10, D. Manfellotto11
                    the poor absorption properties of these bowel tracts.
                    Results: 26 patients were enrolled, of whom 12              1Medicina  Generale 1, Medical Center, Ospedale di
                    (46%) were taking Apixaban 5 mg BID for the treat-          Circolo e Fondazione Macchi, ASST Sette Laghi, Varese,
                    ment of VTE, 4 (15%) were taking Apixaban 5 mg              Italy; 2Department of Medicine and Surgery, Insubria
                    BID for the prevention of cardioembolism in NVAF,           University, Varese, Italy; 3Internal Medicine, “San




                                                              [Hematology Reports 2021; 13:s3]                                                  [page 39]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 40



         Oral communications




                     Paolo” Hospital, Savona, Italy; 4Division of Internal
                     Medicine, Michele and Pietro Ferrero Hospital,
                     Verduno, Italy; 5Presidio Ospedaliero SAndrea, ASL VC,
                     Vercelli, Italy; 6Department of Internal Medicine, ASST
                     Ovest Milanese, Magenta (MI), Italy; 7Intermediate Care
                     Unit, Azienda Ospedaliero-Universitaria Careggi,
                     Florence, Italy; 8Department of Internal Medicine and
                     Emergency Room, Ospedale Buon Consiglio
                     Fatebenefratelli, Naples, Italy; 9Section of Internal
                     Medicine and Thromboembolic Diseases, Department of
                     Medicine, Fondazione Policlinico Universitario
                     AGemelli IRCCS, Rome, Italyand Division of Population
                     Health & Genomics, Pat McPherson Centre for
                     Pharmacogenetics & Pharmacogenomics, School of
                     Medicine, Ninewells Hospital & Medical School,                 Figure 1.
                     University of Dundee, Dundee, United Kingdom;
                     10Internal Medicine Ward, Ospedale Niguarda Ca’

                     Granda, Milan, Italy; 11Department of Internal                 OC057
                     Medicine, Fatebenefratelli Hospital, Rome, Italy




                                                                                                            ly
                     Background and Aims: The differential diagnosis                PREDICTION OF EARLY MAJOR BLEEDING IN




                                                                                                     on
                     between acute exacerbation of Chronic obstructive pul-         ACUTE PULMONARY EMBOLISM PATIENTS:
                     monary disease (ECOPD) and pulmonary embolism                  EXTERNAL VALIDATION OF PE-SARD SCORE
                     (PE) in patients admitted to Internal Medicine Units is




                                                                                                e
                     often challenging. Furthermore, PE may itself be the           L.A. Cimini, M.C. Vedovati, P. Quadalti, A. Vinci,
                     trigger or may complicate the clinical course of ECOPD.
                     The aim of this study was to evaluate the PE prevalence
                     and the associated risk factors in patients affected by
                                                                                        us
                                                                                    C. Becattini

                                                                                    Internal, Vascular, and Emergency Medicine - Stroke
                                                                                    al
                     COPD, hospitalized for acute worsening of respiratory          Unit, University of Perugia, Perugia, Italy
                     symptoms. Materials and Methods: This was a multi-
                                                                            ci

                     center prospective observational study, promoted by the        Background and Aims: In the acute phase of pulmonary
                     Italian Federation of Associations of Hospital Doctors         embolism (PE), anticoagulant treatment is necessary to
                                                                      er


                     on Internal Medicine (FADOI). The primary outcome of           reduce mortality. The most worrying consequence is the
                     the study was the incidence of PE among patients admit-        occurrence of major bleeding. Recently, the PE-Sard
                                                                m



                     ted for ECOPD within 90 days from hospitalization; the         score (Syncope, Anemia, Renal Dysfunction) was
                     secondary outcome was to identify potential risk factors       derived and internally validated to predict major bleeding
                                                   om




                     associated with PE. From January 2015 to November              events in patients with acute PE. We validated the pro-
                     2016, 521 consecutive patients with ECOPD were                 posed PE-SARD model, regardless of the severity of PE
                     enrolled in 30 Italian centers and evaluated for PE            and the anticoagulant treatment. Methods: From an
                                                -c




                     according to clinical suspicion. Results: The estimated        observational cohort including consecutive patients with
                     PE incidence was 9.8% (95% CI, 7.25-12.35%). Patients          acute PE, we performed a preliminary post hoc analysis.
                                        on




                     with PE did not differ from those without PE in terms of       The primary outcome was the occurrence of major bleed-
                     age, sex, or comorbidities. The multivariate analysis          ings at 30 days. Results: Overall, 376 patients were
                     demonstrated that clinical signs of deep vein thrombosis       admitted with acute pulmonary embolism in our hospital.
                                   N




                     (DVT) was associated with a 20-fold increased risk of          Of those, 12 were excluded due to the lack of baseline
                     PE (OR 20.60; 95% IC, 6.92-61.35). Recent prolonged            data. 364 patients were included in the analysis and fol-
                     bed rest (OR 2.39; 95% IC 1.10-5.20), arterial systolic        lowed for 30 days after PE diagnosis. The mean age was
                     blood pressure (ASBP) <130 mmHg (OR 2.11; 95% IC               69.7 (±15.7) and 51.6% were female. During the follow-
                     1.08-4.14), and hypocapnia (OR 2.88; 95% IC, 1.50-             up period, 18 major bleedings occurred (5%), of those 2
                     5.51) were also associated with PE (Figure 1). The sur-        were fatal (0.6%). Among the PE-SARD items, only ane-
                     vival analysis showed no influence of PE on hospital           mia resulted as a major bleeding predictor (p=0.004).
                     stay and mortality. The prevalence of PE in patients with      43.9% of patients were classified as at the low risk (160
                     0, 1, 2, 3 or 4 of the identified risk factors progressively   patients), 43.9% as at intermediate risk (160 patients),
                     increases from 3.1 % in patients without risk factors to       and 10.4% (39 patients). Observed bleeding rates
                     50.0% in subjects with 4 factors. Conclusions: This            increased with the risk group, from 2.5% in the low-risk
                     study demonstrated a high prevalence of PE in patient          group to 7.7% in the high-risk group. C-statistic (AUC)
                     hospitalized for ECOPD which was associated with pro-          was 0.622 (95% CI 0.497-0.746). Conclusions: In a real-
                     longed bedrest, clinical signs of DVT, SAP<130 mmHg            life cohort of patients with acute PE, the predictive value
                     and hypocapnia. Further studies are needed to test the         of the PE-SARD bleeding risk score is fairly good in
                     reliability and the efficacy of a rule out approach includ-    identifying patients at intermediate and high risk of
                     ing these variables in COPD patients admitted for sus-         bleeding in the acute phase of PE. The role of the PE-
                     pected acute exacerbation.                                     SARD score should be assessed in management studies.




         [page 40]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 41



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                                different drugs, for at least 3 to 6 months of treatment.
                                                                                Patients with VTE should receive a decision on the
                                                                                duration of AT that is often not easy to make. START2
                                                                                POST-VTE register study is aimed to investigate how
                                                                                Italian physicians deal with this issue; in particular,
                                                                                when they take a decision on duration of AT after a
                                                                                recent VTE episode, which decision is taken and why,
                                                                                and what happens during follow-up of the patients. The
                                                                                present analysis examined thromboembolic or bleeding
                                                                                complications, and deaths due to VTE occurring during
                                                                                two years of follow-up in the patients who had already
                                                                                received a decision on AT. Methods: The START
                                                                                POST-VTE register includes 1100 Italian patients with
                                                                                a recent VTE event recruited in 16 clinical centers.
                                                                                Adverse events occurring during follow up were
                                                                                recorded. Results: In the present analysis we describe
                                                                                the 472 patients who had already received a decision on
                                                                                duration of AT and have completed the 2 years of fol-
                                                                                low-up. In 59.3% of the cases the treatment was




                                                                                                       ly
                                                                                stopped and extended in 40.7%. Patients who extended




                                                                                                on
                                                                                anticoagulation were followed for 398 patients/years
                    Figure 1.
                                                                                (pt-yrs); during this period, overall 10 adverse events
                                                                                were recorded: 2 major bleeding (rate 0.5 pt-yrs) (1
                                                                                gastrointestinaI and 1 cerebral); none was fatal; 2




                                                                                          e
                    OC058                                                       CRNMB, 5 minor bleeding; 1 VTE recurrence (rate 0.5

                    START2-POST VTE REGISTRY: OUTCOMES AFTER
                    TWO YEARS OF FOLLOW-UP
                                                                                   us
                                                                                pt-yrs). One patient died for Covid disease; 2 patients
                                                                                are on indefinite treatment for onset of atrial fibrilla-
                                                                                tion. One patient was lost at follow up. In patients who
                                                                                al
                                                                                discontinued anticoagulation 40 recurrent events were
                    E. Antonucci1, L. Migliaccio1, B. Cosmi2, E. Bucherini3,    recorded: 38 VTE (13%) [16/38 proximal DVT
                                                                        ci

                    S. Birocchi4, V. Fregoni5, T. Lerede6, E. Montevecchi7,     (42,1%)]; 2 events involved the cerebral arteries. 1 gas-
                    R. Pancani8, R. Parisi9, A. Serrao10, G.M. Podda4,11,       trointestinal bleeding (non fatal) was recorded in a
                                                                  er


                    S. Testa12, D. Poli13, G. Palareti1                         patient who developed gastric cancer. Mean time of
                                                                                events from discontinuation of treatment was 279 ±203
                                                             m



                    1Fondazione Arianna Anticoagulazione,     Bologna, Italy;   (38 – 695) days; 57% of patients with recurrence were
                    2UO     Angiologia e Malattie della coagulazione,           males; median age 68 years. Conclusions: In patient
                                                 om




                    Dipartimento di specialità diagnostiche e Medicina          who continued treatment major bleeding rate was lower
                    sperimentale, IRCCS AOU S.Orsola-Malpighi, Bologna,         than those recorded in clinical trials. None event was
                    Italy; 3SS Aziendale di Angiologia Faenza AUSL              fatal. The rate of recurrent VTE in patients who stopped
                                              -c




                    Romagna, Ravenna, Italy; 4Unità di Medicina 2, ASST         anticoagulation was more frequent in male sex, what
                    Santi Paolo e Carlo, Milan, Italy; 5UOC Medicina            confirms available data on VTE recurrence.
                                      on




                    Generale, ASST Valtellina e Alto Lario, Sondalo (SO),
                    Italy; 6USC SIMT, Centro Emostasi e Trombosi,
                    Ospedale Papa Giovanni XXIII, Bergamo, Italy; 7UOC          OC059
                                  N




                    Angiologia Ospedale San Giovanni Apostolo,
                    Castelfranco Veneto, Treviso, Italy; 8UO Pneumologia,
                    Azienda Ospedaliero-Universitaria Pisana, Pisa, Italy;      SHORT AND LONG-TERM RISK OF VENOUS
                    9UOSD Ipertensione e Patologie Endocrine Metaboliche        THROMBOEMBOLISM IN PATIENTS WITH ATRIAL
                    Angiologiche, Ospedale SSGiovanni e Paolo, Venezia,         FIBRILLATION. A SYSTEMATIC REVIEW AND
                    Italy; 10Sezione Ematologia, Dipartimento di                METANALYSIS
                    Biotecnologie Cellulari ed Ematologia Azienda
                    Ospedaliero Universitaria Policlinico Umberto I, Roma,      T. Bucci1, G. Franchino2, G. Galardo1,
                    Italy; 11Dipartimento di Scienze della Salute, Università   P.R.J. Ames3,4, D. Menichelli2, A. Farcomeni5,
                    Degli Studi di Milano, Milan, Italy; 12Dipartimento di      P. Pignatelli2, D. Pastori2
                    Medicina di Laboratorio, Centro Emostasi e trombosi,
                                                                                1Department
                    ASST Cremona, Cremona, Italy; 13SOD Malattie                               of General Surgery, Surgical Specialties
                    Aterotrombotiche, Azienda Ospedaliero Universitaria-        and Organ Transplantation “Paride Stefanini,”
                    Careggi, Firenze, Italy                                     Sapienza University of Rome, Rome, Italy; 2Department
                                                                                of Clinical, Internal Medicine, Anaesthesiological and
                    Background and Aim: Patients with acute venous              Cardiovascular Sciences, Sapienza University of Rome,
                    thromboembolism (VTE) need immediate active anti-           Rome, Italy; 3Department of Haematology, Dumfries
                    coagulant treatment (AT), currently performed using         Royal Infirmary, Cargenbridge, Dumfries, UK;




                                                             [Hematology Reports 2021; 13:s3]                                               [page 41]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 42



         Oral communications




                     4Department of Clinical, Internal, Anesthesiological and    extended treatment with various anticoagulants
                     Cardiovascular Sciences, Sapienza University of Rome,       beyond six months from a VTE index episode in can-
                     Rome, Italy; 5Department of Economics and Finance,          cer patients, with a VTE recurrence rate of 4.0%,
                     University of Rome “Tor Vergata”, Rome, Italy               1.1%, 2.0% and 4,0%, respectively. A clinical trial,
                                                                                 named APICAT, which is currently ongoing, is aimed
                     Background: Atrial fibrillation (AF) carries an             to evaluate efficacy and safety of extended anticoagu-
                     increased risk of thromboembolism. While the rela-          lation with full- and low-dose apixaban in cancer
                     tionship between AF and arterial thromboembolism is         patients who had a previous VTE. In the present study,
                     well known, its association with venous thromboem-          we provide real-world data on the efficacy and safety
                     bolism (VTE) is still controversial. Methods: We per-       of extended anticoagulation with low-dose rivaroxa-
                     formed a systematic review of PubMed and EMBASE             ban in cancer patients, after completion of a full anti-
                     databases and metanalysis of observational study. The       coagulation regimen of at least 6 months for the treat-
                     studies were divided according to the time of VTE           ment of acute VTE. Methods: This is a prospective
                     onset after the AF diagnosis as follows: 1) up to 3         observational study performed at the “Section of
                     months, 2) up to 6 months, 3) >6 months. Pooled haz-        Internal Medicine and Thromboembolic Diseases” of
                     ard ratio (HR) was used as measure of association.          the A. Gemelli University Hospital of Rome, Italy.
                     Results: We included 11 observational studies: 6 retro-     Cancer patients who received rivaroxaban 10 mg once
                     spective and 5 prospective. A total of 689,092 AF           daily (od) for the secondary prevention of VTE were
                     patients were considered for the analysis. In the up to 3   enrolled. All patients had previously completed a




                                                                                                        ly
                     months group, AF was associated with a higher risk of       cycle of at least six months of full-dose anticoagula-
                                                                                 tion for the treatment of a VTE qualifying event. For




                                                                                                 on
                     VTE (HR:5.92, 95% CI: 2.36-14.85), DVT (HR:5.35,
                     95%CI:1.73-16.49) and PE (HR:4.92, 95%IC:1.76-              VTE qualifying event, we intended either deep vein
                     13.8). When we analysed the up to 6 months group, AF        thrombosis (DVT) of the lower limbs (both proximal
                     was still associated with a higher risk of VTE              and distal), pulmonary embolism (PE), or VTE in




                                                                                            e
                     (HR:3.76, 95%CI: 1.65-8.55), DVT (HR:1.75,                  unusual sites, including splanchnic/abdominal veins,
                     95%CI:1.43-2.14) and PE (HR:3.4, 95%CI:1.82-6.37).
                     However, after 6 months from the onset, AF was not
                     associated with a higher risk for VTE (HR:1.02,
                                                                                     us
                                                                                 cerebral veins, jugular veins, and deep veins of the
                                                                                 upper limbs, either in the presence or the absence of a
                                                                                 central venous catheter (CVC). The primary efficacy
                                                                                 endpoint was any type of objectively verified recur-
                                                                                 al
                     95%CI:0.97-1.08), DVT (HR:1.2, 95%CI:0.99-1.45)
                     and PE (HR:1.13, 95%CI:1.01-1.26). Conclusions: An          rent VTE. Primary safety endpoints were major bleed-
                                                                         ci

                     increased risk of VTE is evident in the first 3-6 months    ings or clinically relevant non-major bleedings
                     after AF diagnosis. The early initiation of anticoagula-    (CRNMB), defined according to the criteria of the
                                                                   er


                     tion in AF patients may reduce not only the risk of         International Society of Thrombosis and Haemostasis
                     stroke but also the risk of VTE.                            (ISTH). Results: To date (17 May 2022) the study has
                                                              m



                                                                                 included 106 patients, whose baseline characteristics
                                                                                 and information about the qualifying VTE event are
                                                  om




                                                                                 shown in the attached Table 1.
                     OC060
                                                                                 Table 1.
                     LOW DOSE RIVAROXABAN TO PREVENT
                                               -c




                     RECURRENCES OF VENOUS
                     THROMBOEMBOLISM IN CANCER PATIENTS:
                                       on




                     A REAL-WORLD EXPERIENCE
                                   N




                     P. Santini1, C. Mosoni1, E. Porceddu1, A. Porfidia1,
                     A. Lupascu1, P. Tondi2, R. Pola1
                     1Medicina   Interna e Patologie Tromboemboliche,
                     Fondazione Policlinico Universitario AGemelli IRCCS
                     Roma; 2Servizio di Angiologia, Fondazione Policlinico
                     Universitario AGemelli IRCCS Roma, Italy

                     Background and Aims: Patients with active cancer
                     are a high-risk population for venous thromboem-
                     bolism (VTE) and its recurrences. For this reason,
                     international guidelines suggest long-term anticoagu-
                     lation for secondary prophylaxis beyond 6 months in
                     cancer patients. However, this is a conditional recom-
                     mendation with low level of evidence since no data are
                     available on the optimal type and dose of anticoagu-
                     lant that should be used. Studies such as DalteCan,
                     TiCAT, Hokusai Cancer and Select-D have evaluated




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                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Upon completion of a therapeutic regimen of full anti-      stitution in the D’D3 domain. Methods: VWF:Ag and
                    coagulation for at least 6 months, all patients have ini-   VWF:GpIbR were measured by chemiluminescence
                    tiated treatment with rivaroxaban 10 mg od. To date,        assays, FVIII:C by chromogenic assay; the VWF-
                    they have been followed for a median time of 333 days       FVIII binding (VWF:VIIIB) and pro-peptide levels
                    (IQR 156-484). So far, we have observed 4 objectively       (VWF:pp) by ELISA assays; ADAMTS-13:activity by
                    verified VTE recurrences, with a rate of 3.8%.              FRET-based-assay; the VWF multimeric pattern
                    Interestingly, all 4 recurrences were proximal DVTs of      (mVWF) by SDS-agarose-gel electrophoresis; risto-
                    the lower limbs in patients whose qualifying previous       cetin-induced platelet aggregation (RIPA) by Born
                    event was a proximal DVT of the lower limbs. In 2           assay. Molecular modeling was performed by using I-
                    patients, the VTE recurrence developed in the presence      TASSER software. In vitro expression of VWF WT
                    of extrinsic vascular compression. In terms of safety,      and p.P1127S variants was obtained by HEK-293 cells,
                    we have observed no major bleedings (0.0%) and 3            while their interaction with LRP1 was analyzed by flu-
                    CRNMB (2.8%). Conclusions: The interim results of           orescence assay. The single-nucleotide variant (SNV)
                    this ongoing study provide preliminary evidence that        of ASGR2 subunit was detected by PCR. Results: The
                    extended treatment with rivaroxaban 10 mg may be a          p.P1127S variant was clinically associated with a mild
                    valid and safe therapeutic strategy to prevent CTE          VWD type 1 phenotype. Although desmopressin infu-
                    recurrences in cancer patients. Large-scale studies are     sion normalized the patient’s VWF levels, the protein
                    needed to confirm these data.                               clearance resulted enhanced (t1/2=6.7h) than in normal
                                                                                subjects (t1/2=12±0.7h). The VWF:pp/VWF:Ag ratio




                                                                                                      ly
                                                                                was slightly increased (1.3), reaching 1.52, after
                                                                                desmopressin infusion. The VWF:VIIIB and




                                                                                               on
                    OC061
                                                                                ADAMTS-13 activity were normal, as well as the
                                                                                RIPA assay and the mVWF pattern. The p.P1127S vari-
                    THE P.P1127S PATHOGENIC VARIANT LOWERS                      ant was normally synthesized and secreted by HEK-




                                                                                            e
                    VON WILLEBRAND FACTOR (VWF) LEVELS                          293 cells. Molecular modeling predicted the molecular
                    THROUGH HIGHER AFFINITY FOR THE
                    MACROPHAGIC SCAVENGER RECEPTOR LRP1:
                    CLINICAL PHENOTYPE AND PATHOGENIC
                                                                                    us
                                                                                structure of the VWF sequence 764-2191 for both the
                                                                                WT and p.P1127S variant. The presence of a Ser
                                                                                residue could perturb the non-covalent interactions
                                                                                between VWF monomers, causing a local disruption of
                                                                                al
                    MECHANISMS
                                                                                the correct folding in the Gly1115-Arg1133 region.
                                                                        ci

                    M. Tardugno, M. Sacco1, S. Lancellotti2,                    The loss of the hairpin-like structure produces long-
                    A. Branchini3, F. Bernardi3, F.M. Testa3, B. Lunghi3,       range allosteric effects, which propagate to the
                                                                  er


                    M. Pinotti3, B. Giusti4, G. Castaman5,                      Glu950-Ser1028 loop, globally generating a more
                    R. De Cristofaro1,2                                         opened conformation of the mature mutated VWF.
                                                             m



                                                                                These conformational changes were further investigat-
                    1Catholic  University of Sacred Heart; 2Fondazione          ed to predict their effects on the VWF and LRP1 inter-
                                                 om




                    Policlinico Universitario “AGemelli”, Istituto di           action. Compared to the WT, the molecular modeling
                    Ricovero e Cura a Carattere Scientifico (IRCCS);            of the p.P1127S variant showed an increased binding
                    3University of Ferrara; 4University of Florence;            affinity for the LRP1, confirmed by in vitro studies.
                    5Careggi University Hospital, Hemorrhagic and               Thus, the energetics, defined by ΔG, were -
                                              -c




                    Coagulation Disease, Department of Oncology, Italy          14.1Kcal/mol (Kd at 25°C=4.7x10-11M) and -
                                                                                15.2Kcal/mol (Kd at 25°C=7.3x10-12M) for the WT
                                      on




                    Background and Aims: Type 1 von Willebrand                  and p.P1127S variant, respectively (Figure 1). The
                    Disease (VWD) is extremely heterogeneous and vari-          index case was also homozygous for the rs2289645
                                  N




                    ably reduced VWF levels could depend on different           T>C in the ASGR2, potentially modulating VWF
                    molecular mechanisms, such as an alteration of secre-       clearance. Conclusions: Type 1 VWD mild phenotype
                    tion, intracellular retention and accelerated plasmatic     was induced by VWF p.P1127S mutation that causes
                    clearance. Moreover, a variety of cell surface recep-       functional changes in VWF conformation, increasing
                    tors, such as the low-density-lipoprotein receptor-relat-   the VWF affinity for the scavenger receptor LRP1,
                    ed protein 1 (LRP1) and the asialoglycoprotein recep-       thus accelerating the VWF clearance.
                    tor (ASGPR), binds with high affinity the FVIII/VWF
                    complex, modulating its half-life in circulation. Thus,
                    studies on rare missense variants could contribute to
                    better characterize VWD type 1 phenotypes. On this
                    background, this study aims to genetically and bio-
                    chemically characterize the molecular pathogenesis of
                    a mild haemorrhagic tendency in a 21-year-old Italian
                    woman with low levels of both VWF (VWF:Ag=34.3
                    IU/dL; VWF:GpIbR=32.8 IU/dL) and Factor VIII
                    (FVIII:C=55.3 IU/dL). The index case inherited from
                    her mother a new VWF heterozygous missense muta-
                    tion c.C3379>T in exon 25, causing the p.P1127S sub-        Figure 1.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 44



         Oral communications




                     OC062                                                      cases with and without enhanced clearance.
                                                                                Conclusions: Low-VWF levels can be associated with
                                                                                significant bleeding and >42% of patients had an
                     CLINICAL AND PHENOTYPIC PRESENTATION OF                    abnormal BS. Therefore, these patients should be con-
                     PATIENTS WITH LOW VWF IN THE MILAN CENTER                  sidered to have a mild bleeding disorder rather than a
                                                                                risk factor for bleeding. We further demonstrated the
                     O. Seidizadeh1,2, A. Ciavarella1, L. Baronciani1,          important role of VWF increased clearance as a patho-
                     M.T. Pagliari1, G. Cozzi1, P. Colpani1, S.M. Siboni1,      genic mechanism.
                     E. Biguzzi1, F. Peyvandi1,2
                     1Fondazione   IRCCS Ca’Granda Ospedale Maggiore            OC063
                     Policlinico, Angelo Bianchi Bonomi Hemophilia and
                     Thrombosis Center, Milan, Italy; 2Department of
                     Pathophysiology and Transplantation, Università degli      FINAL ANALYSIS FROM THE PIVOTAL PHASE 3
                     Studi di Milano, Milan, Italy                              HOPE-B GENE THERAPY TRIAL: STABLE
                                                                                STEADY-STATE EFFICACY AND SAFETY OF
                     Background: Low von Willebrand factor (VWF) refers         ETRANACOGENE DEZAPARVOVEC IN ADULTS
                     to patients with VWF levels of 30-50IU/dL. Managing        WITH SEVERE OR MODERATELY SEVERE
                     patients with low-VWF levels is a clinical challenge.      HEMOPHILIA B




                                                                                                         ly
                     Aim: To determine the clinical and phenotypic features
                     of patients with low-VWF. Methods: We included 235         W. Miesbach1, F.W.G. Leebeek2, M. Recht3, N.S. Key4,




                                                                                                 on
                     well-characterized patients with low-VWF levels            S. Lattimore3, G. Castaman5, M. Coppens6, D. Cooper7,
                     (VWF antigen and/or VWF activity 30-50 IU/dL with a        S. Slawka7, S. Verweij7, R. Gut7, R. Dolmetsch7, Y. Li8,
                     VWF activity/VWF antigen ratio>0.6). The ISTH              P.E. Monahan8, S.W. Pipe9, HOPE-B Investigators




                                                                                           e
                     bleeding assessment tool was used to assess the severity
                     and frequency of clinical symptoms. The VWF propep-
                     tide (VWFpp) assay was performed to determine VWF
                     clearance, with results compared to 120 healthy con-
                                                                                    us
                                                                                1University
                                                                                2Erasmus
                                                                                            Hospital Frankfurt, Frankfurt, Germany;
                                                                                          MC, University Medical Center Rotterdam,
                                                                                Netherlands; 3Oregon Health & Science University,
                     trols. The Mann-Whitney U test was used to compare         Portland, OR, United States; 4University of North
                                                                                al
                     medians between two independent groups. The abnor-         Carolina, Chapel Hill, NC, United States; 5Center for
                                                                        ci

                     mal bleeding score (BS) was defined as≥4 in adult          Bleeding Disorders and Coagulation, Careggi
                     males, ≥6 in adult females, and≥3 in children. Result:     University Hospital, Florence, Italy; 6Amsterdam
                                                                   er


                     The median (range) of VWF antigen and activity was         University Medical Centers, University of Amsterdam,
                     52 IU/dL(30-72) and 40 IU/dL(30-55) respectively,          Amsterdam, Netherlands; 7uniQure BV, Amsterdam,
                                                             m



                     with a VWF activity/VWF antigen ratio of 0.77. The         Netherlands/ uniQure IncLexington, MA, USA; 8CSL
                     median BS was 4 (n=160, range 0-17) and 33% of             Behring, King of Prussia, PA, United States; 9University
                                                  om




                     patients had bleeding that required treatment. Abnormal    of Michigan, Ann Arbor, MI, United States
                     bleeding was seen in 33% of children, 43% of females
                     and 51% of males. Epistaxis, bruising, surgery, menor-     Introduction: Etranacogene dezaparvovec (formerly
                                               -c




                     rhagia and bleeding from minor wounds were the most        AMT-061), an investigational gene therapy for hemo-
                     common clinical manifestations (Figure 1).                 philia B (HB), comprises an adeno-associated virus
                                       on




                                                                                serotype 5 (AAV5) vector, containing a codon-opti-
                                                                                mized Padua variant human factor IX (FIX) transgene
                                                                                under the control of a liver-specific promoter. Methods:
                                   N




                                                                                Adult male participants (n=54) with severe or moderate-
                                                                                ly severe HB (FIX ≤2%), with (n=33) or without (n=21)
                                                                                pre-existing AAV5 neutralizing antibodies (NAbs), were
                                                                                treated in the Phase 3, open-label, single-dose, single-
                                                                                arm, HOPE-B trial (NCT03569891) with 2x1013 gc/kg
                                                                                etranacogene dezaparvovec, following a ≥6-month lead-
                                                                                in period of FIX prophylaxis. FIX activity, annualized
                                                                                bleed rate (ABR), and FIX infusions were assessed over
                                                                                a uniform ≥52-week period after achieving stable FIX
                     Figure 1.                                                  expression, designated as Months 7–18 after vector infu-
                                                                                sion to serve as the basis for regulatory approval.
                     Nevertheless, severe bleeding symptoms as GI bleed-        Adverse events (AEs) were recorded. Results: Mean
                     ing and haemarthrosis were also observed. Patients         FIX activity was 39.0 IU/dL (±18.7; 8.2, 97.1) (standard
                     VWFpp/VWF antigen ratio was 1.4 (n=130, range 0.5-         deviation; min, max) at Month 6 and 36.9 IU/dL (±21.4;
                     2.4), significantly higher than that of controls (median   4.5, 122.9) at Month 18. Individual participant FIX
                     0.98, range 0.6-1.6, P<.002). An increased clearance       activity and titer of pre-existing AAV5 NAbs (present in
                     was observed in 35% of cases (n=46) evaluated for          21 participants) did not correlate up to a titer of 678; one
                     VWFpp. No difference was found for the BS between          participant with NAbs=3212 did not express FIX Padua.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 45



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Compared with the ≥6-month lead-in period (ABR               bearing the recurrent F8 PTCs (n=12, patient number
                    4.19), 52-week adjusted ABR for all bleeds during            from 10 to 57) arising from the highly frequent
                    Months 7–18 (ABR 1.51) was reduced by 64%                    CGA(arginine)>TGA change (p.R15X, p.R355X,
                    (p=0.0002), demonstrating statistical superiority over       p.R446X, p.R602X, p.R814X, p.R1715X, R1960X,
                    FIX prophylaxis. FIX-treated bleeds were reduced by          p.R1985X, p.R2135X, p.R2166X, p.R2228X and
                    77% (from ABR 3.65 to 0.83; p<0.0001). 52/53 (98%)           p.R2326X) were conducted in HEK293T cell lines to
                    participants who received a full dose discontinued pro-      detect luciferase activity, directly related to the amount of
                    phylaxis, with an overall 97% reduction in mean unad-        secreted full-length FVIII levels. Noticeably, the selected
                    justed annualized FIX consumption (from lead-in period       mutations are representative of 475/1053 (45%) HA
                    257338.8 to Months 13–18 8486.6 IU/yr/participant).          patients with nonsense mutations. All expressed F8 vari-
                    After dosing, 37 participants experienced 92 treatment-      ants showed FVIII traces due to spontaneous suppression
                    related AEs, of which 74 (80.4%) were mild. A serious        (“readthrough”), predicted to produce full-length pro-
                    AE of hepatocellular carcinoma was determined by             teins by inserting amino acid subsets including the origi-
                    independent molecular tumor characterization and vec-        nal residue at low frequency. Nevertheless, higher degree
                    tor integration analysis to be unrelated to vector.          of suppression (range 8-77% of wild-type FVIII-GL)
                    Conclusions: Following a single dose of etranacogene         was observed after ACE-tRNAArg treatment, which
                    dezaparvovec, participants experienced a stable and          restored secreted levels of wild-type FVIII-GL. In partic-
                    durable increase in mean FIX activity into the near-nor-     ular, the differential degree of correction allowed to strat-
                    mal range at 18 months and hemostatic protection,            ify F8 PTCs in three groups, namely i) low (p.R602X; 0-




                                                                                                          ly
                    achieving the HOPE-B primary efficacy endpoint.              10%), ii) intermediate (p.R15X, p.R446X, p.R814X,




                                                                                                  on
                                                                                 p.R1715X, p.R1985X, p.R2135X, p.R2166X; 10-50%,
                                                                                 and iii) high (p.R355X, p.R1960X, p.R2228X,
                    OC064                                                        p.R2326X; >50% WT) responders to ACE-tRNAArg-
                                                                                 mediated suppression. Noticeably, comparison between




                                                                                             e
                    ENGINEERED SUPPRESSOR TRNAS AS A NOVEL                       groups showed significantly higher secreted luciferase
                    CORRECTION APPROACH FOR RECURRENT
                    HEMOPHILIA A-CAUSING NONSENSE MUTATIONS
                                                                                     us
                                                                                 activity (p=0.0002) for the ACE-tRNA treated group.
                                                                                 Intrigued by these promising results in a system taking
                                                                                 advantage of a reporter gene, the efficiency of ACE-
                                                                                 al
                                                                                 tRNAArg was further challenged by inserting F8 PTCs
                    M.F. Testa1, J.D. Lueck2, A. Di Fiore1, F. Bernardi1,        in the proper FVIII-coding context. Preliminary results
                    M. Pinotti1, A. Branchini1
                                                                         ci

                                                                                 on the FVIII variants revealed higher secreted FVIII lev-
                                                                                 els after ACE-tRNAArg treatment, supporting the
                                                                   er


                    1Department   of Life Sciences and Biotechnology and         restored biosynthesis and secretion of wild-type FVIII.
                    LTTA Center, University of Ferrara, Ferrara, Italy;          Functional assays to assess activity levels are in progress.
                                                             m



                    2Department of Physiology and Pharmacology,
                                                                                 Conclusions: Our data propose a novel correction
                    University of Rochester School of Medicine and               approach for nonsense mutations, to faithfully introduce
                                                 om




                    Dentistry, Rochester, USA                                    the original amino acid and restore biosynthesis and
                                                                                 secretion of functional wild-type FVIII. This experimen-
                    Background and Aims: F8 nonsense mutations, rela-            tal evidence, provided for the first time in the coagulation
                    tively frequent cause (>10%) of Haemophilia A (HA),
                                              -c




                                                                                 field, opens the way for an innovative therapeutic
                    lead to the synthesis of truncated/loss-of-function factor   approach which might be translated to other diseases
                                       on




                    VIII (FVIII) variants by inserting premature termination     caused by nonsense mutations.
                    codons (PTCs), and are commonly considered “null
                    mutations”. Anticodon Engineered transfer RNAs (ACE-
                                  N




                    tRNA) are small molecules tailored to accurately drive
                                                                                 OC065
                    the insertion of the original amino at PTC during protein
                    synthesis, thus restoring production of the wild-type mol-
                    ecule. In this view, ACE-tRNAs might represent a novel       OSTEOCLAST ROLE IN HAEMOPHILIC BONE
                    correction approach to specifically suppress disease-        DISEASE
                    causing PTCs. In this pioneer project we challenged
                    ACE-tRNA to productively suppress highly frequent F8         M. Sacco1, S. Lancellotti2, G. Battafarano3,
                    PTCs and restore FVIII biosynthesis and secretion of         M. Tardugno1, L. Di Gennaro2, R. De Cristofaro1,2,
                    functional FVIII, with potential therapeutic implications.   A. Del Fattore3
                    Methods: Transient expression studies in HEK293T
                    cells through an optimized expression platform taking        1Catholic  University, Translational Medicine and
                    advantage of a fusion protein (FVIII-GL) joining FVIII       Surgery Department Rome; 2Fondazione Policlinico
                    and the naturally-secreted Gaussia luciferase (GL).          Universitario AGemelli, IRCCS, Center for
                    Treatment of cells with the ACE-tRNA engineered to           Hemorrhagic and Thrombotic Diseases and
                    introduce arginine (ACE-tRNAArg) on TGA F8 PTCs.             Haemophilia Center, Rome; 3Bone Physiopathology
                    Luciferase assays (FVIII-GL) and ELISA (FVIII) on cell       Group, Multifactorial Disease and Complex Phenotype
                    media to detect FVIII secreted levels. Results:              Research Area, Bambino Gesù Children’s Hospital,
                    Expression studies with the FVIII-GL fusion protein          Roma, Italy




                                                              [Hematology Reports 2021; 13:s3]                                                   [page 45]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 46



         Oral communications




                     Background and Aims: Bone disease is a relevant com-           OC066
                     plication of haemophilia A (HA), but its pathogenesis
                     still remains unknown. In people with hemophilia
                     (PWH), an association between hemophilia and low               NEW GENOME EDITING APPROACHES:
                     bone mineral density (BMD) has been found. Indeed,             BASE AND PRIME EDITING TO REVERT
                     43% of HA patients show low bone density and 27%               HEMOPHILIA A-CAUSING POINT MUTATIONS
                     develop osteoporosis. Over the last few years, clinical
                     and experimental evidences indicated that the deficiency       E. Tonetto1, S. Pignani2, G. Roman2, A. Follenzi2,
                     of FVIII leads to decreased BMD independently of               F. Bernardi1, D. Liu3, M. Pinotti1, D. Balestra1
                     hemarthrosis and medical comorbidities, suggesting a
                                                                                    1Department of Life Sciences and Biotechnology,
                     novel effect for FVIII outside of the coagulation system.
                     The reduction of BMD seems to be caused by perturba-           University of Ferrara, Ferrara, Italy; 2Department of
                     tions of the Receptor Activator of Nuclear factor-B            Health Sciences, University of Piemonte Orientale,
                     RANK Ligand (RANKL) and osteoprotegerin (OPG)                  Novara, Italy; 3Howard Hughes Medical Institute,
                     pathways. In haemophilic alterations of bone remodel-          Harvard University, Cambridge, USA
                     ing, osteoclasts seem to play an important role.
                     Osteoclasts are multinucleated cells which derived from        Background and Aims: Hemophilia A (HA), the most
                     the CD14+ monocyte/macrophage lineage. Until recent-           common coagulation bleeding disorder (1:5000 males),
                     ly, the identity of osteoclast progenitors has not been        is an X-linked hereditary disease caused by F8 gene




                                                                                                            ly
                     well defined, but evidences report that CD16−CD14+             mutation leading to deficiency of factor VIII (FVIII).
                     rather than CD16+CD14+ monocytes were prone to dif-            Current HA therapies, albeit significantly improving HA




                                                                                                     on
                     ferentiate into osteoclasts. The purpose of this study is to   treatment, are mainly aimed at replacing the missing fac-
                     better understand how the osteoclastogenesis could be          tor and do not provide a definitive cure. On the other
                     influenced by the FVIII, von Willebrand Factor (VWF)           hand, gene therapy, albeit providing promising results,




                                                                                               e
                     and thrombin and to assess the osteoclastogenic poten-         still have significant drawbacks and is far from being
                     tial from two different HA patients. Methods:
                     Osteoclastogenesis of healthy donors-derived PBMC
                     (Peripheral Blood Mononuclear Cells) were assessed in
                                                                                        us
                                                                                    translated into the clinic. Genome editing approaches
                                                                                    based on homologous recombination (HSR) have been
                                                                                    also explored but are not optimized yet, with still safety
                                                                                    concerns to be addressed. Recently, new genome editing
                                                                                    al
                     the presence of plasma derived VWF/FVIII complex,
                     rVWF, rFVIII and thrombin. Moreover, in order to study         approaches non-relying on HSR, namely Base Editing
                                                                            ci

                     cell alteration in HA patients, osteoclastogenesis was         (BE) and Prime Editing (PE) strategies, have been devel-
                     performed with PBMC isolated from two HA patients              oped and showed their ability to efficiently and cleanly
                                                                      er


                     and compared with healthy controls. Moreover, FACS             install or revert point mutations which, in the HA field,
                     analysis was performed in order to study circulating           are the most represented ones when excluded the IVS22
                                                                m



                     osteoclast precursors, of adult haemophilic patient.           inversion. The BE and PE approaches, based on engi-
                     Results: The treatment of PBMC with coagulation fac-           neered Cas proteins fused with nucleotide deaminase or
                                                   om




                     tors reduced osteoclastogenesis even at levels compara-        reverse transcriptase domains, respectively, are able to
                     ble to that induced by osteoprotegerin. In particular,         efficiently and selectively modify the target nucleotide
                     VWF showed a key role in regulation of osteoclasts dif-        without inducing any double strand breaks or recombina-
                                                -c




                     ferentiation process, inhibiting ~43% of the osteoclasto-      tion events. The aim of this study is to develop BE and
                     genesis while, when VWF was complexed with FVIII,              PE tools to revert a panel of frequent F8 missense and
                                        on




                     the 50% of inhibition was observed. On the other hand,         nonsense variants associated with severe HA phenotypes
                     thrombin inhibits osteoclast differentiation (up to 95%        and thus rescue FVIII expression. Methods: Design and
                     of inhibition). PBMC isolated from both severe/pedi-           cloning of BE/PE components as well as of FVIII cas-
                                   N




                     atric and mild/adult haemophilic patients, showed an           settes to create expression vectors; Creation of cellular
                     increased ability to produce mature osteoclasts in com-        models by transient and stable expression of recombinant
                     parison to those obtained from healthy donor. Osteoclast       FVIII variants; Delivery of genome editing actors by
                     precursors (CD16−CD14+CD11b+) are significantly                lipofection and evaluation of rescue at DNA (DNA
                     higher in adult/mild HA patient than age and sex               sequencing) and protein (antigen by ELISA and activity
                     matched controls (~33%), whereas the percentage of             by aPTT assay) levels. Results: c.6046 C>T
                     CD16+ cells was reduced. According to these data, gene         (p.R2016W), c.6496 C>T (p.R2166*), c.6545 G>A
                     expression analysis on RNA extracted from patient’s and        (p.R2182H), c.6682 C>T (p.R2228*), c.6683 G>A
                     control’s derived mature osteoclasts, revealed high-lev-       (p.R2228Q), through transient and stable expression
                     els of mRNA levels of RANK, TRAF6, CATHEPSIN-K                 studies, are associated with reduced secreted FVIII levels
                     and TCIRG1 compared to matched controls.                       (p.R2166* 0%; p.R2228Q 13%; p.R2016W 39%;
                     Conclusions: All these data support that bone loss             p.R2182H 65% of WT counterpart). On the other hand,
                     observed in haemophilic patients could be related to           the p.R2228* change resulted in appreciable FVIII pro-
                     increased osteoclast formation and activity and that           tein levels (84% of WT ) likely attributable to the secre-
                     coagulation factors directly impact on bone cells.             tion of a truncated FVIII isoform. Studies on FVIII activ-
                     Further studies are needed in order to translate the           ity revealed that p.R2228Q maintains the same specific
                     results into benefits for clinical practice of haemophilia     activity of the wt counterpart, indicating in the secretion
                     patients suffering from bone disease.                          defect the major cause of patient phenotype. We identi-




         [page 46]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 47



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    fied, through a screening process, a panel of BE/PE able     definition by co-transfection of mutant minigene with
                    to rescued FVIII secretion for the p.R2166*, p.R2182H        compensatory (n=1) or Exon Specific U1snRNA (n=3)
                    and p.R2228Q variants. Additional studies on stable          followed by splicing pattern analysis with plasmid-spe-
                    clones expressing the p.R2166* and p.R2228Q muta-            cific amplicons. Results: Bioinformatic analysis did not
                    tions demonstrated that the BE and PE tools were able to     predict aberrant splicing due to the F8 c.1752+5G>C
                    revert the mutations at DNA levels and resulted in signif-   mutation. However, splicing assays in different
                    icant rescue (up to 20% of FVIII-WT) of secreted FVIII       hepatoma cells demonstrated that exon 11 is well
                    protein and activity levels. Conclusions: Overall, for the   defined, as confirmed by the complete exon inclusion in
                    first time we applied BE and PE to frequent FVIII point      the pIVS11wt context, and that the c.1752+5G>C change
                    mutations leading to severe Haemophilia A.                   induces exon 11 skipping, to an extent that depends on
                    Experimental data provided the proof-of-principle of         the transfected cell lines. In the worst scenario, the muta-
                    efficacy in cellular models, which are currently under       tion is associated with low levels of correctly spliced
                    validation in HA blood-outgrowth endothelial cells           transcripts (~10%). Notably, co-transfection of different
                    (BOECs) and planned in mouse models.                         engineered U1snRNA significantly improves FVIII exon
                                                                                 11 definitions and thus inclusion, with the compensatory
                                                                                 U1snRNA associated with exon 11 inclusion up to 92%.
                    OC067                                                        Conclusions: Overall, we provide experimental evi-
                                                                                 dence that the F8 c.1752+5G>C change leads to exon
                                                                                 skipping by impairing proper exon 11 definition and is




                                                                                                          ly
                    RESCUE OF A HEMOPHILIA A-CAUSING FVIII
                                                                                 associated with trace levels of correctly spliced tran-
                    SPLICING VARIANT VIA ENGINEERED




                                                                                                  on
                                                                                 scripts, in accordance with the HA patients’ phenotype.
                    U1SNRNAS                                                     Moreover, the splicing outcome is cell-dependent, and
                                                                                 further studies aimed at identifying the involved splicing
                    L. Peretto1, R. Tarantino1, A. Borchiellini2,                factor will be conducted. Notably, F8 exon 11 inclusion




                                                                                            e
                    F. Bernardi1, A. Follenzi3, M. Pinotti1, D. Balestra1        can be efficiently restored by RNA therapeutics based on
                    1Department    of Life Sciences and Biotechnology,
                    University of Ferrara, Ferrara, Italy; 2Citta della salute
                                                                                     us
                                                                                 engineered U1snRNAs, which are currently under inves-
                                                                                 tigation through lentiviral-mediated delivery in Blood
                                                                                 Outgrowth Endothelial Cells (BOEC) isolated from HA
                    e della scienza, Torino, Italy; 3Department of Health
                                                                                 al
                                                                                 patients.
                    Sciences, University of Piemonte Orientale, Novara, Italy
                                                                            ci

                    Background and Aims: Hemophilia A (HA), the most
                                                                                 OC068
                                                                   er


                    common coagulation bleeding disorder (1:5000 males),
                    is an X-linked hereditary disease caused by F8 gene
                                                             m



                    mutations leading to deficiency of factor VIII (FVIII).      INFLUENCE OF EMICIZUMAB OF PROTEIN
                    Current HA therapies, albeit significantly improving HA      C-MEDIATED CLOTTING REGULATION
                                                 om




                    treatment, are mainly aimed at replacing the missing fac-
                    tor and do not provide a definitive cure. Among all HA-      F. Mancazzo, L. Dirienzo, A. Vitulli, C.T. Ammollo,
                    causing mutations, those affecting splicing are relatively   F. Semeraro, M. Colucci
                    frequent, particularly in the severest forms. These muta-
                                              -c




                    tions are generally associated with exon skipping and can    Dipartimento di Scienze Biomediche e Oncologia
                                                                                 Umana, Università di Bari “Aldo Moro”, Italy
                                       on




                    be potentially rescued by RNA therapeutics. Among
                    these, variants of the key U1snRNA spliceosomal com-
                    ponent have been shown to efficiently rescue exon inclu-     Background and Aims: Emicizumab is a bispecific anti-
                                  N




                    sion impaired by mutations located at the 3’splice site      body which functions as FVIII-mimetic by simultane-
                    (ss), 5’ss or within the defective exon, in both cellular    ously binding FIXa and FX. At variance with plasma
                    and animal models of human disease. This study aims to       FVIII, emicizumab is insensitive to degradation by acti-
                    dissect the molecular mechanism underlying the moder-        vated protein C (APC) and may thus tilt the coagulation
                    ate HA (FVIII:C<5%) phenotype identified in two broth-       balance towards a prothrombotic state. We investigated
                    ers carrying the F8 c.1752+5G>C variant and to develop       the effect of emicizumab on PC-mediated inhibition of
                    RNA therapeutics based on engineered U1snRNAs to             coagulation under in vitro conditions mimicking physio-
                    restore proper exon 11 definition. Methods: Creation of      logical and pathological clotting activation. Methods:
                    expression vectors for the wild-type (pIVS11wt) and          Hemophilic plasma (<1% FVIII) was supplemented with
                    mutant (pIVS11+5G>C) F8 minigenes and for the engi-          emicizumab (50 or 100 µg/ml) or recombinant FVIII
                    neered U1 snRNAs designed to base pairs to the mutated       (Kovaltry, 1 IU/ml). Thrombin generation was assessed
                    5’ss (compensatory U1snRNA) or to less-conserved             by CAT assay using as clotting trigger tissue factor (TF,
                    downstream intronic sequences (Exon Specific                 1 pM) or an intrinsic pathway activator (1/50 diluted
                    U1snRNA). Transient expression of F8 minigenes, either       aPTT reagent or 20 pM FXIa). The effect of the PC sys-
                    wild-type (pIVS11wt) or harbouring the variant               tem was assessed by adding APC (0.2-0.8 µg/ml) or by
                    c.1752+5G>C (pIVS11+5G>C), in different hepatoma             activating plasma PC by thrombomodulin (TM, 4 nM) or
                    cell lines (Huh7, HepG2, Hepa1-6) as well as HEK293T,        endothelial cells (EA.hy926, 50,000/well). In some
                    followed by splicing pattern analysis. Rescue of exon 11     experiments, FXa generation was assessed by a chro-




                                                              [Hematology Reports 2021; 13:s3]                                                  [page 47]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 48



         Oral communications




                     mogenic substrate (S-2732). Results: In TF-triggered         protective levels for bleeding, while avoiding excessive
                     coagulation (physiologic activation), emicizumab-plas-       levels with inherent thrombotic risk. The recent availabil-
                     ma displayed a good response to added APC as well as to      ity of the bi-specific monoclonal antibody emicizumab
                     endogenous PC activation by TM or endothelial cells          for long-term prophylaxis in PWSHAhas opened a new
                     addition, with a clear-cut inhibition of thrombin genera-    scenario in planning antihemorrhagic perioperative treat-
                     tion, which was comparable to that observed in FVIII-        ment with FVIII with its potential risks. The aim of the
                     plasma. When coagulation was triggered through the           present study is to report our experience in management
                     intrinsic pathway (pathologic activation), the prolonga-     of surgical procedures in PWSHA on emicizumab.
                     tion of lag-time by increasing concentrations of APC was     Methods: Between January 2018 and April 2022, 17
                     less pronounced in emicizumab-plasma than in FVIII-          PWHA underwent 34 surgical procedures (5 major sur-
                     plasma, whereas the reduction of thrombin peak and ETP       geries, 29 minor surgeries).All major surgeries were
                     was similar. Using the latter condition, we found that the   managed under FVIIIcoverage, with factor activity levels
                     generation of FXa, which persisted after prothrombin         of at least 50 IU/dL maintained for 7-14 days. Adjunctive
                     consumption, was significantly greater in emicizumab-        intravenous tranexamic acid (TA) 1 g every 12 hours for
                     plasma and, contrary to FVIII-plasma, barely affected by     7 days was also administered. Thromboprophylaxis was
                     APC (Figure 1). Finally, when purified prothrombin was       not prescribed. Minor surgeries were managed with a
                     added to (defibrinated) plasma after consumption of          rFVIII preoperative bolus (25-50 IU/kg), except for tooth
                     endogenous prothrombin, the second wave of thrombin          extractions, dental procedures, and cataract, performed
                     generation in emicizumab-plasma was greater and              without rFVIII coverage. In dental surgeries TA has been




                                                                                                          ly
                     markedly less affected by APC as compared to FVIII-          used as mouthwash. When clinically appropriate, FVIII
                                                                                  activity levels were monitored by using a bovine chro-




                                                                                                   on
                     plasma, likely because of the higher concentration of
                     FXa in the former. Conclusions: In vitro, in a “closed”      mogenic assay. Results: No thromboembolic events have
                     system, thrombin generation in emicizumab-plasma was         occurred. No transfusional support was needed. Bleeding
                     effectively inhibited by the PC system (with the only        was as expected depending on the type of surgery (Table




                                                                                             e
                     exception of a weaker lag-time prolongation in contact-      1). Conclusions: Major surgery with a regimen of emi-
                     phase activated plasma), suggesting a major role of FVa
                     inactivation by APC. However, the greater generation of
                     FXa in emicizumab-plasma in the presence of APC
                                                                                      us
                                                                                  cizumab and rFVIII in PWSHA has been safely and effi-
                                                                                  caciously performed. Minor surgeries can be safely per-
                                                                                  formed without prophylaxis with rFVIII. A multidiscipli-
                                                                                  nary team and strict monitoring are recommended for the
                                                                                  al
                     might translate in vivo (“open” system) in a greater and
                     sustained thrombin generation thanks to the continuous       management of surgical procedures in PWSHA treated
                                                                          ci

                                                                                  with emicizumab.
                     supply of prothrombin.
                                                                    er


                                                                                  Table 1.
                                                               m
                                                   om
                                                -c
                                        on




                     Figure 1.
                                   N




                     OC069

                     SURGICAL PROCEDURES IN SUBJECTS WITH
                     SEVERE HEMOPHILIA A TREATED WITH
                     EMICIZUMAB: A SINGLE CENTRE EXPERIENCE

                     S. Linari, L. Pieri, F. Demartis, R. Fjerza,
                     M.C. Di Stefano, G. Castaman

                     Centre for Bleeding Disorders and Coagulation,
                     Department of Oncology, Azienda Ospedaliero-
                     Universitaria Careggi, Florence, Italy

                     Background and Aims: The management of surgery in
                     persons with severe haemophilia A (PWSHA) requires
                     intensive treatment with factor VIII (FVIII) and postop-
                     erative close monitoring of circulating FVIII to maintain




         [page 48]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 49



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    OC070                                                       autoantibodies significantly reduced the percentage of
                                                                                PPF on fibrinogen (27.7±2.8%) compared to control
                                                                                serum (41.9±4.2%; p<0.05) while no differences were
                    ANTI-INTEGRIN GPIIBIIIA IMMUNE
                                                                                found on collagen type I. Conclusions: The role of
                    THROMBOCYTOPENIA AUTOANTIBODIES IMPAIR                      αIIbβ3 in MK spreading is well known, indeed αIIbβ3
                    MEGAKARYOCYTE MIGRATION, POLARIZATION                       antagonists block spreading on fibrinogen. The role of
                    AND PROPLATELET FORMATION                                   αIIbβ3 in PPF is instead debated (Balduini et al. J
                                                                                Thromb Haemost. 2008;6:1900-7; Bury et al. PLoS
                    E. Giglio, E. Petito, L. Bury, A. Mezzasoma, P. Gresele     One. 2012;7:e34449). We show that anti-αIIbβ3 autoan-
                                                                                tibodies not only impair PPF formation by MKs but
                    Section of Internal and Cardiovascular Medicine,            also prevent MK polarization and migration, two cru-
                    Department of Medicine, University of Perugia, Perugia,     cial steps for the release of platelets in the blood stream.
                    Italy                                                       Our results therefore unravel a new pathogenic mecha-
                                                                                nism of anti-αIIbβ3 autoantibodies possibly explaining
                    Background and Aims: Immune thrombocytopenia                thrombocytopenia that characterizes a subgroup of ITP
                    (ITP) is an autoimmune disorder characterized by iso-       patients, suggesting that ectopic platelet release, besides
                    lated thrombocytopenia due to antiplatelet autoantibod-     impaired PPF, may participate in thrombocytopenia
                    ies against platelet surface glycoproteins (αIIbβ3-         induced by anti-αIIbβ3 autoantibodies.
                    GPIIb/IIIa, α2β1-GPIa/IIa, GPIb/IX/V) leading to




                                                                                                         ly
                    increased platelet clearance. Recent data however show
                    that they bind surface glycoproteins also on megakary-




                                                                                                 on
                                                                                OC071
                    ocytes (MKs) possibly impairing platelet production.
                    Both MK migration from the osteoblastic to the vascu-
                    lar niche and MK polarization are crucial steps in          THE ITALIAN REGISTRY ON ACTIVE ADULT ITP:




                                                                                            e
                    thrombopoiesis and their defect, with consequent            UPDATE AND PRELIMINARY RESULTS ON
                    ectopic platelet production in the bone marrow, has
                    been reported to contribute to thrombocytopenia in
                    some congenital thrombocytopenias. Defective MK
                                                                                    us
                                                                                QUALITY OF LIFE

                                                                                L. Ghiotto1, N. Vianelli2, G. Carli3, G. Auteri2,
                                                                                M. Carpenedo4, A. Ricco5, E. Rossi6, V. De Stefano6,
                                                                                al
                    adhesion, spreading and proplatelet formation (PPF)
                    induced by anti-αIIbβ3 and anti-αIIbβ3+anti-α2β1            V. Carrai7, F. Morelli7, W. Barcellini8, B. Fattizzo8,
                                                                                A. Chistolini9, C. Santoro9, S. Cantoni10,
                                                                        ci

                    autoantibodies were previously reported (Grodzielski et
                    al. Br J Haematol. 2018;183:319-323) but no studies on      A. Borchiellini11, F. Valeri11, U. Consoli12, V. Innao12,
                                                                  er


                    MK migration and polarization were carried out. Our         G. Gaidano13, A. Patriarca13, E. Rivolti14, V. Pavone15,
                    aim was to evaluate the effects of anti-αIIbβ3              S. Sibilla15, F. Pane16, F. Chiurazzi16, S. Siragusa17,
                                                             m



                    antiplatelet autoantibodies detected in serum of a series   M. Napolitano17, F. Zaja18, E. Lucchini18, A. Poloni19,
                    of ITP patients on MKs, focusing on their ability to        D. Lame19, C. Bosi20, D. Lazzarotto21, O. Mulas22,
                                                 om




                    modulate some crucial steps of thrombocytopoiesis,          L. Pontalto1, E. Crea23, P. Fazi23, F. Rodeghiero1
                    like MK migration, polarization, spreading and PPF.
                                                                                1Fondazione    Progetto Ematologia, Vicenza, Italy;
                    Methods: Anti-αIIbβ3 autoantibodies were detected by
                                                                                2Istitutodi Ematologia “L. e A. Seragnoli” - Policlinico
                                              -c




                    the MAIPA assay in serum from 10 ITP patients.
                    Control serum was obtained from 10 healthy subjects.        “S. Orsola Malpighi”, Università degli Studi di Bologna,
                                                                                Bologna, Italy; 3Divisione Ematologia, Ospedale “S.
                                      on




                    Human MKs were differentiated from peripheral blood-
                    derived CD34+ cells from healthy subjects. MK migra-        Bortolo”, Vicenza, Italy; 4Unità di Ematologia con
                    tion towards SDF-1α was assessed by a transwell sys-        Trapianto, ASST “S. Gerardo”, Monza, Italy; 5UO
                                  N




                    tem, MK polarization on fibrinogen by fluorescence          Ematologia con Trapianto, Policlinico Universitario
                    microscopy upon staining with anti-CD42b MoAb and           “Giovanni XXIII”, Università degli Studi di Bari “Aldo
                    Hoechst for nuclei, MK spreading and PPF on fibrino-        Moro”, Bari, Italy; 6Istituto di Ematologia, Università
                    gen and on type I collagen by fluorescence microscopy       Cattolica del Sacro Cuore, Policlinico “A. Gemelli”,
                    upon staining with anti-β1-tubulin antibody for micro-      Roma, Italy; 7SOD Ematologia, AOU Careggi, Firenze,
                    tubules, rhodamine-phalloidin for actin and Hoechst for     Italy; 8UOC Oncoematologia, Fondazione IRCCS Ca’
                    nuclei. All data are reported as mean±SEM. Results:         Granda Ospedale Maggiore Policlinico, Milano, Italy;
                                                                                9Divisione di Ematologia, Policlinico Universitario
                    The area of the bottom face of the transwell filter cov-
                    ered by MKs was significantly reduced (32%; p<0.01)         “Umberto I”, Università degli Studi “Sapienza”, Dip.
                    upon coincubation with anti-αIIbβ3 ITP serum com-           Biotecnologie Cellulari ed Ematologia, Roma, Italy;
                                                                                10Dipartimento di Ematologia e Oncologia, Niguarda
                    pared to control serum showing impaired MK migra-
                    tion. The percentage of polarized MKs on fibrinogen         Cancer Center, Azienda Socio Sanitaria Territoriale
                    was significantly reduced upon coincubation with anti-      Ospedale Niguarda, Milan, Italy; 11Divisione di
                    αIIbβ3 ITP serum compared with control serum                Ematologia dell’Università degli Studi di Torino, “Città
                    (15.6±1.3% vs 36.2±2%; p<0.01). The percentage of           della Salute e della Scienza di Torino”, Torino, Italy;
                                                                                12UOC Ematologia, PO Nesima, ARNAS “Garibaldi”,
                    spread MKs in the presence of anti-αIIbβ3 ITP serum
                    compared to control serum was not impaired, either on       Catania, Italy; 13Ematologia, AOU Maggiore della
                    fibrinogen or on type I collagen. However, anti-αIIbβ3      Carità, Dipartimento di Scienze Mediche, Università del




                                                             [Hematology Reports 2021; 13:s3]                                                  [page 49]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 50



         Oral communications




                     Piemonte Orientale “Amedeo Avogadro”, Novara, Italy;           2 menorrhagia, 1 severe menorrhagia, 2 gastrointestinal
                     14Dipartimento di Ematologia, Azienda Unità Sanitaria          bleeding, 1 intracranial bleeding. Thromboembolic
                     Locale (AUSL), IRCCS Reggio Emilia, Italy; 15UOC               events occurred in 47 patients (8%). By a preliminary
                     Ematologia e TMO, AO “Card. G. Panico”, Tricase                analysis, QoL and Fatigue scores appear lower than in
                     (LE), Italy; 16Dipartimento di Ematologia e Trapianto del      normals. In multivariate analysis considering patients
                     Midollo Osseo, Azienda Ospedaliera Universitaria               without missing data, the absence of general risk factors
                     “Federico II”, Napoli, Italy; 17UO di Ematologia con           for thrombosis was significantly associated with fatigue
                     Trapianto, Dipartimento Biomedico di Medicina Interna,         and both SF-12 scores. Sport practice was significantly
                     Azienda Ospedaliera Universitaria “Paolo Giaccone”,            associated with fatigue and MCS-12. Female gender
                     Palermo, Italy; 18Ematologia, Azienda Sanitaria                was significantly associated with fatigue and PCS-12.
                     Universitaria Integrata, Trieste, Italy; 19Clinica di          No statistically significant association was found
                     Ematologia, Università Politecnica delle Marche e AOU          between HRQoL measures and patient condition at
                     Ospedali Riuniti di Ancona Italy; 20Dipartimento di            entry. SMOG index and splenectomy status also did not
                     Oncologia-Ematologia, Azienda USL Ospedale di                  influence HRQoL and fatigue. Absence of association
                     Piacenza, Italy; 21Ematologia e Trapianto di Cellule           between platelet count at enrollment and both fatigue
                     Staminali, Azienda Sanitaria Universitaria Friuli              and HRQoL measures was also observed. Conclusions:
                     Centrale (ASUFC), Udine, Italy; 22SC Ematologia e              Our data confirm that patients seen in current clinical
                     CTMO, Ospedale Businco, ARNAS “G. Brotzu”,                     practice have an increased incidence of thromboembol-
                     Cagliari, Italy; 23Fondazione GIMEMA Onlus, Roma,              ic events and that most patients show minor bleeding




                                                                                                              ly
                     Italy                                                          symptoms with a moderate negative impact on QoL.
                                                                                    The enrollment seems to have slowed down with the




                                                                                                      on
                     Background and Aims: Registries are essential for              advent of the COVID-19 pandemic with the number of
                     clinical and patient-report data in rare diseases. In          new diagnoses probably underestimated. A linkage with
                     Europe, 3 population-based and 12 clinical cohorts of          the existing European ITP registries is planned.




                                                                                                e
                     ITP patients, including the Italian ITP registry, have
                     been identified by the European Research Consortium
                     on ITP (www.ercigroup.org) (Moulis et al. BJHaem
                     2021). The Italian registry, sponsored by GIMEMA
                                                                                        us
                                                                                    Table 1. Main characteristics of the 576 evaluable patients.
                                                                                    al
                     Foundation (Rome) and managed by the Hematology
                     Project Foundation (Vicenza), started in late 2018 and
                                                                            ci

                     is aimed at producing a dynamic picture of adult ITP
                     natural history as modified by management by collect-
                                                                     er


                     ing and analyzing standardized data. Variables, includ-
                     ing comorbidities, safety and efficacy of treatments and
                                                                m



                     QoL will be considered. Methods: Multicenter obser-
                     vational (prospective and retrospective) study, includ-
                                                   om




                     ing a 3-year enrollment period with a subsequent follow
                     up of at least 2 years, recruiting patients on active treat-
                     ment. Patient conditions at entry were classified as: ini-
                                                -c




                     tiating a first-line (group A); requiring modification of
                     previous treatment (group B); being on treatment
                                        on




                     (group C). Historical data are collected at entry, while
                     prospective data at annual visits. Data quality is moni-
                     tored and assured by electronic queries and interactions
                                   N




                     between a central team and recruiting centers. To pro-
                     duce a dynamic individual picture, each patient is
                     tracked as it moves through different disease statuses
                     (complete response, response, no response, toxicity, not
                     on treatment) according to the various treatment lines.
                     Results: As to May 10, 2022, 676 patients were sequen-
                     tially recruited (195 more than the previous report of
                     May 2020) by 21 active centers. 576 cases were fully
                     evaluable regarding the variables of interest. Main
                     patients’ characteristics and data on bleeding, thrombo-
                     sis and QoL at entry are reported in the Table 1. Among
                     patients with at least a grade 1 bleeding using the Skin
                     Mucosal Organ Grading system (SMOG) (Rodeghiero
                     et al. Blood 2013) at entry or in the previous month, 47
                     patients had only skin, 5 had only mucosal, 16 had both
                     skin and mucosal, 2 had both skin and organ and 4 had
                     only organ bleeding. Most S≥2 (N=16) were petechiae.
                     Most M≥1 (N=21) were in oral cavity. O≥1 (N=6) were




         [page 50]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 51



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    OC072                                                        low titer anti-ADAMTS13 IgGs (17 U/ml) and treated
                                                                                 with plasma-exchange every other day for nine months,
                                                                                 rituximab, vincristine, cyclophosphamide and splenec-
                    ANTI-ADAMTS13 ANTIBODIES IN CONGENITAL                       tomy. Conclusions: Neutralizing anti-ADAMTS13 IgG
                    TTP PATIENTS                                                 could develop very rarely in cTTP patients after pro-
                                                                                 longed exposure to plasma, exogenous source of
                    I. Mancini1, M. Biganzoli2, A. Cairo2, M. Capecchi2,3,       ADAMTS13. Monitoring of anti-ADAMTS13 antibod-
                    A. Artoni2, B. Ferrari2, S.M. Trisolini4, L. Facchini5,      ies in cTTP patients with a reduced therapeutic response
                    G. Giuffrida6, A. De Fanti7, P. Agosti1, F. Peyvandi1,2      is recommended.
                    1Università   degli Studi di Milano, Department of
                    Pathophysiology and Transplantation, and Fondazione          OC073
                    Luigi Villa, Milan, Italy; 2Fondazione IRCCS Ca’ Granda
                    Ospedale Maggiore Policlinico, Angelo Bianchi Bonomi
                    Hemophilia and Thrombosis Center, Milan, Italy;              RISK FACTORS FOR SARS-COV-2 INFECTION AND
                    3Università degli Studi di Milano, Department of             IMPACT OF DISEASE IN A MONOCENTRIC
                    Biomedical Sciences for Health, Milan, Italy;                COHORT OF PATIENTS WITH ACTIVE IMMUNE
                    4Hematology, Department of Translational and Precision
                                                                                 THROMBOCYTOPENIA (ITP)
                    Medicine, “Sapienza” University of Rome, Rome, Italy;




                                                                                                         ly
                    5Hematology Unit, Azienda Unità Sanitaria Locale,
                                                                                 G. Carli, G. Marchesini, I. Nichele, A. Tosetto
                    IRCCS di Reggio Emilia, Reggio Emilia, Italy;




                                                                                                 on
                    6Hematology Division, Department of Clinical and
                                                                                 Hemophilia and Thrombosis Center, Hematology
                    Molecular Biomedicine, University of Catania, Catania,       Department, SBortolo Hospital, Vicenza, Italy
                    Italy; 7Departmental Simple Unit of Pediatric




                                                                                            e
                    Rheumatology, AUSL-IRCSS Reggio Emilia, Reggio               Background: After the outbreak of COVID-19, several
                    Emilia, Italy

                    Background: Congenital thrombotic thrombocy-
                                                                                     us
                                                                                 cases of de novo ITP secondary to SARS-CoV2 infec-
                                                                                 tion or COVID-19 vaccination have been reported.
                                                                                 Little is known about the impact of SARS-CoV-2 infec-
                    topenic purpura (cTTP) is an autosomal recessive dis-        tion in patients with active ITP. Aim: We aimed to
                                                                                 al
                    ease due to mutations in the ADAMTS13 gene. cTTP             assess risk factors for SARS-CoV-2 infection and the
                                                                         ci

                    manifests as acute episodes of thrombotic microan-           effect of COVID-19 infection on ITP clinical course
                    giopathy characterized by ADAMTS13 activity <10              treatment in a monocentric cohort of ITP patients with
                                                                   er


                    IU/dl and absence of anti-ADAMTS13 antibodies,               active disease. Materials and Methods: We included
                    which are treated by plasma infusion to replenish the        all patients with active ITP (either newly diagnosed,
                                                             m



                    deficient protease. cTTP patients with anti-                 persistent, or chronic ITP, with ongoing therapy and
                    ADAMTS13 antibodies were rarely described. Aims:             platelet counts <100 x109/L) on regular follow-up from
                                                 om




                    To investigate anti-ADAMTS13 IgG in our cTTP                 February 2020 to May 2022. Either antigenic or molec-
                    cohort. Methods: We performed a cross-sectional study        ular nasal swab confirmed SARS-CoV-2 infection. We
                    of cTTP patients with homozygous or compound-het-            retrieved data on a previous splenectomy, exposure to
                    erozygous ADAMTS13 mutations, enrolled in the
                                              -c




                                                                                 rituximab up to one year before the pandemic, or ongo-
                    Milan TTP registry from 2002 to 2018. Plasma samples         ing steroid or immunosuppressant therapy. In pts with
                                       on




                    collected during the acute or remission phase were test-     SARS-CoV-2 infection, we recorded the severity of
                    ed for anti-ADAMTS13 IgG using an in-house western           infection (WHO classification), need for hospitaliza-
                    blot or ELISA based assay or the Technozym                   tion, specific treatment for COVID-19, need to modify
                                  N




                    ADAMTS13 Inhibitor ELISA kit (Technoclone).                  ITP treatment, bleeding, and thrombotic events.
                    Neutralizing activity of anti-ADAMTS13 antibodies            Results: Between Feb 2020 and May 2022, 88 pts were
                    was measured using a FRET-based Bethesda-like assay.         regularly followed up in our center for active ITP. 29 pts
                    Results: Ninety-one samples from 27 cTTP patients            (32.9%) developed SARS-CoV-2 infection (Table 1).
                    (52% females, median age at onset 6.5 years [interquar-      Patients with a history of exposure to rituximab were
                    tile range 1.2–20.1]) were tested for anti-ADAMTS13          more frequently infected in our cohort (p=0.002).
                    IgG (samples per patient, median [range]: 2 [1-13]).         Patients with SARS-CoV-2 infection showed a mild
                    Four acute phase samples from four cTTP patients were        disease in 90% of cases (26 pts), while three patients
                    positive by at least two anti ADAMTS13 IgG assays. In        (10%) needed to be hospitalized for severe pneumonia
                    one case, non-neutralizing anti-ADAMTS13 IgG (83             (one with ARDS and need for intubation). Two hospi-
                    U/ml) developed during ticlopidine treatment, consid-        talized patients were treated with hyperimmune plasma
                    ered the trigger of the acute event. Two patients present-   because of recent exposure to rituximab. One of them
                    ed non-neutralizing anti-ADAMTS13 IgG (33 and 19             experienced after one year a SARS-CoV-2 reinfection
                    U/ml) after infusion of four units of fresh frozen plasma    requiring treatment with monoclonal antibodies as an
                    over 33 and 59 days, respectively. The last case present-    outpatient. No patient died from COVID-19. Three
                    ed extremely high titers of anti-ADAMTS13 IgG (8800          patients at home have been treated with antiretroviral
                    U/ml) with neutralizing activity (295 BU/ml) after           therapy (nirmatrelvir/ritonavir) and two patients with
                    being misdiagnosed with acquired TTP due to baseline         monoclonal antibodies (bamlavinimab/etesevimab)




                                                              [Hematology Reports 2021; 13:s3]                                                [page 51]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 52



         Oral communications




                     with no side effects and rapid resolution of symptoms.     chronic ITP after COVID-19 infection and vaccination.
                     No patients experienced significant bleeding events,       Methods: All pts currently followed for chronic ITP
                     and the great majority (27 patients, 93%) maintained       who experienced COVD19 infection and/or who com-
                     the same therapy for ITP (TPOra in 40% of cases). One      pleted first and second dose vaccination with an avail-
                     hospitalized patient needed to acutely start treatment     able plt count before and after such events were includ-
                     (steroid plus immunoglobulins) for a significant drop in   ed in the analysis. Demographic features, ITP duration,
                     platelet count with subsequent partial response. Another   number and type of previous and current treatment;
                     outpatient has stopped therapy with rituximab (after       reduction in plt count of any grade, described as
                     three doses) and has been treated with antiretroviral      absolute and percentage reduction were recorded.
                     therapy, maintaining a complete response on the            Finally, ITP exacerbations, defined >50% decline in plt
                     platelet count. No thrombotic adverse events have been     count compared with baseline, >20% decline in plt
                     recorded. Conclusions: Previous exposure to rituximab      count compared with baseline and a plt nadir
                     appears to be an important risk factor for SARS-CoV-2      <30.000/ul or need for rescue medication) were also
                     infection in patients with active ITP. In most cases,      included. Proportion comparisons were carried out with
                     COVID-19 had a benign course without consequences          Chi square test. Results: 64 chronic ITP cases were
                     on the ITP course, but patients with an immunodeficient    included in the analysis. Study population characteris-
                     status (e.g., after Rituximab exposure) can be prone to    tics are resumed in Table 1.
                     more severe disease.
                                                                                Table 1.




                                                                                                        ly
                     Table 1.




                                                                                                 on
                                                                                           e
                                                                                    us
                                                                                al
                                                                         ci
                                                                    er



                                                                                Before any vaccination, 11 pts got COVID19 infection.
                                                              m




                     OC074                                                      A reduction in plt count was observed in 6 patients
                                                                                (54,5%) with median plt count after infection of
                                                  om




                                                                                4.000/ul (IQR1 2000- IQR3 46500/ul), mean absolute
                     RISK OF ITP EXACERBATION IN CHRONIC ITP                    plt reduction of 122.166/ul (IQR1 58.000 – IQR3
                     PATIENTS AFTER COVID19 INFECTION AND                       157.750/ul) and mean percentage reduction of 92%
                                               -c




                     VACCINATION: A SINGLE-CENTER                               (IQR1 67.37 - 97.50%). ITP exacerbation was observed
                     OBSERVATIONAL STUDY                                        in 5 pts (45,5%), all cases requiring rescue therapy. First
                                        on




                                                                                vaccine dose was received by 61 pts; an available plt
                     C. Dainese1, F. Valeri1, M. Bardetta1, C. Sella1,          count after first dose was available for 41 pts after a
                     B. Bruno2, A. Borchiellini1                                mean time of 11,4 days (2 – 15 days). A reduction in plt
                                   N




                                                                                count was observed in 17 pts (41.4%) with a mean plt
                     1Centro Riferimento Regionale per Malattie trombotiche
                                                                                count after first dose of 147.736/ul (IQR1 72250 - IQR3
                     ed Emorragiche dell’adulto, Ematologia Universitaria,      220750/ul), a mean absolute reduction of 53058/ul
                     AOU Città della Salute e della Scienza di Torino;          (IQR1 9000 - IQR3 70500/ul) and a mean percentage
                     2Ematologia Universitaria, Dipartimento di Oncologia,
                                                                                reduction of 30% (IQR1 5.2 - IQR3 49.75%). An ITP
                     AOU Città della Salute e della Scienza di Torino, Italy    exacerbation after first vaccine dose was observed in 6
                                                                                pts (14.63%) with 3 pts (7.31%) requiring rescue treat-
                     Background and Aims: Current Sars-Cov 2 disease            ment. 58 pts received second vaccine dose; an available
                     (COVID 19) pandemic is characterized by severe pneu-       plt count after second dose was available for 36 pts after
                     monia, multi-organ disease and dysregulation of immu-      a mean time of 16.8 days (8 –21 days). A reduction in
                     nity. Vaccination is one of the most effective weapons     plt count was observed in 16 pts (44.4%) with a mean
                     to control the disease. Both SARS-CoV-2 infection and      plts count after first dose of 147.615,38/ul (IQR1
                     vaccination have raised concern in immune mediated         77.000/ul – IQR3 201.750/ul), a mean absolute reduc-
                     diseases, including immune thrombocytopenic purpura        tion of 77.437/ul (IQR1 14.500 - IQR3 66.750/ul) and a
                     (ITP) considering the risk of de novo ITP development      mean percentage reduction of 28% (IQR1 6.75 -
                     and ITP recurrence. Here, we report data from a single     40.25%). An ITP exacerbation after second vaccine
                     Center retrospective-prospective collection aiming to      dose was observed in 5 pts (13.88%), with 2 pts (5,5%)
                     evaluate platelet (plt) dynamics in patients (pts) with    requiring rescue treatment. Conclusions: Beyond the




         [page 52]                                         [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 53



                                                                       XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    indubitable protective effect against severe infection,     one line, 4 after 2 or more lines), 2 were on eltrombopag
                    our data support anti SarsCOV2 vaccination in chronic       (EPG) (after more than 2nd line), and 1 was receiving the
                    ITP patients considering the higher rate of ITP exacer-     2nd HD-DEX cycle as 1st-line treatment. The PC
                    bation observed after COVID19 infection (45,5%) than        decreased after a median of 15 days (range 1-29). Seven
                    after first (14,63%, p=0,07) and second (13.88%,            pts (58.3%) had a relapse a PC <30x109/L; the median
                    p=0,06) vaccine dose together with a higher rate of res-    PC was 20x109/L (range 2-93). Two pts (24%) had
                    cue therapy necessity in case of COVID 19 infection         mucocutaneous bleeding (grade ≤2), associated with rec-
                    (45,5% vs 7,3% and 5,5%, p=0,008 and p=0,005).              tal bleeding and metrorrhagia in 1 of them (grade 3).
                                                                                Among the pts off-treatment, six started HD-DEX, in 1
                                                                                case with HD-IVIG; among the pts on-treatment, 2 con-
                    OC075                                                       tinued EPG (N=1) or the HD-DEX cycles (N=1), and 1
                                                                                added HD-DEX to EPG. Among the 9 pts requiring treat-
                                                                                ment, the median time to reach a PC >30x109/L was 4
                    IMMUNE THROMBOCYTOPENIA DE NOVO OR
                                                                                days (range 2-6), and all pts achieved a complete
                    EXACERBATED AFTER SARS-COV-2                                response >100x109/L. The median follow-up was 24
                    VACCINATION: A MONOCENTER EXPERIENCE                        weeks (range 6-52). One patient (11.1%) relapsed after 2
                                                                                months and started EPG. Conclusions: In the last year,
                    S. Ceglie, F. Ramundo, P. Arcuri, A. Ciminello, T. Za,      one-third of ITP pts newly diagnosed at our Center was
                    S. Betti, F. Bartolomei, F. Di Landro, E. Rossi,            SARS-CoV-2 vaccination-related (15/48, 31.2%), and




                                                                                                        ly
                    V. De Stefano                                               6% of ITP pts in active follow-up experienced exacerba-
                                                                                tion after SARS-CoV-2 vaccination. However, both clin-




                                                                                                on
                    Sezione di Ematologia, Dipartimento di Scienze              ical entities had a benign course; 14/27 (51.8%) had
                    Radiologiche ed Ematologiche, Università Cattolica,         bleeding manifestations <grade 2, and only 2 pts experi-
                    Fondazione Policlinico AGemelli IRCCS, Roma, Italy          enced a bleeding grade 3. Twenty-three pts required




                                                                                           e
                                                                                treatment, and all of them reached a safe PC >30x10L
                    Background and Aims: ITP onset without accompany-
                    ing thrombosis was reported after SARS-CoV-2 vaccina-
                    tion, with an incidence of 0.8-2 and 8-11.3 per million
                                                                                    us
                                                                                within 11 days. The response was maintained in 16 of
                                                                                them (69.6%). The risk of ITP onset after vaccination is
                                                                                not predictable, but its management seems not to differ
                    doses of mRNA and ChAdOx1 vaccines. Moreover, pre-
                                                                                al
                                                                                from primary ITP.
                    existing ITP can flare up after SARS-CoV-2 vaccination
                                                                        ci

                    in about 20% of cases. Methods: From March 2021 to          Table 1.
                    March 2022, 48 ITP pts newly diagnosed were referred
                                                                  er


                    to our Center; 15 of them had de novo ITP after SARS-
                    Cov2 vaccination; moreover, 12 vaccine-related ITP
                                                            m



                    exacerbations were recorded among 200 pts on active
                    follow up. Results: Fifteen pts (M/F=9/6, median age 55
                                                 om




                    years, range 30-84) developed de novo ITP following
                    SARS-CoV-2 vaccination. Eight pts (53.3%) received an
                    adenoviral (Ad) vector-based vaccine and seven (46.7%)
                    an mRNA-based vaccine (Table 1). The platelet count
                                              -c




                    (PC) decreased after a median of 15 days (range 4-32),
                                      on




                    without a difference between vaccines. Eleven pts
                    (73.3%) had at diagnosis a PC <30x109/L; the median
                    PC was 6x109/L (range 1-38) and 17.5x109/L (range 1-
                                  N




                    32) in pts having received Ad vector-based vaccines and
                    mRNA-vaccines, respectively (p=0.66). Thirteen pts
                                                                                OC076
                    (86.7%) had bleeding manifestations, mainly mucocuta-
                    neous (N=12, grade ≤2) and genitourinary (N=2, grade 2
                    and 3). Fourteen pts required a treatment: high-dose dex-   IMMUNE THROMBOCYTOPENIA (ITP) AFTER
                    amethasone (HD-DEX) (N=12) (median 2.9 cycles) or           SARS-COV-2 VACCINATION: EXPERIENCE OF A
                    prednisone (N=2). Four cases (26.7%) received rescue        SINGLE CENTER
                    therapy with high-dose intravenous immunoglobulin
                    (HD-IVIG). The median time to reach a PC >30x109/L          F. Viola, G. Maestrini, E. Baldacci, A. Ferretti,
                    was 4 days (range 2-11), and all pts achieved a complete    R. Mormile, A. Chistolini, C. Santoro
                    response >100x109/L. The median follow-up was 35
                    weeks (range 3-59). Five pts (36%) relapsed after a medi-   Università La Sapienza, Roma, Italy
                    an of 42 days (range 36-55), requiring a 2nd-line treat-
                    ment; afterward, 3 of them received a 3rd-line treatment.   Background: Since the start of COVID-19 pandemic
                    Twelve ITP pts (M/F=3/9), median age 58.5, range 21-        there has been the urgency to develop specific vaccines.
                    79) were referred for exacerbation after Ad vector-based    ITP cases have been described after SARS-CoV-2 vac-
                    vaccines (N=1) or mRNA vaccines (N=11). Nine pts            cination. Aim of this study is to describe a monocentric
                    were off-treatment, in 2 cases after splenectomy (5 after   experience regarding the occurrence of either de novo




                                                             [Hematology Reports 2021; 13:s3]                                               [page 53]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 54



         Oral communications




                     or relapsing ITP after SARS-CoV-2 vaccination.               OC077
                     Methods: Retrospective, monocentric study. Relapse
                     definition: reduction of ≥30% in platelet (plt) count.
                     Results: A total of 17 patients (pts) experienced either     COMBINATION OF CLIA AND ELISA ASSAYS FOR
                     de novo (8) or relapsing (9) ITP, following SARS-CoV-        DIAGNOSIS OF HEPARIN-INDUCED
                     2 vaccination. De novo ITP. 8 cases, 4 females, 4 males,     THROMBOCYTOPENIA (HIT)
                     median age 66.8 years (26.2-84.1). 2 pts received
                     AstraZeneca, 1 Moderna, 5 Pfizer-BioNTech vaccine. 4         B. Montaruli, P. Valesella, M. Papandrea, A. Insana,
                     subjects were diagnosed after the 1st and 4 after the 2nd    D. Cosseddu
                     dose [median time 21.5 days (7-45)]. Median plt count
                     at diagnosis: 9x109/L (1-60); median follow up: 7.1          SC Laboratory Analysis AO Ordine Mauriziano Torino,
                     months (1.5-10.9). 6 pts presented muco-cutaneous            Italy
                     bleeding symptoms. 6/8 subjects received 1st line ther-
                     apy: 5 IvIg+corticosteroids, 1 corticosteroids. All 6 pts    Background and Aims: Heparin induced thrombocy-
                     obtained a response [median plt count 120x109/L (105-        topenia (HIT) is a serious adverse effect of heparin treat-
                     140); median time 7 days (5-39)]. Only 3/8 pts received      ment. Prompt evaluation of suspected cases is essential
                     vaccine booster after ITP development: 1 (1st dose           for clinical decisions. Aim of our study was to evaluate
                     AstraZeneca) received 2 doses of Moderna without per-        the HemosIL Acustar HIT IgG CliA cut-off for anti-
                                                                                  platelet factor 4 (PF4)/heparin antibodies of <0.13 U/ml
                     forming a blood count; 2 (ITP developed after Pfizer




                                                                                                          ly
                                                                                  suggested by Marchetti et al. in their bayesian diagnosis
                     2nd dose) received the 3rd Pfizer dose without plt count
                                                                                  of HIT. Methods: We determined in a cohort of 52 con-
                     reduction. 5/8 pts did not receive booster for their deci-




                                                                                                   on
                                                                                  secutive patients investigated for suspected HIT with a
                     sion. Relapsed ITP. 9 pts experienced an exacerbation        4T pre test probability score greater than or equal to 2
                     of ITP after either 1 or 2 vaccine doses: 5 females, 4       anti PF4-heparin IgG antibodies with CliA HemosIL
                     males, median age 48.9 years (37.5-63.6). Median num-




                                                                                              e
                                                                                  acustar HIT IgG (Werfen, USA) assay and Asserachrom
                     ber of therapies before vaccination, 1 (0-3): corticos-      anti PF4 IgG (Stago, France) ELISA assay. Patients with
                     teroids, IvIg, TPO-RA, splenectomy. Median follow up
                     6.96 years (1.13-30.75). All pts received Pfizer-
                     BioNTech vaccine. Median plt count before any of the
                                                                                      us
                                                                                  CliA assay <0.13 U/ml and ELISA assay OD <0.194
                                                                                  antiPF4 heparin antibodies were considered as negative.
                                                                                  Confirmation Platelet Aggregation Test (PAT) and
                     doses: 86x109/L (30-197); median plt count at relapse
                                                                                  al
                                                                                  HITALERT flow cytometer assays were performed in
                     after any of the doses: 26x109/L (4-48); median time at      order to confirm suspected HIT in all CliA and/or ELISA
                                                                          ci

                     relapse 7 days (1-35). At the time of 1st dose, 6 pts were   positive patients. Results: 14/52 patients were found to
                     off therapy, 2 on treatment with TPO-RA, 1 with corti-       be positive with both CliA and ELISA assays. 9/14 of
                                                                    er


                     costeroids: 2/9 relapsed (plt count 4 and 5x109/L). 1        these patients were confirmed HIT for both PAT and
                     patient presented cutaneous petechiae and was treated        HITALERT functional assays. 1/52 patient was found to
                                                              m



                     with IvIg+corticosteroids; the other one was asympto-        be positive only to CliA and 3/52 only to ELISA anti-
                     matic: corticosteroid was added to TPO-RA therapy.           PF4-heparin IgG immunoassay but negative with both
                                                  om




                     These pts obtained a response: plt count 56 and              HIT functional assays. 34/52 (63%) of the patients inves-
                     120x109/L, after 5 and 33 days, respectively. All 9 pts      tigated for suspected HIT with a 4T score=o >2 were
                     received a 2nd vaccine dose: 5 were off therapy, 3 on        found to be negative for antiPF4-heparin antibodies by
                                               -c




                     TPO-RA, 1 on corticosteroids: 6/9 relapsed (median plt       CliA assays by using literature suggested cut-off of <0.13
                     count 31x109/L; 14-48x109/L). 3 pts showed muco-             U/ml. 3 of these patients showed anti-PF4 heparin anti-
                                        on




                     cutaneous bleeding symptoms and needed therapy               bodies ELISA positivity but were not confirmed as HIT
                     (median plt count 21x109/L;14-35): 1 received corticos-      by functional assays. Conclusions: We confirmed that
                     teroids, 1 corticosteroids+TPO-RA, 1 increased corti-        the CLIA cut-off <0.13U/ml for anti-PF4.-heparin IgG
                                   N




                     costeroids dose. They all obtained a response: median        antibodies is a reasonable limit to exclude HIT in patients
                     plt count 160x109/L (42-284), median time at response        with a 4t score ≥2 or a strong HIT clinical suspicious in
                     13 days (7-34). 5/9 patients received a 3rd vaccine dose:    emergency settings. The combination of rapid CliA assay
                     3/5 were on therapy, 2 with TPO-RA, and 1 with corti-        and ELISA assay allowed us not to miss any functional
                     costeroids. 3/5 cases relapsed (median plt count             confirmed suspected HIT (see Figure 1).
                     35x109/L; 21-45), without bleeding symptoms. Just in 1
                     case therapy (corticosteroids) was necessary because of
                     low plt count (21x109/L). The patient obtained a
                     response (plt count 180x109/L), after 4 days. 4 pts did
                     not receive a third dose: 2 for patient choice, 1 for med-
                     ical decision, 1 for previous COVID-19 infection.
                     Conclusions: In our experience, few de novo or
                     relapsed ITP occurred after SARS-COV-2 vaccination,
                     with absent or mild/moderate symptoms and resolved
                     with conventional therapies or, in few cases, without
                     any treatment. These observations confirm the favor-
                     able balance towards vaccination either in pts with pre-
                     vious ITP.                                                   Figure 1.




         [page 54]                                         [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 55



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    OC078                                                       also performed to evaluate a difference in RTX rescue
                                                                                use: no significant differences were detected consider-
                                                                                ing age, median platelet count or LDH level at hospital
                    ADAMTS13 AUTOANTIBODIES AND BURDEN OF                       admission, while a statistically significant difference
                    CARE IN IMMUNE THROMBOTIC                                   was observed in terms of median ADAMTS13 inhibitor
                    THROMBOCYTOPENIC PURPURA: EVIDENCE AND                      titer at diagnosis (96.00 vs 47.00 BU/ml, p=0.015)
                    FUTURE IMPLICATIONS                                         activity. ADAMTS13 activity did not correlated with
                                                                                PEXtot, PEXtoCR nor DoH. Conclusions: From our
                    C. Dainese1, F. Valeri1, A. Valpreda2, P. Sivera3,          analysis, ADAMTS13 inhibitor titer at diagnosis
                    B. Montaruli3, M. Massaia4, B. Bruno5, A. Borchiellini1     emerges as marker of iTTP burden of care, associated
                                                                                with higher total number of PEX sessions, PEX needed
                    1Centro Riferimento Regionale per Malattie trombotiche
                                                                                to achieve clinical response, days of hospitalization,
                    ed Emorragiche dell’adulto, Ematologia Universitaria,       and a higher probability of requiring RTX rescue to
                    AOU Città della Salute e della Scienza di Torino, Italy;    achieve clinical response. Younger age, lower platelet
                    2Centro Riferimento Regionale per Malattie trombotiche
                                                                                count, higher LDH and bilirubin level at hospital
                    ed Emorragiche dell’adulto, Laboratorio Centrale, AOU       admission also show a significant correlation with total
                    Città della Salute e della Scienza di Torino, Italy;        number of PEX sessions and PEX to achieve clinical
                    3Ematologia, Ospedale Ordine Mauriziano di Torino,
                                                                                response. Despite the need for further validation,
                    Italy; 4Divisione di Ematologia, Ospedale Santa Croce       ADAMTS13 inhibitors titer could be a useful tool for




                                                                                                       ly
                    e Carle di Cuneo, Italy; 5Ematologia Universitaria,         guiding clinician defining first line treatment of new
                    Dipartimento di Oncologia, AOU Città della Salute e




                                                                                                on
                                                                                iTTP cases. Finally, such variable could be included in
                    della Scienza di Torino, Italy                              future Caplacizumab cost-efficacy analysis to better
                                                                                optimize iTTP cases stratification.
                    Background: Immune thrombotic thrombocytopenic




                                                                                            e
                    purpura (iTTP) is a rare, life-threatening thrombotic
                    microangiopathy. The introduction of a novel treatment
                    option in the acute management of iTTP,
                    Caplacizumab, has raised different questions, consider-
                                                                                    us
                                                                                al
                    ing its cost-efficacy and the optimal immunosuppres-
                    sive treatment (IST) to associate. Considering
                                                                        ci

                    Caplacizumab phase II and III trials results, in this
                    study we looked for variables associated with a higher
                                                                  er


                    burden of care and necessity of an implemented IST
                    (with early Rituximab (RTX) rescue) in order to opti-
                                                             m



                    mize resource utilization. Methods: A retrospective
                    multicenter collection of all first iTTP treated in three
                                                 om




                    different hematology units from 2007 to 2020 was con-
                    ducted. Data collected included demographic data,
                    signs and symptoms at hospital admission, laboratory
                                              -c




                    data, ADAMTS13 activity and inhibitors titer, number
                    of total PEX sessions (PEXtot) and needed to achieve        Figure 1.
                                      on




                    clinical response (PEXtoCR), days of hospitalization
                    (DoH), IST and RTX use, refractory and relapse. The
                    Mann U-Whitney test was used to assess the median           OC079
                                  N




                    differences for independent samples. Pearson correla-
                    tion coefficient (r) was applied to evaluate the correla-
                    tion among continuous variables and number of total         THROMBOTIC EVENTS WITH OR WITHOUT
                    PEX, PEXtoCR and DoH. Results: A total of 42 first          THROMBOCYTOPENIA AFTER COVID-19
                    iTTP episodes were identified. Clinical variable result-    VACCINATION IN ITALY: RESULTS FROM THE
                    ing significantly correlated with both PEXtot and           SISET VAX COVID-19 DATABASE
                    PEXtoCR were age (r=- 0.31, p=0.040 and r=-0.35,
                    p=0.030), platelet count (r=-0.30, p=0.050 and r=-0.30,     M.P. Donadini1, E. Tarasconi1, L. Bertù1, E. Antonucci2,
                    p=0.014), LDH (r=0.44, p=0.006 and r=0.41, p=0.012),        L. Contino3, V. De Stefano4, L. Russo5, G. Tiscia6,
                    total bilirubin (r=0.54, p=0.001 and r=0.35, p=0.04) and    A. Tripodi7, P. Gresele8
                    ADAMTS13 Inhibitors (r=0.46, p=0.002 and r=0.48,
                                                                                1Centro  di ricerche per le malattie tromboemboliche e
                    p=0.025). ADAMTS13 inhibitors resulted the only
                    variable associated with DoH (r=0.44, p=0.005).             le terapie antitrombotiche, Dipartimento di Medicina e
                    Considering dichotomic variables, no difference in          Chirurgia, Università dell’Insubria Varese; 2Fondazione
                    terms of median number of PEXtot and PEXtoCR and            Arianna Anticoagulazione, Bologna; 3SSD Haemostasis
                    DoH was observed considering gender, Charlson               and Thrombosis Center, Internal Medicine Department,
                    comorbididty index score or types of symptoms at hos-       “SS Antonio e Biagio” Hospital, Alessandria; 4Sezione
                    pital admission. The Mann Whitney rank sum test was         di Ematologia, Università Cattolica, Fondazione




                                                             [Hematology Reports 2021; 13:s3]                                              [page 55]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 56



         Oral communications




                     Policlinico AGemelli IRCCS, Roma; 5Division of             ing Astrazeneca or Janssen vaccines. After excluding
                     Immunohematology and Transfusion Medicine, Hospital        them (described in another report), no differences were
                     Papa Giovanni XXIII, Bergamo; 6U.O.SDipartimentale         found in terms of thrombosis location between patients
                     e di Ricerca Emostasi e Trombosi; Fondazione IRCCS         receiving mRNA and adenoviral vector-based vaccines
                     Casa Sollievo della Sofferenza, San Giovanni Rotondo;      (Table 1). Overall, during 30-day follow-up, 13 patients
                     7IRCCS Ca’ Granda Maggiore Hospital, Angelo Bianchi        (5.5%) had thrombosis progression, 10 patients (4.2%)
                     Bonomi Hemophilia and Thrombosis Center, Milano,           major bleeding and 10 patients (4.2%) died.
                     Italy; 8University of Perugia, Department of Medicine      Conclusions: Except for the very rare and peculiar
                     and Surgery – Section of Internal and Cardiovascular       cases of VITT, reported only with adenoviral vector-
                     Medicine, Italy                                            based vaccines, the clinical characteristics and out-
                                                                                comes of the remaining TEs occurring after any type of
                     Background and Aims: Since vaccine induced                 COVID-19 reflect known literature data on thrombosis
                     immune thrombotic thrombocytopenia (VITT) became           and do not differ between adenoviral vector-based and
                     evident after administration of adenoviral vector-based    mRNA vaccines. These findings seem to support recent
                     COVID-19 vaccines in early 2021, research has              epidemiological evidence of a temporal only associa-
                     focused on characterizing this new pathological entity.    tion, instead of a causal relationship, between vaccines
                     Moreover, even in absence of previously described link     and thrombotic events.
                     between vaccines and thrombosis in the pre-COVID
                     era, data were urgently needed on the association, if      Table 1.




                                                                                                        ly
                     any, between all COVID-19 vaccines and thrombosis,
                     that epidemiological data have only recently excluded.




                                                                                                on
                     Aim of the study was to collect relevant information on
                     all cases of thrombosis occurring after COVID-19 vac-
                     cination. Methods: SISET VAX COVID-19 is an ongo-




                                                                                           e
                     ing national, multicenter, prospective registry that was
                     set up by the Italian Society on Thrombosis and
                     Haemostasis, enrolling consecutive adult patients
                     objectively diagnosed with any thrombotic event (TE)
                                                                                    us
                                                                                al
                     occurring within 30 days after receiving any COVID-19
                     vaccine dose, with or without thrombocytopenia.
                                                                        ci

                     Primary objective was the characterization of TEs, in
                     terms of type and extension (venous, arterial, microvas-
                                                                  er


                     cular), diagnosis, treatment and adverse events (throm-
                     bosis progression/early recurrence, major bleeding and
                                                                                OC080
                                                             m



                     death) during 30-day follow-up. The study was
                     approved by the national ethics committee for studies
                                                 om




                     on COVID-19 and by the participating centers.              VACCINE-INDUCED THROMBOTIC
                     Fondazione Arianna Anticoagulazione provided sup-          THROMBOCYTOPENIA (VITT): EVALUATION OF
                     port for web-based dataset implementation, data man-       IMMUNOLOGIC AND FUNCTIONAL TESTS ON AN
                                              -c




                     agement and approval procedures. Results: From June        ITALIAN CASE SERIES
                     3rd, 2021, to March 20th, 2022, 237 patients from 41
                                       on




                     centers were included in the registry. The mean age was    A. Rogolino, F. Cesari, A.M. Gori, B. Giusti,
                     57 years (standard deviation (SD) 16.8) and                B. Bandinelli, E. Sticchi, A. Bertelli, M. Giannini,
                     female/male proportion was 45%/55%. TEs occurred           D. Poli, E. Lotti, L. Mannini, R. Marcucci
                                  N




                     after a mean of 14.5 days (SD 9.2) after the latest vac-
                     cine dose (72.2% after 1st or unique dose, 33.8% after     Department of Experimental and Clinical Medicine,
                     2nd dose, 1.7% after 3rd dose), that was represented by    University of Florence, Atherothrombotic Disease Unit,
                     ChAdOx1 nCOV-19 (Astrazeneca) in 28.7% of                  Azienda Ospedaliera Universitaria Careggi, Italy
                     patients, Ad26.COV2.S (Janssen) in 1.3%, BNT162b2
                     (Pfizer/BioNTech) in 54.4% and mRNA-1273                   Background and Aims: Several cases of unusual throm-
                     (Moderna) in 14.2%. 152 patients (64.1%) had at least      botic events and thrombocytopenia, called VITT (vac-
                     one comorbidity, 47 patients (19.8%) reported history      cine-induced thrombotic thrombocytopenia), have been
                     of thrombosis and 37 (17.3%) had known thrombophil-        developed after vaccination with adenoviral vectors
                     ia; 27 patients (11.4%) reported previous SARS-CoV-2       encoding the spike protein antigen of severe acute respi-
                     infection. TEs involved one or more veins in 214           ratory syndrome coronavirus 2 (SARS-CoV-2)
                     patients (90.3%) and arteries in 37 (15.6%). Among         (Vaxzevrya, AstraZeneca; Johnson&Johnson). Methods:
                     venous TE events, 170/214 (79.4%) were represented         We assessed the clinical and laboratory features of 8
                     by deep vein thrombosis and/or pulmonary embolism,         patients in Italy in whom thrombosis and thrombocy-
                     whereas 42 (19.6%) by unusual site thrombosis. In 199      topenia developed after vaccination with adenoviral vec-
                     patients (84%) the TE was not associated with thrombo-     tors-vaccines. We used three different immunologic tests
                     cytopenia. Among 38 patients (16.9%) with thrombocy-       to detect platelet factor 4 (PF4)–heparin antibodies: two
                     topenia, 22 were diagnosed with VITT, all after receiv-    standard enzyme-linked immunoadsorbent assays




         [page 56]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 57



                                                                            XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    (ELISA) (Lifecode PF4-IgG, Immucor; Zymutest HIA                 OC081
                    IgG, Hyphen), latex immunoturbidimetric assay (LIA)
                    (Hemosil HIT-Ab PF4, Werfen, Italy) and chemilumines-
                    cence immunoassay (CLIA) (Acustar IgG, Werfen,                   ANTI-SARS-COV-2 ADENOVIRAL VECTOR
                    Italy). In order to detect the functional ability of the anti-   VACCINES TRIGGER SUBCLINICAL
                    bodies in inducing platelet activation, we also performed        ANTIPLATELET AUTOIMMUNITY AND IN VIVO
                    two different functional tests: Heparin-induced platelet         PLATELET ACTIVATION
                    Activation-HIPA test modified by the addition of PF4 on
                    washed platelets and flow cytometry (FC) on platelet rich        E. Petito1, E. Colonna1, E. Falcinelli1,
                    plasma. Results: All patients [2M/6F; median age: 66             A.M. Mezzasoma1, E. Cesari1, E. Giglio1, T. Fiordi2,
                    (41-78)] had a confirmed diagnosis of VITT based on              F. Almerigogna3, A. Villa4, P. Gresele1
                    clinical and laboratory features. Blood samples were             1Section
                    obtained, except cases #1,#2,#6,#8 before i.v.                            of Internal and Cardiovascular Medicine, and
                                                                                     2Section  of Occupational Medicine and Toxicology,
                    Immunoglobulin (IG) or anticoagulant administration.
                    As shown in the Table 1, there was a significant correla-        University of Perugia, Perugia, Italy; 3Unit of
                    tion between the two different ELISA tests (rho=0.683,           Allergology and Clinical Immunology, Department of
                    p=0.042). According to the ELISA cut-off values (>0.4            Clinical and Experimental Medicine, University of
                    for Immucor and >0.3 for Hyphen) all patients were pos-          Florence, Florence, Italy; 4Central Clinical Chemistry
                    itive for the presence of anti-PF4 antibodies. None of the       Laboratory, S.Mdella Misericordia Hospital, Perugia,




                                                                                                             ly
                    patients had LIA and CLIA values above the cut-off (>1           Italy




                                                                                                      on
                    U/mL). By performing functional tests, all VITT patients
                    resulted positive to the functional modified HIPA test,          Background and Aims: To slow down the COVID-19
                    except for VITT #6 patient, who resulted negative and            pandemic, an unequalled vaccination campaign against
                    VITT #2 patients who resulted weakly positive, probably          SARS-CoV-2 has been started all around the world




                                                                                                e
                    due to the concomitant IG therapy. By performing FC              (PMID: 35491495). While a few studies have evaluated
                    functional test, VITT #1, #2, #6, #8, with the concomi-
                    tant IG therapy showed a significant lower percentage of
                    activated platelets (mean fluorescence intensity (MFI) of
                                                                                         us
                                                                                     the incidence of anti-PF4 positivity and platelet activa-
                                                                                     tion in anti-SARS-CoV-2 vaccine recipients (PMID:
                                                                                     33988688, PMID: 34246010, PMID: 34887867), no
                                                                                     al
                    CD62P platelets) with respect to the VITT patients with-         studies have assessed whether an immunological
                    out IG therapy [MFI: 4458 (4320-4629) vs 11170 (6716-            response involving platelets develops in healthy subjects
                                                                            ci

                    17492), p=0.03]. Patients #4F, #5F, showed a great               (HS) after anti-SARS-CoV-2 vaccination and if this asso-
                    platelet activation with VITT serum and buffer, and a            ciates with platelet activation. We carried out a prospec-
                                                                      er


                    decreased CD62P expression with low dose of unfrac-              tive study in HS who received the first dose of adenovi-
                    tionated heparin(UFH) (0.3 IU/mL), by showing a typi-            ral-vector vaccines (ChAdOx1 or Ad26.COV2.S) or
                                                                m



                    cal positive VITT pattern. In patients #7, we found an           mRNA vaccine BNT162b2 to evaluate platelet-specific
                    enhanced CD62P expression in the tube with unfrac-               immune response and in vivo platelet activation.
                                                   om




                    tioned heparin 0.3 IU/mL, with a typical HIT pattern.            Methods: 72 HS undergoing first dose ChAdOx1
                    Conclusions: Vaccination with adenoviral vectors-vac-            (n=32), Ad26.COV2.S (n=20) or BNT162b2 (n=20)
                    cines encoding the spike protein antigen of SARS-CoV-            administration were enrolled and studied at two time
                                                -c




                    2 can result in the rare development of VITT. Our data           points: on the day of vaccination (T0), just before vac-
                    indicate that the ELISA test in association with function-       cine administration, and after 13.9±0.4 days (T1).
                                        on




                    al test assay are able to detect the presence of platelet        Peripheral venous blood was collected in trisodium cit-
                    activating antibodies against PF4 in VITT. HIPA test             rate 3.2% (0.109 M, 1/10 v/v), 0.18% K3EDTA and non-
                    remains the gold standard to detect functional ability of        anticoagulated glass tubes. Anti-PF4/heparin antibodies
                                   N




                    antibodies anti PF4 to activate platelets. In addiction the      were searched in serum by two different ELISA; their
                    FC seems a sensitive and reliable method with respect to         ability to induce platelet activation was assessed by flow-
                    HIPA, however the evaluation in sera of patients treated         cytometry; antiplatelet auto- and allo-antibodies were
                    with IG might be associated with false negative results          searched in serum by ELISA; platelet-derived micropar-
                                                                                     ticles (PMPs) were measured in platelet-free plasma by
                    and in some cases it results less sensitive with respect to
                                                                                     flow cytometry; plasma levels of sP-sel were measured
                    HIPA test in detecting platelet activating antibodies.
                                                                                     by ELISA.Results: Anti-PF4/heparin antibody positivity
                    Table 1.                                                         after vaccination was relatively infrequent and compara-
                                                                                     ble in the three groups, and when present it was at low
                                                                                     titer and not associated with thrombocytopenia or with
                                                                                     platelet activation. On the other hand, adenoviral vector
                                                                                     vaccine-elicited antiplatelet auto- (34.3% in ChAdOx1
                                                                                     and 25% in Ad26.COV2.S) and allo-antibody positivity
                                                                                     (18.7% in ChAdOx1 and 30% in Ad26.COV2.S).
                                                                                     Antiplatelet auto- or allo-antibody positivity pattern was
                                                                                     not specific, but rather an unspecific immune response
                                                                                     against platelet antigens. In the ChAdOx1 group, circu-
                                                                                     lating PMPs and s-Psel increased significantly at T1




                                                                 [Hematology Reports 2021; 13:s3]                                                  [page 57]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 58



         Oral communications




                     compared with T0 (Figure 1). Moreover, subjects who                ADAMTS13 activity were stable among the three time-
                     became positive for antiplatelet auto- or allo-antibodies          points (Figure 1). In only two patients a significant drop
                     after ChAdOx1 vaccination displayed a significantly                in ADAMTS13 levels occurred after the first dose (from
                     higher number of circulating PMPs compared to those                28% to <3% and from 101% to 82%), and both remained
                     who remained negative. Conclusions: Our study shows                stable after the second dose, with negative anti-
                     that an immunological reaction involving platelets is not          ADAMTS13 antibodies. Due to a stable undetectable
                     uncommon in subjects receiving anti-SARS-CoV-2 vac-                ADAMTS13, the first patient was treated with 4 doses of
                     cination, especially after ChAdOx1 and, although less              weekly 375 mg/m2 rituximab with a rapid ADAMTS13
                     commonly, with Ad26.COV2.S, and that it associates                 response. One patient had positive basal anti-
                     with in vivo platelet activation, suggesting that a deregu-        ADAMTS13 antibodies with a titer remaining stable
                     lated immune response involving platelets following                after the two vaccine doses, while in another patient anti-
                     adenoviral-based vector vaccination occurring in some              ADAMTS13 antibodies became detectable after the first
                     subjects triggers platelet activation. Given the absence of        dose, with no corresponding drop in ADAMTS13 levels
                     vaccine complications in enrolled subjects, our data               and a stable titer after the second dose. Conclusions: The
                     show that these alterations do not have a predictive value         result of our study prospectively evaluating the effect of
                     for VITT, but it is conceivable that this response may rep-        anti-SARS-CoV-2 vaccination on the risk of relapse in a
                     resent a priming trigger which, in association with some           large cohort of patients with TTP in Milan showed a
                     yet unknown rare individual predisposition, may lead to            lower than reported relapse rate (1.36% vs 2.6%) with an
                     full blow VITT in a few subjects.                                  observed to expected incidence rate ratio of 0.52, con-




                                                                                                                ly
                                                                                        firming the safety of mRNA-based anti-SARS-CoV-2




                                                                                                         on
                                                                                        vaccination in TTP patients.


                                                                                        OC083




                                                                                                   e
                     Figure 1. Platelet activation. (A) Plasma levels of PMPs and (B)
                     Soluble platelet P-selectin in subjects undergoing first dose of
                                                                                            us
                                                                                        VACCINE INDUCED THROMBOTIC
                                                                                        THROMBOCYTOPENIA: EVALUATION OF GENETIC
                     ChAdOx1 or BNT162b2, or unique dose of Ad26.COV2.S on              SUSCEPTIBILITY THROUGHT WHOLE EXOME
                                                                                        al
                     the day of vaccination (T0) and after 13.9±0.4 days (T1).          SEQUENCING
                                                                               ci

                                                                                        T. Capezzuoli1, R. De Cario1, E. Sticchi2, M. Giannini1,
                                                                         er


                     OC082                                                              R. Orsi1, L. Squillantini1, S. Suraci1, A.M. Gori2,
                                                                                        R. Marcucci2, B. Giusti2
                                                                   m



                     IMPACT OF SARS-COV2 VACCINATION IN
                                                                                        1University of Florence, Department of Experimental
                     THROMBOTIC THROMBOCYTOPENIC PURPURA
                                                     om




                     PATIENTS                                                           and Clinical Medicine, Florence; 2University of
                                                                                        Florence, Department of Experimental and Clinical
                     A. Artoni, M. Capecchi, P. De Leo, M. Biganzoli,                   Medicine, Atherothrombotic Diseases Center, Careggi
                                                  -c




                     I. Mancini, P. Agosti, B. Ferrari, R. Gualtierotti,                Hospital, Florence, Italy
                     F. Peyvandi, A. Bianchi Bonomi
                                          on




                                                                                        Background: In late February 2021, a prothrombotic
                     Hemophilia and Thrombosis Center, Fondazione IRCCS                 syndrome was observed in a small number of individuals
                     Ca' Granda Ospedale Maggiore Policlinico, Milano,                  who received the adenoviral vector-based vaccine
                                    N




                     Italy                                                              Vaxzevria (AstraZeneca). Similar pictures were observed
                                                                                        in some individuals who received the Janssen (Johnson
                     Background: Thrombotic thrombocytopenic purpura                    & Johnson) vaccine. This syndrome has been named vac-
                     (TTP) has occasionally been described after vaccination.           cine-induced thrombotic thrombocytopenia (VITT) and
                     Since the availability of anti-SARS-CoV-2 vaccines, 12             is characterized by multiple venous/arterial thrombosis,
                     cases have been described on a possible association with           with atypical venous thrombosis of cerebral and splanch-
                     TTP onset. Aims: This study aims to evaluate the relapse           nic districts, associated with thrombocytopenia.
                     rates in patients affected by TTP undergoing anti-SARS-            Hemorrhagic manifestations and sometimes disseminat-
                     CoV-2 vaccination. Methods: All consecutive TTP                    ed intravascular coagulation (DIC) are present.
                     patients undergoing anti-SARS-CoV-2 vaccination from               Evidences suggest that this rare syndrome is caused by
                     March to May 2021 were enrolled. Blood samples were                platelet-activating antibodies directed against platelet
                     collected before vaccination (T0), 2 weeks after the first         factor 4 (PF4). Aim of this work is to apply a Whole
                     (T1) and the second dose (T2) to evaluate ADAMTS13                 Exome Sequencing (WES) analysis approach, for the
                     activity and anti-ADAMTS13 antibody titer. Results: A              identification of possible genetic predisposition profiles
                     total of 49 TTP patients were enrolled (48 acquired and 1          underlying VITT. Methods: Due to the referral role of
                     congenital), all vaccinated with an mRNA vaccine. No               the Center for Atherothrombotic Diseases (University of
                     patients had a clinical TTP relapse, with an ADAMTS13              Florence/AOU Careggi) for the management of COVID-
                     relapse rate of 1.36% per month. Mean levels of                    19 patients and diagnosis of VITT, fifty patients were




         [page 58]                                             [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 59



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    examined. Ten out of fifty patients were diagnosed with       thrombotic events, and other autoimmune adverse reac-
                    VITT. In our analysis we used Next Generation                 tions have been reported in connection with some of the
                    Sequencing techniques, by a WES approach, with                SARS-CoV-2 vaccines, and have caused a lot of worry
                    Illumina NextSeq500 platform and SureSelect XT HS             and a severe confusion within the population and the
                    enrichment kit (Agilent Technologies), to analyse six out     medical community. Antithrombin deficiency is recog-
                    ten VITT patients. Results: VITT patients analysed were       nized as the most severe form of inherited thrombophil-
                    6 females (mean age 64,2±13,8), who received an aden-         ia. We assessed the risk of thrombotic events in carriers
                    oviral vector-based vaccine. WES analysis revealed a          of antithombin deficiency after SARS-CoV-2 vaccina-
                    total of 140,563 variants. Rare variants (MAF <1%)            tion. Methods: We estimated the absolute risk of
                    identified range from 1,619 to 1,774 and their distribu-      thrombosis within 30 days of each single dose of 3
                    tion by type (frameshift, missense, splicing, nonsense,       SARS-CoV-2          vaccinations       [    ChAdOx1-S
                    UTR, samesense, intronic) is similar in the six patients.     (AstraZeneca), COVID-19 Vaccine Moderna mRNA -
                    In this work we decided to focus on rare variants             1273      (Spikevax),       Pfizer    mRNABNT162b2
                    involved in different biological processes underlying         (Comirnaty)] in carriers of antithrombin deficiency,
                    VITT. We found a total of 89 rare variants in genes           compared with non carriers. Results: In this prospec-
                    involved in integrin signaling pathways (ITGA2B,              tive family cohort study, on the basis of the pedigree
                    ITGAD, ITGB4, GP6, FGA, FGB), in thrombocytopenia             analysis, we enrolled a total of 141 subjects (47% men;
                    (MASTL, PDIA6, FYB, MYH9), and other genes induc-             median age 57 years), belonging to 21 different families
                    ing/inhibiting platelet aggregation/activation processes.     with inherited deficiency of antithrombin. Out of 141




                                                                                                          ly
                    Interestingly, the two patients (VITT05 and VITT18),          subjects, 81 (57%; 47% men) were carriers of




                                                                                                   on
                    with most severe clinical complications, showed a higher      antithrombin deficiency and 60 (43%; 48% men) were
                    number of rare variants identified in such pathways (21       not. In the group of defect carriers 68 received three
                    and 27 variants, respectively). Among the abovemen-           vaccine doses, 6 two doses and 2 received 1 dose; 3 car-
                    tioned pathways, 15 variants with putative functional         riers refused vaccination. In the non-carrier group 55




                                                                                             e
                    effect have been identified in genes encoding for inte-       received three vaccine doses, and one two doses; 4 non-
                    grins or integrin ligands, as well as in molecules involved
                    in pathways triggered by these molecules. Interestingly,
                    both VITT05 and VITT18, patients with a more severe
                                                                                      us
                                                                                  carriers refused vaccination. A total of 220 (172
                                                                                  Comirnaty, 32 Spikevax, 16 ChAdOx1-S) and 167 (109
                                                                                  Comirnaty,48 Spikevax, 10 ChAdOx1-S) vaccine doses
                                                                                  al
                    phenotype, carried variants in GP6 gene, encoding a col-      were administered in carriers and non-carriers, respec-
                    lagen receptor involved in collagen-induced platelet          tively. Thirty-five carriers of the defect were taking
                                                                          ci

                    adhesion and activation. Conclusions: WES analysis            anticoagulant therapy for previous thrombotic events.
                    exhibit a considerable number of variants in molecular        Only one thrombotic event occurred in a 44-year-old
                                                                    er


                    pathways involved in integrin signaling, thrombocytope-       woman with the defect two weeks after receiving the
                    nia and platelet aggregation/activation processes The two     second dose of the vaccine. The relative risk was of
                                                              m



                    patients with the worst clinical outcome presented a sig-     0.75 (95% CI 0.05 to 11.8); even when anticoagulated
                    nificantly higher number of suggestive rare variants with     patients were excluded the relative risk was of 1.3 (95%
                                                  om




                    respect to other patients investigated; consequently, it is   CI 0.08 to 20). Six carriers developed covid disease
                    not possible to exclude the possible contribution of a        (one before vaccination, one after the first dose, 2 after
                    greater number of rare suggestive variants in the modu-       the second, 2 after the third) versus two in the non-car-
                                               -c




                    lation of the phenotype of patients with worse clinical       rier group (after the third dose) : RR 2.14 (95% CI 0.5
                    course. Further investigation on other mechanisms             to 10.2). Conclusions: The data of our study show that
                                       on




                    (inflammation, immunity, viral response) and functional       even in patients genetically predisposed to an increased
                    assays are needed for more clarity with respect to the        thrombotic risk, vaccination against Covid does not
                    impact of genetic background on VITT susceptibility.          seem to increase this risk.
                                  N




                    OC084                                                         OC085

                    THE RISK OF DEVELOPING THROMBOTIC EVENTS                      HEMOSTATIC BIOMARKERS AND SEROLOGICAL
                    IN INDIVIDUALS WITH ANTITHROMBIN                              RESPONSE IN SUBJECTS RECEIVING ANTI-
                    DEFICIENCY UNDERGOING COVID VACCINATION:                      COVID-19 VACCINATION: A PROSPECTIVE
                    A FAMILY COHORT STUDY                                         COHORT STUDY

                    D. Tormene, E. Bicego, N. Perin, S. Gavasso, P. Simioni       F. Schieppati1, S. Gamba1, E. Galimberti1, L. Russo1,
                                                                                  C. Giaccherini1, S. Bolognini1, C.J. Tartari1, C. Ticozzi1,
                    Thrombotic and Haemorrhagic Diseases Unit,                    A.M. Palladino1, O.C. Cretu1, L. Barcella1,
                    Department of Medicine, University of Padua Medical           M. Marchetti1, A. Falanga1,2
                    School, Padua, Italy
                                                                                  1Immunohematology       and Transfusion Medicine,
                    Background and Aims: After SARS-CoV-2 vaccina-                Hospital Papa Giovanni XXIII, Bergamo; 2Medicine and
                    tion campaign initiation, rare, unusual, and severe           Surgery, University of Milano Bicocca, Milano, Italy




                                                               [Hematology Reports 2021; 13:s3]                                                 [page 59]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 60



         Oral communications




                     Background and Aims: SARS-COV2 infection is asso-             fibrinolysis activation. Compared with recent studies, our
                     ciated with inflammation, hypercoagulability and              results provide a longer observation follow-up with all
                     endothelial damage. Anti-SARS-COV2 vaccines have              vaccine types and reassure on the safety of anti-SARS-
                     radically changed the course of the pandemic, although        COV2 vaccination.
                     reports on rare thrombotic adverse events have raise con-
                     cern in the scientific and general communities. We
                     prospectively enrolled a cohort of adult subjects under-      OC086
                     going mRNA or adenovirus vector vaccination, with the
                     aim to longitudinally evaluate: 1. changes in hemostatic
                                                                                   NATURAL HISTORY OH ANTI PFA ANTIBODIES IN
                     biomarkers (i.e. clotting activation, endothelium and fib-
                     rinolysis); 2. the serological response; 3. the occurrence    PATIENTS WITH VITT
                     of manifest thrombotic complications. Methods:
                     Peripheral blood samples were collected at enrollment         E. Lotti, F. Vannini, A. Rogolino, F. Cesari, A. Bertelli,
                     (day 0, D0) before the 1st vaccine dose, and on 15 (D15),     L. Mannini, D. Poli, B. Giusti, R. Marcucci
                     60 (D60), 90 (D90) and 180 (D180) days after the 1st
                     dose. At each time point, hemostatic markers (i.e., fib-      Department of Experimental and Clinical Medicine,
                     rinogen, D-dimer, FVIII, von Willebrand Factor [vWF]          University of Florence; Atherothrombotic center, AOU
                     antigen and activity, F1+2, thrombomodulin, protein C,        Careggi, Italy
                     protein S, FXIII, tPA, and PAI-1), and anti-Spike recep-




                                                                                                            ly
                     tor-binding-domain protein (anti-S/RBD) IgG were              Background and Aims: Vaccine-induced Immune
                     measured. Results: Fifty-three subjects (57% males,           Thrombocytopenia and Thrombosis (VITT) is a new




                                                                                                    on
                     median age 50 years [range 23-86]) were enrolled into         clinical syndrome discovered during the first half of
                     the study and followed-up for 180 days: 36 (68%)              2021, when very rare cases of thrombocytopenia com-
                     received BNT162b2, 6 (11%) mRNA-1273, and 8 (15%)             bined with unusual thrombosis (venous and/or arterial),




                                                                                              e
                     ChAdOx1 nCoV-19 vaccines, in 2 doses over 21, 30 and          have been reported one to two week after vaccination
                     77 days, respectively; 3 (6%) subjects received
                     Ad26.COV2.S as single shot. A previous history of
                     COVID-19 was reported by 20 individuals (38%), with a
                                                                                       us
                                                                                   with Adenovirus-based vaccines against SARS-CoV-2.
                                                                                   VITT seems to share the same pathogenetic mechanisms
                                                                                   of autoimmune heparin-induced thrombocytopenia
                                                                                   (aHIT); specifically, both VITT- and aHIT-patients
                                                                                   al
                     mean of 10 months (4-18) before vaccination; only 1
                     required hospitalization. Nine subjects presented cardio-     develop an high serum titer of IgG antibodies directed
                                                                           ci

                     vascular risk factors and 4 a prior, non-active, cancer; 3    against platelet-factor-4 (PF4) even in the absence of
                     were on anticoagulation for atrial fibrillation. The evalu-   heparin exposure. Initial data on the natural history of
                                                                     er


                     ation of the hemostatic biomarkers at the different time      antiPF4 antibodies in large VITT-patients cohorts have
                     points showed variations in some of the parameters eval-      been recently published suggesting a longer antiPF4 titer
                                                               m



                     uated, with median values remaining within normal             durability compared to what has been previously
                     range levels. Specifically, compared to baseline, we          described in HIT patients. Here we present the results of
                                                   om




                     observed a significant increase in thrombomodulin at          an ongoing study on a VITT-patients cohort followed
                     D90 (p=0.001) and D180 (p=0.03), and a significant            with serial determinations of quantitative serum antiPF4
                     decrease in fibrinogen (D60), vWF-Ag (D60 and D180),          antibodies titer and functional HIPA testing for a median
                                                                                   follow-up duration of 6 months. Methods: We included
                                                -c




                     FVIII (D60, D90 and D180), and TPA (D60 and D90)
                     levels. The reduction of these biomarkers was particular-     nine patients (seven females and three males) with a
                                                                                   diagnosis of VITT confirmed between March and June
                                        on




                     ly evident in individuals with a history of COVID-19. Of
                     interest, this group of subjects was also characterized by    2021. The median age at the time of diagnosis was 63,5
                     significantly lower levels of PAI-1 both at baseline (7.18    years. All patients developed thrombocytopenia with a
                                   N




                     ng/mL vs 17.53 ng/mL; p<0.0001), and at other time            median platelet count at the nadir of 23.500/mmc
                     points (p<0.0001), and by an increase in F1+2 at D90          (10.000-95.000). All developed thrombotic manifesta-
                     (p=0.02). The association between lower baseline PAI-1        tions with a median D-Dimer level at the presentation of
                     levels with history of COVID-19 was confirmed by lin-         28330 ng/ml (5.151-74.124). The overall mortality rate
                     ear regression analysis (B=-10.351, p=0.013), and was         in this cohort was 20% (2/9), whereas another 20% (2/9)
                     independent by the time of infection resolution. Notably,     of patients survived developing moderate to severe dis-
                     no differences were observed in the hemostatic biomark-       ability. To detect platelet factor 4 (PF4)–heparin antibod-
                     ers according to vaccine types. All subjects positively       ies we used a standard enzyme-linked immunoadsorbent
                     responded to vaccination with a significant increase in       assay (ELISA) (Lifecode PF4-IgG test, Immucor, Itay)
                     anti-S/RBD IgG from baseline (D0) to each time point,         and a chemiluminescence immunoassay (CLIA)
                     especially COVID-19 subjects (D15, D60, and                   (Acustar IgG test, Werfen, Italy). A functional test
                     D90:p<0.0001; D180:p=0.031). No thrombotic or car-            Heparin-induced platelet Activation (HIPA) test modi-
                     diovascular events occurred during follow-up.                 fied by the addition of PF4 in order to detect the function-
                     Conclusions: We found no hypercoagulable state elicited       al ability of the antibodies in inducing platelet activation
                     by COVID-19 vaccination, contrarily we detected an            was performed (Figure 1). Results: At the time of diag-
                     overall persistent reduction of clotting activation over      nosis the median antiPF4 antibodies quantitative titer
                     time. Subjects with previous SARS-COV2 infection had          was extremely high (2472 OD units, range), associated in
                     persistently low levels of PAI-1, supporting enhanced         all cases with a positive functional test. In the remanining




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    survivors cohort (7 patients), 3 patients remained positive   sion (i) observational studies on TTS providing results
                    at the quantitative assay after a median follow-up dura-      of the performed laboratory diagnostic tests, (ii) studies
                    tion of 6 months. In one of these three patients we regis-    investigating the prevalence of anti-PF4 antibodies in
                    tered the maximum durability of positive antiPF4 of 9         vaccinees with an adenoviral vector vaccine with no
                    months, with a value of 1176 OD units with a persistent       signs or symptoms of TTS, and (iii) laboratory studies
                    functional positivity. Interestingly in another patient we    investigating novel diagnostic tests and/or comparing
                    found a still detectable functional positivity after almost   different available diagnostic tests.
                    1 year from diagnosis, despite seroconversion to negative
                    quantitative assay occurred after 4 months. None of the       Table 1.
                    surviving patients included in this study showed clinical
                    manifestations of relapse. Conclusions: For the first time
                    in a real life study we have documented in patients with
                    diagnosis of VITT the natural history of anti-PFA IgG
                    antibodies at long follow up intervals (up to 12months)
                    both by immunological and functional assays.




                                                                                                          ly
                                                                                                  on
                                                                                             e
                                                                                      us
                                                                                  al
                                                                          ci
                                                                    er


                    Figure 1.
                                                              m




                    OC087
                                                  om




                    SENSITIVITY AND SPECIFICITY OF DIAGNOSTIC
                    TESTS FOR THROMBOSIS WITH
                                               -c




                    THROMBOCYTOPENIA SYNDROME -
                    A SYSTEMATIC REVIEW OF CASE REPORTS AND
                                       on




                    CASE SERIES
                                  N




                    B. Clerici1, M. Scavone1, S. Birocchi1, C. Aloise1,
                    B. Peroni1, A. Negrini1, G. Casazza2, G.M. Podda1
                    1Divisione di Medicina Generale II, ASST Santi Paolo e
                    Carlo, Dipartimento di Scienze della Salute, Università
                    degli Studi di Milano, Milan, Italy; 2Dipartimento di
                    Scienze Biomediche e Cliniche “L. Sacco”, Università
                    degli Studi di Milano, Milan, Italy

                    Background and Aims: Diagnostic accuracy measures
                    of the available diagnostic tests for Thrombosis with
                    Thrombocytopenia Syndrome (TTS) are presently lack-
                    ing. The aim of the study was to estimate sensitivity and
                    specificity of PF4-ELISAs, Platelet Activation Assays
                    (PAAs) and PF4-enhanced PAAs (PF4-PAAs).
                    Methods: A systematic search of the literature was per-
                    formed by 2 researchers with a pre-defined research
                    string and a snowball strategy. We evaluated for inclu-       Patients were considered affected by TTS according to




                                                               [Hematology Reports 2021; 13:s3]                                                [page 61]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 62



         Oral communications




                     the definition of the authors or guidelines. Results: Fifty-    more affected by peripheral obstructive arterial disease
                     seven studies describing diagnostic test results of 508         (POAD) and atrial fibrillation (AF), and presented a his-
                     TTS patients and 691 vaccinees with adenoviral vector           tory of previous MB, with respect to patients who did not
                     vaccines without signs or symptoms of TTS were includ-          bleed. Patients with MB showed a trend for lower time in
                     ed in the analysis. The summary sensitivity estimates of        therapeutic range (TTR), and a significant number of
                     PF4-ELISAs, PAAs and PF4-PAAs were 94.5% (95%CI,                patients had a TTR in the lower quartile. Patients with
                     92.4-96.1%), 65.1% (95%CI, 55.2-74.1%), and 96.7%               MB had a higher mortality rate with respect to patients
                     (95%CI, 90.6-99.3%), respectively (Table 1, Panel A).           who did not bleed (p=0.001). The history of previous
                     Summary estimates of specificity were 95.5% (95%CI              bleeding, the presence of POAD or of AF, and a TTR in
                     93.7-96.9%) for PF4-ELISAs and 100% (95%CI, 92.9-               the lowest quartile, were significantly associated with
                     100.0%) for PF4-PAAs; data for specificity estimation of        MB. Conclusions: MHV patients treated with VKAs fol-
                     PAAs were scarce (Table 1, Panel B). Conclusions: Our           lowed by Anticoagulation Clinics, showed a low bleed-
                     results suggest that PF4-ELISAs, due to high sensitivity        ing risk. Risk factors associated with major bleeding are
                     and high specificity, constitute appropriate tests to rule in   older age, the presence of POAD or AF, the history of
                     and rule out TTS. PF4-PAAs, highly sensitive and specif-        previous bleeding, and poor quality of anticoagulation.
                     ic tests limited by their low availability and technical        Patients who experienced MB during anticoagulation are
                     complexity, can be used in selected cases to confirm or         at high risk of death.
                     exclude TTS.




                                                                                                            ly
                                                                                     OC089




                                                                                                     on
                     OC088
                                                                                     DOACS USE IN EXTREME BODY WEIGHTED
                     MAJOR BLEEDINGS IN MECHANICAL                                   PATIENTS: RESULTS FROM THE PROSPECTIVE




                                                                                               e
                     PROSTHETIC HEART VALVES PATIENTS ON                             START-REGISTER
                     VITAMIN K ANTAGONIST TREATMENT. DATA
                     FROM THE PLECTRUM STUDY                                            us
                                                                                     M. Guarascio1, L. Bertù2, M.P. Donadini2,3,
                                                                                     E. Antonucci4, G. Palareti4, W. Ageno2,3
                     D. Poli1, E. Antonucci2, R. Facchinetti3, P. Falco4,
                                                                                     al
                                                                                     1U.OPneumologia 2Covid, “Vito Fazzi” Hospital,
                     G. Serricchio5, T. Lerede6, L. Masciocco7, P. Gresele8,
                     G. Palareti2, S. Testa9 on the behalf of the Italian            Lecce, Italy; 2Research Center on Thromboembolic
                                                                             ci

                     Federation of Anticoagulation Clinics                           diseases and Antithrombotic Therapies, University of
                                                                      er


                                                                                     Insubria, Varese, Italy; 3Emergency Medicine and
                     1Center   of Atherothrombotic Disease, Azienda                  Thrombosis and Haemostasis Center, ASST Sette Laghi,
                                                                m



                     Ospedaliero-Universitaria Careggi, Firenze; 2Fondazione         Varese, Italy; 4Arianna Anticoagulazione Foundation,
                     Arianna Anticoagulazione, Bologna; 3Laboratorio                 Bologna, Italy
                                                    om




                     Analisi, Azienda Ospedaliera Universitaria Integrata
                     (AOUI) Ospedale Civile Maggiore Di Borgo Trento;                Background and Aims: Direct oral anticoagulants
                     Verona; 4Poliambulatorio Specialistico MEDICAL                  (DOACs) are widely used for the treatment of venous
                     PONTINO, Latina 5Lab analisi Ospedale Sant’Anna San             thromboembolism (VTE) and for stroke prevention in
                                                 -c




                     Fermo della Battaglia, ASST Lariana Como; 6Centro               atrial fibrillation (AF). However, evidence in obese and
                                         on




                     Trombosi e Emostasi Immunoematologia e Medicina                 underweight patients is limited. We assessed the safety
                     Trasfusionale ASST Papa Giovanni XXIII Bergamo;                 and effectiveness of DOACs and vitamin K antagonists
                     7UOC Medicina Interna, Ospedale Lastaria, Lucera                (VKAs) in patients >120 Kg or <50 Kg enrolled in an
                                    N




                     (FG); 8Department of Medicine, Section of Internal and          observational prospective cohort study, the START-
                     Cardiovascular Medicine, University of Perugia;                 Register. Methods: Adult patients started on anticoagu-
                     9Department of Laboratory Medicine, Haemostasis and             lant therapy were followed up for a median of 1.5 years
                     Thrombosis Centre, ASST Cremona, Cremona, Italy                 (IQR 0.6-2.8). Primary efficacy outcome was the occur-
                                                                                     rence of VTE recurrence, stroke and systemic embolism.
                     Background and Aims: Patients with mechanical pros-             Primary safety outcome was major bleeding (MB)
                     thetic heart valves (MHV) need vitamin K antagonist             Results: 10,080 AF and VTE patients were enrolled
                     (VKA) treatment, due to the high thrombotic risk. The           between March 2011 and June 2021, 295 patients
                     need to evaluate the bleeding risk of these patients is of      weighted <50 Kg and 82 patients >120 Kg. Obese
                     great clinical relevance. Methods: Observational retro-         patients were significantly younger than underweight
                     spective multicenter study among Centers affiliated to          patients. Rates of thrombotic events were low and similar
                     the Italian Federation of Anticoagulation Clinics (FCSA)        between DOACs and VKAs in underweight patients (1
                     on MHV patients, with the aim to evaluate the risk of           event on DOACs therapy [0.9% 95%CI 0.11-5.39] and 2
                     major bleeding (MB) and associated risk factors.                on VKAs [1.1% 95%CI 0.01-47.68]) and in overweight
                     Results: 2357 patients with MHV were included in the            patients (0 events on DOACs, 1 on VKAs [1.6%, 95%CI
                     study, 246 patients had Major Bleeding (MB) (rate1.0            0.11-5.79]. Two MB events occurred on DOACs (1.9%,
                     x100 pt-yrs), 54 were intracranial hemorrhage (rate 0.22        95%CI 0.38-6.00) and 3 on VKAs (1.6%, 95%CI 0.04-
                     x100 pt-yrs). Patients with MB were significantly older,        22.06) in the underweight group; 1 MB on DOACs




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 63



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    (5.3% 95%CI 0.33-16.68) and 2 on VKAs (3.3%, 95%CI             discharge and after 6 months were recorded on a web-
                    0.02-130.77) in the overweight group. (Table 1).               based case report form. To define the major bleeding we
                    Conclusions: DOACs seem to be effective and safe also          adopted the criteria of the International Society on
                    for the treatment of patients with extreme body weights,       Thrombosis and Haemostasis. Results: Between January
                    both underweight and overweight. Further prospective           2015 and December 2021, 224 patients with major bleed-
                    studies are needed to support these findings.                  ing events were enrolled. Non-valvular atrial fbrillation
                                                                                   (NVAF) was the indication for treatment in 86% of
                    Table 1.                                                       patients; 59% were males; median age 80 years; 31.7% of
                                                                                   patients were on apixaban; 21.9% on dabigatran,7.1% on
                                                                                   edoxaban; 39.3% on rivaroxaban. An inappropriate dose
                                                                                   of DOAC were administered in 41 patients (18%).
                                                                                   Concomitant antiplatelet treatment was present in 14% of
                                                                                   enrolled patients; 111 patients (49.5%) had intracranial
                                                                                   bleeding (ICH), 11 of them were fatal, 83 (37.1%) patients
                                                                                   had gastrointestinal bleedin (GIB), 6 were fatal; 30
                                                                                   (13.4%) patients had major bleeding in other site, none
                                                                                   was fatal. Therapeutic interventions for the management
                                                                                   of bleeding were performed in about 90% of patients.
                                                                                   Only in 10% of patients a conservative approach was




                                                                                                            ly
                                                                                   recorded, expecially in patients enrolled within 2015 and
                    OC090
                                                                                   2018.Therapeutic strategies with/without surgery or inva-




                                                                                                    on
                                                                                   sive procedures included: fluid replacement or red blood
                    MANAGEMENT OF MAJOR BLEEDING IN                                cells transfusion, prothrombin complex concentrates (3 or
                    PATIENTS TREATED WITH DIRECT ORAL                              4 factors), antifbrinolytic drugs, and the administration of




                                                                                              e
                    ANTICOAGULANTS: UPDATE DATA FROM START                         specific reversal strategies. In particular idarucizumab was
                    EVENTS REGISTRY

                    E. Antonucci1, O. Paoletti2, W. Ageno3, M. Righini4,
                                                                                       us
                                                                                   administered in 10 patients (6 ICH and 4 GIB); andexanet
                                                                                   alpha in 2 patients with ICH. Creatinine, blood cell count,
                                                                                   and PT/aPTT were the most frequent tests requested
                                                                                   al
                    P. Verhamme5, G. Colombo6, P. Sivera7, M.S. Cotelli8,          (85%), while specific DOAC measurements were per-
                    M. Tala2, R. Morandini2, C. Dalla Noce2, D. Poli9,             formed only in 30% of patients. At hospital discharge,
                                                                           ci

                    G. Palareti1, S. Testa2                                        63% of all patients had complete resolution of bleeding
                                                                                   signs.When we consider ICH patients only, we observed
                                                                     er


                    1Fondazione Arianna Anticoagulazione,      Bologna, Italy;     complete resolution of symptoms in about 58% of patients
                    2Dipartimento    di Medicina di Laboratorio, Centro            and residual disability in 46/111patients (41%). During
                                                               m



                    Emostasi e trombosi, ASST Cremona, Cremona, Italy;             hospitalization the mortality rate was 11%. At 6 months,
                    3Dipartimento di Medicina e Chirurgia, Università degli        the complete resolution was recorded in 78% of patients.
                                                  om




                    Studi dell’Insubria, Varese, Italy; 4Division of Angiology     Overall mortality rate of the study was 14.3%.
                    and Hemostasis, Department of Medical Specialties,             Conclusions: Our data show that the therapeutic strategies
                    Geneva University Hospital and Faculty of Medicine,            for management of bleeding are mainly symptomatic
                                               -c




                    Geneva, Switzerland; 5Center for Molecular and                 treatment of blood loss, treatment of the anatomical cause
                    Vascular Biology, Department of Cardiovascular                 of bleeding and the use of specific reversal agent as sug-
                                       on




                    Sciences, University of Leuven, Leuven, Belgium; 6DB           gested by current guidelines. In a very low number of
                    Internistica e Centro Trombosi ed Emostasi, AO di              patient we registered a conservative approach. Nearly
                    Circolo, ASST dei Sette Laghi, Varese, Italy; 7SCDU            one-quarter of patients received an inappropriate dose of
                                  N




                    Ematologia E Terapie Cellulari Ospedale Mauriziano             DOAC. A low rate of specifc drug measurements is still
                    Umberto I di Torino, Torino, Italy; 8 SSD Neurologia           reported, despite the current guidelines for management of
                    ASST Valcamonica Esine, Brescia, Italy; 9 SOD Malattie         DOAC associated bleeding are being updated to perform
                    Aterotrombotiche, Azienda Ospedaliero Universitaria-           the measurement in this specifc clinical conditions.
                    Careggi, Firenze, Italy

                    Background and Aims: The management of major                   OC091
                    bleeding in patients treated with direct oral anticoagulants
                    (DOACs) is a serious medical concern. START-Events,
                    prospective, observational, multicenter, international         GENDER DIFFERENCE AND RISK OF MAJOR
                    studyand a branch of the START registry (Survey on             BLEEDING IN PATIENTS ON ANTI-THROMBOTIC
                    anTicoagulatedpAtientsRegisTer) (NCT02219984), aims            TREATMENT FOR MECHANICAL HEART VALVE
                    to describe the management of bleeding or recurrent            PROSTHESIS. ANALYSIS CONDUCTED AT THE
                    thrombotic events in routine clinical practice. We here        SALAM CENTRE FOR CARDIAC SURGERY -
                    present the results of the management of bleeding patients.    EMERGENCY NGO – KHARTOUM, SUDAN
                    Methods: Clinical characteristics of patients, laboratory
                    data at admission and during follow-up, site of bleeding,      N. Erba1,2,3, A. Mocini2, A. Sawi2, F. Hamuda2,
                    therapeutic strategies and outcomes at the time of hospital    M. Langer1, G. Portella1, F. Masini2, S. Testa3, D. Poli3




                                                                [Hematology Reports 2021; 13:s3]                                                  [page 63]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 64



         Oral communications




                     1Emergency   NGO ONLUS, Milan, Italy; 2SALAM                OC092
                     Centre for Cardiac Surgery, Khartoum, Sudan;
                     3Federazione Centri per la Diagnosi Della Trombosi e

                     la Sorveglianza delle Terapie Antitrombotiche (FCSA),       PREDICTIVE ABILITY OF THE HAS-BLED SCORE
                     Milan, Italy                                                OF THE NEWLY PUBLISHED SWISS-AF SCORE IN
                                                                                 ATRIAL FIBRILLATION PATIENTS ON
                     Background and Aims: Anticoagulant therapy with             ANTICOAGULANT TREATMENT WITH VITAMIN K
                     Warfarin exposes the patient to the risk of major bleed-    ANTAGONITS AND DIRECT ORAL
                     ing, and consequently, to life-threatening risks, tempo-    ANTICOAGULANTS. PRELIMINARY RESULTS OF
                     rary or permanent disabilities and thrombotic risk relat-   THE START2 AF REGISTER
                     ed to temporary withdrawal of anticoagulant therapy.
                     The study was aimed to analyze gender-related risk of       D. Poli1, L. Sechi2, E. Antonucci3, F. Crudele2,
                     major bleeding and to investigate the gender-related        M. Berteotti2,G. Colombo4, L. Masciocco5, T. Lerede6,
                     differences in patients on Warfarin for mechanical heart    A. Toma7, P. Pignatelli8, D. Pastori8, S. Testa9,
                     valve (MHV) prostheses operated at the Salam Centre         R. Marcucci2, G. Palareti3
                     for Cardiac surgery in Khartoum. Methods: Between
                     August 1, 2018 and September 30, 2019 all patients fol-     1SOD  Malattie Aterotrombotiche, Azienda Ospedaliero
                     lowed by the Anticoagulation Clinic of the Salam            Universitaria-Careggi, Firenze; 2Dipartimento di
                     Centre for Cardiac surgery were included in a prospec-      Medicina Sperimentale e Clinica, Università degli Studi




                                                                                                         ly
                     tive observational study to collect data on the adherence   di Firenze; 3Fondazione Arianna Anticoagulazione,
                     and quality of the anticoagulant treatment. Bleeding        Bologna; 4DB Internistica e Centro Trombosi ed




                                                                                                  on
                     events occurred during follow-up were recorded, Major       Emostasi, AO di Circolo, ASST dei Sette Laghi, Varese;
                     Bleedings were defined according to ISTH criteria.          5UO Medicina Interna, Presidio Ospedaliero Francesco
                     Patients with less than three months of observation         Lastaria di Lucera, ASL Foggia, Lucera (FG); 6USC




                                                                                            e
                     were excluded from the observation. Patients with inter-    SIMT, Centro Emostasi e Trombosi, Ospedale Papa
                     vals between controls longer than 3 months were
                     defined “incompliant”. Patients who omit the check
                     INR for 6 months or more, despite follow-up calls, were
                                                                                     us
                                                                                 Giovanni XXIII, Bergamo; 7UO Laboratorio Analisi
                                                                                 Distretto Ovest, ULSS 8 Berica, Arzignano (VI);
                                                                                 8Department of Clinical, Internal, Anesthesiological and
                                                                                 al
                     classified “lost” at follow-up. Results: We studied 3647    Cardiovascular Sciences, Sapienza University of Rome,
                     patients (median age 25.7 years; females 53.9%) who         Roma; 9Dipartimento di Medicina di Laboratorio,
                                                                         ci

                     were on anticoagulant treatment with warfarin after         Centro Emostasi e trombosi, ASST Cremona, Cremona,
                     MHV implantation. Patients aged <14 y were 19% of           Italy
                                                                   er


                     the included males and 16.1% of the included females
                     (p=0.02). Aortic valves were implanted in 23.8% of          Background and Aims: Oral anticoagulant therapy
                                                              m



                     males and 7.2% of females (p value <0.001); mitral          (OAT) is recommended in patients with atrial fibrillation
                     valves were implanted in 40.5% of males and 68.4% of        (AF) due to the associated risk of ischemic stroke.
                                                  om




                     females (p value <0.001); mitral-aortic valves were         However, the stratification of the bleeding risk associated
                     implanted in 35.7% of males and 24.4% of females (p         with OAT is still not optimal. The HAS-BLED is the
                     value <0.001). 80% of patients received also aspirin (75    most widely used bleeding score, but it demonstrate a
                                               -c




                     or 100 mg OD) with not significant differences between      low predictive ability. The SWISS-AF score to predict
                     males and females (respectively 79.4 vs 81.2, p=0.2).       bleeding risk has been published. The score is easy to cal-
                                       on




                     Females showed worse Time in Therapeutic Range              culate and includes 4 items (age≥75 years, hypertension,
                     (median F 50% vs M 56%, p value <0.001), instead they       previuos major bledings, cancer).The purpose of the
                     had better compliance to therapy (uncompliant F 6.8%        study is to compare the predictive ability HAS-BLED
                                   N




                     vs M 10.2%, p value <0.001) and less of them were lost      and of the SWISS-AF score in AF patients enrolled in the
                     at follow-up (F 2.0 vs M 3.4, p value=0.01). During the     observational, prospective START2-Register – Survey
                     observational period we recorded 85 major bleeding          on anTicoagulated pAtients RegisTer. Methods: We
                     episodes (2.16 % p-y, n. 9 fatal): 16 were cerebral, 19     investigated the clinical characteristics of AF patients
                     were gastrointestinal, and 32 were menometrorrhagia.        receiving included in the Register and we apply the dif-
                     At univariate analysis, female gender and aspirin treat-    ferent scores. Thrombotic events, major and clinically
                     ment were significantly associated with bleeding risk       significant relevant not major bleeding events and death
                     (OR 2.2, 95%CI 1.4-3.4; p value <0.001 and OR 1.8,          occurred in the follow-up were recorded. Major, clinical-
                     95%CI 1.0-3.4; p value=0.05, respectively). When            ly relevant non-major bleedings and thrombotic events
                     gynaecological bleedings were excluded, women were          were recorded. Predictive ability for bleeding was calcu-
                     not at a higher risk of bleeding respect to men (OR 1.2,    lated for both scores among patients on VKAs and
                     95%CI 0.7-1.9; p value=0.5). Conclusions: Women of          patients on DOACs. Results: We studied 6165 patients
                     childbearing age on anticoagulant therapy are particu-      (54% males), median age 75.6 years, 34.1% of whom
                     larly exposed to the risk of major predominantly gynae-     were on direct oral anticoagulants (DOACs). During fol-
                     cological bleedings. The use of associated aspirin treat-   low-up 179 major bleedings were recorded (rate 1.46
                     ment should be carefully evaluated, especially in           x100 patient years; rate 1.28 and 1.89 for patients on
                     women. Specific gynaecological programs should be           VKAs and on DOACs respectively. At multivariate
                     implemented for anticoagulated women.                       analysis the history of previous bleeding, heart failure,




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                                                                            XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    the history of previous stroke/TIA, the concomitant use       OC094
                    of antiplatelet therapy, the presence of active cancer were
                    significantly associated with bleedings. The predictive
                    ability calculated with the c statistic (95% CI) for major    MANAGEMENT OF PATIENTS WITH THROMBOTIC
                    haemorrhage for HASBLED was: 0.56 (0.51-0.62),                EVENTS DURING TREATMENT WITH DIRECT ORAL
                    p=0.03 when patients on VKA were considered; 0.59             ANTICOAGULANTS
                    (0.52-0.66), p=0.01 when patients on DOAC were con-
                    sidered. For the Swiss-AF score the c statistic (95% CI)      G. Cammà¹, E. Antonucci3, D. Poli4, M. Tala5,
                    was: 0.55 (0.5-0.6), p=0.03 when VKAs were considered         O. Paoletti5, G. Palareti3, S. Testa5, W. Ageno2
                    and 0.60 (0.52-0.67), p=0.007 when DOACs patients             1Department
                    were considered. Conclusions: The main result of the                         of Internal Medicine, Fondazione
                    study is that the predictive ability for bleeding of SWISS-   Policlinico Universitario AGemelli IRCCS, Università
                    AF score and of HAS-BLED score are similarly modest,          Cattolica del Sacro Cuore, Rome, Italy; 2Department
                    both either in VKAs patients or in DOACs patients.            of Emergency Medicine, Fondazione Macchi,
                                                                                  Università dell’Insubria, Varese, Italy; 3Fondazione
                                                                                  Arianna, Bologna, Italy; 4Azienda Ospedaliera
                    OC093                                                         Careggi, Florence, Italy; 5Azienda Ospedaliera di
                                                                                  Cremona, Cremona, Italy




                                                                                                          ly
                    INTRACRANIAL HEMORRHAGE IN PATIENTS                           Background and Aims: Direct oral anticoagulants
                    WITH PRIMARY OR METASTATIC BRAIN CANCER                       (DOACs) are effective in the treatment of venous




                                                                                                  on
                    TREATED OR NOT WITH ANTICOAGULANTS: A                         thromboembolism (VTE) and in the prevention of car-
                    SYSTEMATIC REVIEW AND META-ANALYSIS                           dioembolic events in patients with atrial fibrillation
                                                                                  (AF). Little evidence exists on the management of




                                                                                             e
                    M. Giustozzi1, G. Proietti2, C. Becattini, F. Roila3,         patients who present with acute thrombotic events dur-
                    G. Agnelli, M. Mandalà3
                    1Internal, Vascular and Emergency Medicine - Stroke
                                                                                      us
                                                                                  ing treatment with DOACs. Methods: We conducted a
                                                                                  prospective observational study on patients started on
                                                                                  anticoagulant treatment at Italian Thrombosis Clinics
                    Unit, University of Perugia, Perugia, Italy; 2Medical         (START-Register). Patients diagnosed with an arterial
                                                                                  al
                    student, University of Perugia, Perugia, Italy; 3Unit of      or venous thrombotic event during treatment with
                                                                            ci

                    Medical Oncology, University of Perugia, Perugia, Italy       DOACs were identified. Information on management
                                                                                  strategies and outcomes was collected. Results: We
                                                                    er


                    Introduction: Anticoagulant treatment in patients with        enrolled 110 patients with a median age of 71 years, 65
                    primary and metastatic brain cancer is a concern due to       (60%) were males, 70 (64%) had AF, 40 (36%) had
                                                              m



                    risk of intracranial hemorrhage (ICH). We performed a         VTE. Of these patients, 53 were receiving low dose
                    systematic review and meta-analysis to evaluate the risk      DOACs, only 19 according to the label. Median time
                                                  om




                    of ICH in patients with primary or metastatic brain can-      from starting DOAC to thrombotic events was 230 days
                    cer treated or not with anticoagulants. Methods:              (IQR 72-585); 266 days (IQR 122-710) in AF group
                    Articles on ICH in patients with primary or metastatic        and 117 days (IQR 35-389) in the VTE group. In the AF
                                               -c




                    brain cancer treated or not with anticoagulants pub-          group the majority of patients had ischemic stroke
                    lished up to September 2021 were identified by search-        (n:46), in the VTE group recurrent VTE was the most
                                       on




                    ing PUBMED, EMBASE and CENTRAL.The primary                    common event (n:32). Overall, there were 64 arterial
                    outcome of this analysis was ICH. Results: Thirty stud-       thrombotic events: 49 strokes, 6 transient ischemic
                    ies were included. Rate of ICH was 13.0% in 1,009             attacks (TIA), 6 peripheral emboli, and 3 acute myocar-
                                  N




                    patients with metastatic and 6.4% in 2,353 patients with      dial infarctions (AMI). DOACs therapy was stopped in
                    primary brain cancer [Relative risk (RR) 3.26, 95% CI         40 patients (34 strokes, 5 TIA and 1 AMI). Low molec-
                    2.69-3.94; I2 92.8%). In patients with primary brain          ular weight heparin (LMWH) was started in 17 strokes,
                    cancer, ICH occurred in 12.5% and 4.4% of patients            1 peripheral embolism and 1 AMI. LMWH in addition
                    treated or not treated with anticoagulants, respectively      to aspirin was used in 14 strokes and 1 TIA; aspirin
                    [11 studies, 659 treated and 1,346 not treated patients,      alone in 8 strokes and 2 TIA. Nine patients (5 stroke
                    RR 2.63, 95% CI 1.48-4.67, I2 49.6%]. In patients with        and 4 peripheral emboli) received thrombolysis or
                    metastatic brain cancer, ICH occurred in 14.7% and            thrombectomy. At discharge, 18 patients (10 strokes, 4
                    15.4%, respectively (5 studies, 265 treated and 301 not       TIA, 2 peripheral embolism and 2 AMI) continued with
                    treated patients, RR 0.92, 95% CI 0.43-1.93, I2 0%).          the same anticoagulant regimen used before the event;
                    ICH occurred in 8.3% of 172 treated with direct oral          7 patients (6 strokes and 1 TIA) added aspirin to the
                    anticoagulant (DOAC) and in 11.7% of 278 treated with         DOAC. DOAC dose was increased, while maintaining
                    low-molecular weight heparin (LMWH) (5 studies, RR            the same molecule, in 6 strokes and 1 TIA. A different
                    0.44, 95% CI 0.25-0.79, I2 0%). Conclusions: Patients         DOAC was prescribed to 15 patients (13 strokes and 2
                    with metastatic brain cancer have a particularly high         peripheral embolism). A different DOAC plus aspirin to
                    risk of ICH. Patients with primary brain cancer have an       4 strokes and 1 peripheral embolism. Vitamin K antag-
                    increased risk of ICH during anticoagulation. DOACs           onists (VKAs) were given to 8 patients (6 strokes, 1
                    are associated with a lower risk of ICH than LMWH.            peripheral embolism and 1 AMI). Four patients died, all




                                                               [Hematology Reports 2021; 13:s3]                                             [page 65]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 66



         Oral communications




                     after stroke, 24 stroke patients had residual disability.     no certain evidence regarding the peri-operative manage-
                     Among the 46 venous thrombotic events, there were 21          ment of elderly patients taking OAC- vitamin K antago-
                     deep vein thrombosis (DVT), 9 DVT plus pulmonary              nists (VKAs) or direct oral anticoagulants (DOACs) and
                     embolism (PE),11 isolated PE, 4 superficial vein throm-       antiplatelets- that require major orthopedic surgery. To
                     bosis (SVT) and 1 retinal vein occlusion. All patients        date, there is no consensus on the timing and period of
                     stopped DOACs therapy. LMWH therapy was started in            OAC/antièlatelets interruption and re-initiation of thera-
                     41 cases (20 DVT, 6 DVT, 11 isolated PE, 4 SVT) and           py in these patients. Aims of the ORTHO-START reg-
                     fondaparinux in 3 (1 DVT and 2 DVT+PE). At dis-               istry are: 1) To collect data on the peri-operative manage-
                     charge, LMWH was continued in 7 patients (2 DVT, 3            ment of anticoagulation therapies in elderly patients who
                     DVT+PE, 1 PE, 1 SVT). The same DOAC was con-                  need emergency or elective orthopedic surgery; 2)To
                     firmed in 19 patients (11 DVT, 2 DVT+PE, 6 PE); the           investigate transfusion practices in the context of emer-
                     dose was increased in two patients. A different DOAC          gency or elective orthopedic surgery in elderly patients,
                     was prescribed to 5 DVT, 1 DVT+PE and 2 PE; 9                 with the objective to collect information useful to set up
                     patients were switched to VKAs (2 DVT, 3 DVT+PE, 1            a patient blood management program. Methods: The
                     PE, 2 SVT). No patient died.                                  ORTHO-START is a multicentre, prospective, observa-
                     DOACs activity at baseline was measured in 35                 tional study investigating management of patients taking
                     patients. It was less than 50 ng / ml in 12 patients: 8       OAC and / or antiplatelet agents requiring elective inter-
                     strokes and 4 VTE; less than 30 ng / mL in 3 patients, 2      vention of hip replacement or knee or urgent surgery for
                     were receiving low dose DOACs inappropriately.                hip fracture. Data collection is web-based. Patients not




                                                                                                           ly
                     Conclusions: The management of thrombotic events              taking OAC or antiplatelets and undergoing the above-
                     during DOACs therapy is heterogeneous and varies              listed procedures have been simultaneously recruited




                                                                                                    on
                     between arterial and venous events. A substantial pro-        with the objective to compare clinical outcomes and lab-
                     portion of patients were discharged with the same drug        oratory characteristics to those of patients taking OAC
                     and dosage. DOACs activity at the time of the event           and/or antiplatelets. Six Centres have recruited patients (




                                                                                              e
                     was measured in only one third of patients.                   July 2018- March 2022). Results: Of 1050 patients


                     OC095
                                                                                       us
                                                                                   recruited�(median age 81, IQR 73-87; BMI: 25.7, range
                                                                                   15-46) ], 755 (72%) were women. 635 (60.5%) had
                                                                                   chronic hypertension, 194 diabetes (171 of them suffered
                                                                                   from hypertension and diabetes), 266 (25.3%) suffered
                                                                                   al
                                                                                   from cardiovascular disease. At admission, hemoglobin
                     FINDINGS FROM THE ORTHO-START REGISTRY:
                                                                           ci

                                                                                   (Hb) median values were 12.2 g/dl (IQR 5-17), 794
                     PERIOPERATIVE MANAGEMENT OF                                   (80.1%) patients showed values <10.5 g/dl. Non-0 group
                                                                     er


                     ANTICOAGULANT THERAPIES IN ELDERLY                            was recorded in 641 (65.3%) patients. Regarding surgical
                     PATIENTS UNDERGOING MAJOR ORTHOPEDIC                          procedure, we have completed information on 871
                                                               m



                     SURGERY                                                       patients, 356 (40.9%) of whom underwent an urgent sur-
                                                                                   gery. Furthermore, 326 (37.4%) patients were taking
                                                   om




                     E. Grandone, E. Antonucci, D. Colaizzo,                       antiplatelets and 247 ( 28.4%) OAC (185 of them for atri-
                     A. De Laurenzo, C. Rostagno, F. Marongiu,                     al fibrillation). The attached Table 1 summarizes use of
                     F. Gorgoglione, G. Maccagnano, G. Di Rienzo,                  peri-operative thrombo-prophylaxis, transfusion rate and
                                                                                   follow-up on the entire cohort.
                                                -c




                     L. Mirabella, M. Senzolo, A. Ostuni, G. Palareti
                                                                                   Table 1.
                                        on




                     Fondazione IRCCS Casa Sollievo della Sofferenza, San
                     Giovanni Rotondo, Italy; The First IM Sechenov
                     Moscow State Medical University, Obstetrics &
                                   N




                     Gynecology, Moscow, Russian Federation; University of
                     Foggia; Fondazione Arianna, Bologna, Italy; University
                     of Florence, Italy; Hospital F. Perinei, Bari, Italy;
                     University of Cagliari, Italy; Padua University Hospital,
                     Italy; A.O.U. Policlinico di Bari, Transfusion Medicine,
                     Bari, Italy

                     Background: Peri-operative management of oral antico-
                     agulant therapy in the context of emergency orthopedic
                     surgery for femur fracture or elective major orthopedic
                     surgery for hip/knee replacement in elderly patients, is      Mortality rate was not significantly higher in those tak-
                     challenging, because of the presence of multiple co-mor-      ing vs “not taking” OAC/antiplatelets ( 1.0 vs 1.3),
                     bidities and medical treatments. For this reason in elderly   whereas overall complications were significantly high-
                     patients, interruption and re-initiation of the Oral          er in patients taking OAC/antiplatelets (p<0.001).
                     Anticoagulant (OAC) and antiplatelets therapy require a       Transfusion rate was not dependent on chronic OAC or
                     multidisciplinary team. In the context of peri-operative      antiplatelets therapy, as among the 339 transfused, only
                     clinical management, OAC therapy raises the issue of          93 (27.4%) were taking OAC/antiplatelets.
                     bleeding risk, which is higher in these patients. There is    Conclusions: This registry gives a snapshot on peri-




         [page 66]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 67



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    operative management of Italian patients undergoing         bid patients. General supportive measures seem to be
                    major orthopedic surgery. This preliminary analysis         effective in the most of cases. The rate of death was
                    shows that mortality rate, number of complications and      clinically relevant.
                    transfusion rate in patients taking OAC or antiplatelets
                    are not higher that those observed in other patients.
                                                                                OC097

                    OC096                                                       PROSPECTIVE EVALUATION OF CANCER
                                                                                PATIENTS AT HIGH THROMBOTIC RISK DURING
                    MANAGEMENT AND OUTCOMES OF PATIENTS                         ANTICANCER TREATMENT WITH KHORANA AND
                    WITH DOAC-RELATED GASTROINTESTINAL                          VIENNA-CATS NOMOGRAM SCORES
                    BLEEDING: FINDINGS FROM PROSPECTIVE
                    COHORT STUDY                                                C. Verzeroli1, M. Marchetti1, C. Giaccherini1, L. Russo1,
                                                                                S. Gamba1, C.J. Tartari1, S. Bolognini1, C. Ticozzi1,
                    M. Verso1, O. Morelli2, D. Castellani2, G. Bassotti2,       A. Santoro2, F. De Braud3, G. Gasparini4, F. Petrelli5,
                    A. Vinci1, C. Becattini1                                    F. Giuliani6, A. D’Alessio7, M. Minelli8, R. Labianca9,
                                                                                C. Morlotti10, P. Malighetti10, D. Spinelli11, A. Falanga1,12
                    1Emergency     and Vascular Medicine-Stroke Unit,           on behalf of the HYPERCAN Investigators*




                                                                                                         ly
                    University of Perugia; 2Gastroenterology Division,
                    University of Perugia, Italy                                1Immunohematology      and Transfusion Medicine, ASST




                                                                                                 on
                                                                                Papa Giovanni XXIII, Bergamo, Italy; 2Oncology Unit,
                    Background and Aims: Direct oral anticoagulants             IRCCS Humanitas Institute, Rozzano, Italy; 3Oncology
                    (DOACs) have been associated with an increased risk         Unit, IRCCS National Cancer Institute, Milan, Italy;




                                                                                           e
                    of gastrointestinal bleeding (GIB). The aim of the study    4Oncology Unit, Hospital San Filippo Neri, Rome, Italy
                                                                                5Oncology
                    was to prospectively collect data on real-life manage-
                    ment and outcomes of DOACs-related GIB. Methods:
                    A prospective, cohort study with a 6-month follow-up
                                                                                    us         Unit, Hospital Treviglio-Caravaggio,
                                                                                Treviglio, Italy; 6Oncology Unit, IRCCS Cancer Institute
                                                                                Giovanni Paolo II, Bari, Italy; 7Medical Oncology and
                    (FUP) was performed. Results: From November 2013            Internal Medicine, Policlinico San Marco, Zingonia-
                                                                                al
                    to March 2022, 208 patients (121 male) with DOACs-          Bergamo, Italy; 8Oncology Unit, Hospital San Giovanni
                                                                                Addolorata, Rome, Italy; 9DIPO, ASST Papa Giovanni
                                                                        ci

                    related GIB were included. At entry, the mean patient
                    age was 83±4 years. 63 (30.1%%) patients presented          XXIII, Bergamo, Italy; 10Università degli Studi di
                                                                  er


                    GIB on dabigatran (low dose in 45), 73 (35.1%) on           Bergamo, Bergamo, Italy; 11Università degli Studi
                    rivaroxaban (low dose in 35) 48 (23.1%) on apixaban         Milano-Bicocca, Milan, Italy; 12University of Milan
                                                             m



                    (low dose in 15) and 24 (11.5%) on edoxaban (low dose       Bicocca, School of Medicine, Italy
                    in10). 9.6% received inappropriate doses of DOACs.
                                                 om




                    The indication for DOAC treatment was atrial fibrilla-      Background and Aims: Identifying cancer outpatients
                    tion (AF) in 87.5% of patients, venous thromboem-           at high risk of venous thromboembolism (VTE), in
                    bolism in 9.1% and both indication in 3.4% of patients.     whom anticoagulant thromboprophylaxis is justified, is
                                              -c




                    In AF patients, median CHA2DS2-VASC was 4.5 and             an unmet clinical need. The use of validated risk assess-
                    median HAS-BLED was 2.6. The mean number of con-            ment models (RAMs) involving clinical and biological
                                      on




                    comitant drugs was 6. According to the ISTH criteria,       parameters can be a relevant approach to hit this goal.
                    GIB was classified as major in 152 patients and as clin-    Among several proposed RAMs, the Khorana risk score
                    ically relevant non-major bleeding in 56 patients. The      (KRS) and the Vienna-CATS nomogram score have been
                                  N




                    bleeding site was in upper GI tract for 79 patients         developed and externally validated in newly diagnosed
                    (42.5%), in lower gastrointestinal tract in 65 patients     ambulatory cancer patients and currently appear to be the
                    (34.9%), in both upper and lower tract in 42 patients       most promising ones. In a large prospective cohort of
                    (22.6%). In 22 patients (10.6%), the endoscopic proce-      1,286 outpatients with metastatic non-small cell lung,
                    dures were not performed or were not able to detect the     colorectal, gastric, or breast cancers undergoing
                    source of bleeding. All patients discontinued the DOAC      chemotherapy [the HYPERCAN study], we aimed to
                    administration, except one. 143 patients were trans-        describe the incidences of objectively verified VTE and
                    fused with packed red cells, with a median number of        compare the discriminatory performance of the two vali-
                    blood transfusions of 2.4. A reversal strategy was adopt-   dated RAMs (KRS and the Vienna-CATS nomogram) for
                    ed in 13.5% of patients. The endoscopic procedures          the identification of high VTE risk subjects. Methods:
                    were performed within 24 hours after admission in           KRS and Vienna-CATS nomogram score were applied to
                    52.0% of cases. Fifty-four patients received an endo-       the HYPERCAN cohort using pre-chemotherapy patient
                    scopic haemostasis. The anticoagulant treatment was         variables (leucocyte and platelet counts, hemoglobin
                    discontinued in 50 patients. During the 6-month FUP,        level, and BMI for KRS and D-dimer levels for Vienna-
                    12 patients had a bleeding recurrence and 8 patients had    CATS nomogram). Only VTE events within 6 months,
                    thrombotic complications. Six patients died during hos-     confirmed and validated by the Independent Central
                    pitalization and 45 during 6-month FUP. Conclusions:        Adjudication Committee, were included in the analysis.
                    Patients with DOAC-related GIBs were old and comor-         The actual VTE rate according to KRS and Vienne-




                                                             [Hematology Reports 2021; 13:s3]                                                   [page 67]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 68



         Oral communications




                     CATS nomogram score was estimated by the Kaplan-              Recently, Pennisi and colleagues reported that a modified
                     Meier method considering death as a competing event.          version of EASIX (mEASIX)- combining platelets, CRP,
                     Results: One hundred-twenty objectively confirmed             and LDH - correlated with CRS and oncological out-
                     VTE (10.3%) were recorded, including isolated deep            comes. Firstly, we measured a baseline coagulative asset
                     vein thrombosis (DVT, 45.8%), pulmonary embolism              on day 0 (D0) before CAR-T reinfusion, and we com-
                     (PE, 42.5%), and PE+DVT (11.7%). By ROC analysis,             pared it with a coagulative pattern sampled on day 4
                     KRS provided a non-relevant AUC of 0.39 for 6-months          (D4). We then calculated mEASIX for every other day
                     VTE. Cumulative VTE incidence was 6.0 % (95% CI               following CAR-T cells reinfusion, we graded daily CRS
                     3.8-9.7) for KRS<2 and 12.3% (95% CI 9.9-15.3) for            and ICANS and we analyzed correspondent coagulation
                     KRS≥2 (p=ns). Differently, Vienna-CATS nomogram               parameters. Results: Between D0 and D4, we found a
                     score provided an AUC of 0.63 for 6-months VTE.               significant decrease in antithrombin and platelets, and a
                     Cumulative VTE incidence was 7.2% (95% CI 5.6-9.2)            concurrent increase of aPTT, INR, and D-dimer (paired
                     and 17% (95% CI 13.5-21.3) in low- and high-risk              t-test, p<0.001, <0.001, 0.001, <0.001, and 0.01 respec-
                     patients, respectively (SHR=2.4, p<0.001). Conclusions:       tively) (Figure 1A). Fibrinogen did not significantly
                     In the prospective cohort of metastatic cancer patients of    modify between D0 and D4 (median 390 vs 427 mg/Dl,
                     the HYPERCAN study, the KRS showed a low perform-             p=0.36); nevertheless, fibrinogen is highly specific in
                     ance. Differently, the Vienna-CATS nomogram score sig-        predicting consumptive coagulopathy but shows low
                     nificantly categorized patients at low VTE risk, in which     sensitivity in highly inflammatory conditions. We
                     the increased risk of bleeding due to thromboprophylaxis      hypothesized that fibrinogen remained stable due to




                                                                                                           ly
                     would outweigh benefits, and high VTE risk, which             opposite trends originating from activated consumptive
                                                                                   coagulopathy and acute inflammatory state. To corrobo-




                                                                                                    on
                     might benefit from thromboprophylaxis. Therefore, the
                     inclusion of D-dimer in RAM, as in the Vienna-CATS            rate this hypothesis, we verified that fibrinogen and CRP
                     nomogram, seems to be a promising approach toward             were positively correlated even in the post-infusional set-
                     reliable and clinically applicable RAMs for VTE predic-       ting of CAR-T cells treatment (p<0.001). On D4, 5/20




                                                                                               e
                     tion in these widespread cancers.                             (25%) evaluable patients presented an ISTH score of 5,
                     Funded by Italian Association for Cancer Research
                     (AIRC) “5xMILLE” n.12237 grant.                                   us
                                                                                   consistent with disseminated intravascular coagulation
                                                                                   (DIC). This score takes into account platelet count, fib-
                                                                                   rinogen, PT, and D-dimer; however, low platelet count
                                                                                   al
                                                                                   may also be due to the previous fludarabine-based lym-
                     OC098                                                         phodepletion in this setting, and DIC scoring systems
                                                                           ci

                                                                                   may not be performant. Greater mEASIX correlated with
                                                                                   higher aPTT, D-dimer, fibrinogen, and von Willebrand
                                                                     er


                     CONSUMPTIVE COAGULOPATHY AFTER                                factor, and with lower antithrombin (Figure 1B). CRS
                     TREATMENT WITH CAR-T CELLS CORRELATES                         graded more than 1 was associated with higher aPTT, fib-
                                                               m



                     WITH MARKERS OF ENDOTHELIAL ACTIVATION                        rinogen, factor VIII and von Willebrand factor.
                                                                                   Occurrence of neurotoxicity (ICANS) correlated with
                                                   om




                     E. Galli1,2, F. Sorà1,2, F. Autore2, E. Metafuni2,            higher D-dimer and showed a trend with higher aPTT. In
                     I. Innocenti2, A. Fresa1, M.A. Limongiello2,                  our cohort, no patients experienced major bleedings, and
                     S. Giammarco2, L. Laurenti1,2, A. Bacigalupo1,2,              in one case, we observed a catheter-related deep vein
                     P. Chiusolo1,2, V. De Stefano1,2, S. Sica1,2
                                                -c




                                                                                   thrombosis (DVT) with pulmonary embolism (PE).
                     1Sezione
                                        on




                               di Ematologia, Dipartimento di Scienze
                     Radiologiche ed Ematologiche, Università Cattolica del
                     Sacro Cuore, Roma; 2Area Ematologica, Dipartimento
                                   N




                     di Diagnostica per Immagini, Radioterapia Oncologica
                     ed Ematologia, Fondazione Policlinico Universitario A.
                     Gemelli IRCCS, Roma, Italy

                     Background and Aims: Cytokine release syndrome
                     (CRS) and classical consumptive coagulopathy are asso-
                     ciated in up to 56% of patients treated with chimeric anti-
                     gen T (CAR) cells. In this report, we describe coagulative
                     patterns and laboratory parameters of endothelial activa-
                     tion following CAR-T cells reinfusion. Methods: Here
                     we report 27 consecutive patients with aggressive lym-        Figure 1.
                     phoproliferative diseases treated with CAR-T cells in our
                     center from September 2019 to April 2022. The main            Conclusions: Overall, we observed a laboratory con-
                     diagnoses were diffuse large B cell lymphoma and pri-         sumptive coagulopathy immediately after CAR-T cells
                     mary mediastinal B cell lymphoma. In the setting of           reinfusion, lacking a fibrinogen decrease possibly due
                     hematopoietic stem cell transplantation, endothelial acti-    to the high inflammatory burden. This altered coagula-
                     vation has been correlated to an easy bedside score, the      tive asset figures as a laboratory – and not clinical- con-
                     Endothelial Activation and Stress Index (EASIX).              dition. Endothelial hyperactivation may play a key role,




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 69



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    enforced by the close connection between mEASIX,            273 pts. ET in 406, pre-MF in 53, MF in 57, and unclas-
                    coagulopathy, and the increase of von Willebrand fac-       sifiable MPN in 4; 710 and 83 pts received ASA and
                    tor. The endothelial hyperactivation needs to be framed     TKL/CPG as unique antithrombotic treatments during
                    in the scenario of sustained moderate-to-severe CRS.        4406 and 531 observation years, respectively.
                                                                                Thromboses were 84 (arterial/venous 20/64) in the ASA
                                                                                group and 19 (A/V 8/11) in the TKL/CPG group. The IR
                    OC099                                                       of thrombosis per 100 pt-years was 1.91 and 3.58 under
                                                                                ASA and TKL/CPG, respectively (p=0.01). The hazard
                                                                                ratio for thrombosis in the TKL/CPG versus ASA pts
                    EFFECTIVENESS AND SAFETY OF THE
                                                                                was 2.36 (95%CI 1.25-4.45, p=0.007) (Figure 1). There
                    ANTI-P2Y12 TICLOPIDINE AND CLOPIDOGREL
                                                                                was a significant difference between ASA and
                    AGENTS IN PHILADELPHIA-NEGATIVE                             TKL/CPG in the IR per 100 pt-years of arterial throm-
                    MYELOPROLIFERATIVE NEOPLASMS IN                             bosis (0.45 vs. 1.51, p=0.002), but not for venous throm-
                    COMPARISON WITH ASPIRIN                                     boses (1.45 vs. 2.1, p=0.3). In ET pts, the IR per 100 pt-
                                                                                years of thrombosis was 1.48 and 3.7 under ASA and
                    F. Ramundo, S. Ceglie, S. Betti, T. Za, A. Ciminello,       TKL/CPG, respectively (p=0.02). In contrast, we detect-
                    F. Bartolomei, F. Di Landro, P. Arcuri, P. Chiusolo,        ed no significant difference in the PV and pre-MF/MF
                    E. Rossi, V. De Stefano                                     cohorts (p=0.3 and p=0.8). We recorded 63 major bleed-
                                                                                ings in the ASA group and 3 in the TKL/CPG group (IR




                                                                                                        ly
                    Sezione di Ematologia, Dipartimento di Scienze              per 100 pt-years 1.43 vs. 0.56, p=0.1). Conclusions: In
                    Radiologiche ed Ematologiche, Università Cattolica,




                                                                                                on
                                                                                MPN pts, the anti-P2Y12 antiplatelet agents TKL and
                    Fondazione Policlinico AGemelli IRCCS, Roma, Italy          CPG are less effective than ASA in preventing thrombo-
                                                                                sis; this finding is more evident for arterial thrombosis
                    Background and Aims: Aspirin (ASA) is widely                and ET patients. No difference was found in the IR of




                                                                                            e
                    employed in patients with Philadelphia-negative myelo-      major bleeding events. Therefore, special attention
                    proliferative neoplasms (MPN) for antithrombotic pro-
                    phylaxis. Its efficacy has been firmly demonstrated in
                    polycythemia vera (PV) by the ECLAP randomized
                                                                                    us
                                                                                should be paid to antithrombotic prophylaxis adminis-
                                                                                tered to MPN patients with contraindications to ASA,
                                                                                and ad-hoc studies are needed.
                    clinical trial (RCT); however, ASA is also employed in
                                                                                al
                    pts with essential thrombocythemia (ET) and myelofi-
                                                                        ci

                    brosis (MF). Nevertheless, about 2% of the general pop-
                    ulation suffers from ASA intolerance, and about 5% of
                                                                  er


                    ASA users complain of gastric discomfort and gastroin-
                    testinal bleeding. Thus, many MPN pts are treated with
                                                             m



                    anti-P2Y12 agents, mostly ticlopidine (TKL) and clopi-
                    dogrel (CPG). Monotherapy with CPG has been report-
                                                 om




                    ed to be more effective and safe than ASA for antithrom-
                    botic secondary prophylaxis. No comparative data are
                    available for MPN pts. Methods: We examined the
                                              -c




                    medical records of 1278 MPN pts diagnosed at our
                    Center from 1978 to 2021; 1222 were genotyped for
                                      on




                    JAK2V617F, CALR, and MPL mutations. We recorded
                    arterial and venous thrombosis and major bleedings. We
                    computed the observation pt-years under continued           Figure 1.
                                  N




                    monotherapy with ASA 100 mg once daily (od) or
                    TKL/CPG. We excluded pts not receiving antithrombot-
                    ic prophylaxis (n=38), without complete data (n=68),        OC100
                    receiving oral anticoagulants (VKA or DOACs)
                    (n=119), or receiving ASA <100 mg od or other
                    antiplatelet drugs or dual antiplatelet treatment (n=19).   ARTERIAL AND VENOUS THROMBOSIS IN
                    Moreover, we excluded the pts receiving >2 lines of         PATIENTS WITH PHILADELPHIA-NEGATIVE
                    antithrombotic treatment where the time estimate under      MYELOPROLIFERATIVE NEOPLASMS: A SINGLE-
                    ASA or TKL/CPG was fragmented (n=241). In patients          CENTER RETROSPECTIVE ANALYSIS OF 577
                    having received only ASA or TKL/CPG during their            CONSECUTIVE PATIENTS
                    clinical history, we estimated the thrombosis- and bleed-
                    ing-free survival according to the Kaplan Meier method;     E. Morsia1, E. Torre1, F. Marrara1, I. Battilà1,
                    pts were censored at the time of death, thrombosis,         G. Tarantino2, G. Svegliati Baroni2, A. Olivieri1,
                    bleeding, or the last visit. The incidence rate (IR) per    S. Rupoli1
                    100 pt-years was also estimated, dividing the number of
                    thrombotic events for the total observation years.          1Clinica di Ematologia, Ospedali Riuniti di Ancona;
                    Results: We included in the study 793 pts (M/F 339/454,     2Clinica di Gastroenterologia, Ospedali Riuniti di
                    median age 59, range 16-93). The diagnosis was PV in        Ancona, Italy




                                                             [Hematology Reports 2021; 13:s3]                                                [page 69]
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         Oral communications




                     Background and Aims: Arterial thrombotic events and           look for an underlying MPN, particularly in young
                     venous thromboembolism (VTE) are the most common              patients with splenomegaly and high platelet level.
                     causes of morbidity and mortality in the myeloprolifer-
                     ative neoplasms (MPN). MPN are the leading systemic
                     cause of non-provoked VTE in unusual site, including          OC101
                     splanchnic vein thrombosis (SVT) and cerebral vein
                     thrombosis (CVT). This subgroup of patients has been
                                                                                   IDIOPATHIC ERYTHROCYTOSIS WITH
                     showed to have distinct characteristics compared to
                     MPN patients with VTE in usual site, including deep           THROMBOTIC COMPLICATIONS HAS AN ALTERED
                     vein thrombosis and pulmonary embolism. The aim of            BIOMOLECULAR PROFILE
                     this study is analyzed clinical features, molecular signa-
                     ture and outcome data in a real-life single-center cohort     A. Benetti, I. Bertozzi, E. Cosi, G. Biagetti, M.L. Randi
                     of MPN patients who have experienced thrombosis,
                     with a particular focus on unusual site VTE. Materials        First Clinical Medic, Department of Medicine - DIMED,
                     and Methods: In our study, we retrospectively ana-            University of Padova, Italy
                     lyzed a cohort of 577 consecutive patients with MPN
                     who referred to our institute between 2009 and 2020.          Background and Aims: Polycythemia vera (PV), a
                     JMP® Pro 14.0.0 software from SAS Institute, Cary,            myeloproliferative neoplasm, is characterized by a con-
                     NC, USA, was used for all statistical analysis. Results:      sistent risk of hemorrhagic and thrombotic complications,




                                                                                                           ly
                     Among all the MPN population, 19.58% of patients              considered mainly due to rheological alterations induced
                     (n=113) experienced a vascular event during a median          by the excess of red blood cells (RBC). A diagnosis of




                                                                                                   on
                     follow up of 94.3 months (range, 2.4-416.0). According        idiopathic erythrocytosis (IE) is ruled out when all the
                     to WHO 2016 criteria, diagnosis of Polycythemia Vera,         known causes of erythrocytosis are excluded after a com-
                     Essential Thrombocythemia, Primary Myelofibrosis,             plete and accurate diagnostic process. Surprisingly, in a




                                                                                              e
                     and MPN-unclassified was made in 26 patients, 54              recent paper, the estimated prevalence of thrombotic
                     patients, 28 patients, and 5 patients, respectively. In our
                     cohort, only 29 patients presented arterial thrombotic
                     events, including myocardial infarction (45%), stroke
                                                                                       us
                                                                                   complications in IE resulted about half than in PV
                                                                                   patients, in spite of the high hematocrit (HT) levels. The
                                                                                   availability of next generation sequences (NGS) and of a
                                                                                   specific genes panel to search molecular new alterations
                                                                                   al
                     (41%), and peripheral arterial disease (14%). Usual and
                     unusual site venous thrombosis occurred in 41% and            in patients with unexplained erythrocytoses prompted us
                                                                           ci

                     34% in MPN patients with thrombosis, respectively.            in comparing patients’ clinical data with their molecular
                     From the 38 MPN patients with unusual site VTE, 27            genotype. Methods: A complete biomolecular study was
                                                                     er


                     patients had an SVT and 11 a CVT. In this setting of          available in 122 patients (102 males and 20 females,
                     patients, the driven mutation was JAK2V617F in 79%            mean age 53.5±17.2 years). DNA was extracted by gran-
                                                               m



                     with a median allelic burden of 20% using NGS, fol-           ulocytes of patients’ blood samples using EuroGold
                     lowed by CALR in 13% of patients. Compared to those           Blood DNA Mini Kit Plus (EuroClone) according to the
                                                   om




                     with usual site VTE, MPN patients with SVT and CVT            manufacturer’s instructions and stored at -20°C. DNA
                     are younger (ORR 0.87 (0.82-0.93), pValue <0.0001),           concentration was determined by Quibit 4 Fluorometer
                     with higher splenomegaly rate (ORR 0.97 (0.90-0.99),          with QubitTM 1X dsDNA HS Assay Kits (Thermo Fisher
                                                                                   Scientific). The coding sequencing of genes (EGLN1,
                                                -c




                     pValue=0.0003), and with higher PLT count at diagno-
                     sis (ORR 0.06 (0.01-0.38), pValue=0.0085). Unlike             EPAS1, VHL, EPOR) well known to be involved in ery-
                                        on




                     usual site VTE, unusual site VTE seems to presented as        throcytosis were studied with NGS. The panel is an “On
                     the first symptom of disease in about half of cases,          Demand AmpliSeq Panel” designed from the Illumina
                     revealing an underlying MPN (pV<0.001). Long-term             Design Studio platform and validated in silico by
                                   N




                     oral anticoagulation with vitamin K-antagonists (VKA)         Illumina. Results: At the time of the study, in our 122
                     was adopted in 71% of SVT and CVT patients due to             patients, RBC were 5.67±0.3 x 1012/L, hemoglobin (Hb)
                     the MPN-related permanent prothrombotic state, but            172.5±10.5 g/L and HT 50.75±2.56%. No patients had
                     only 32% of MPN patients with usual site VTE was              altered white blood cells (WBC) or platelet (Plts) number.
                     treated with VKA. Survival data in our population             Mean serum erythropoietin (EPO) levels was 10.7±7
                     reported an overall survival (OS) of 238 months (95%          U/L. In 22 (18%) patients EGNL1, 3 (2.4%) VHL, 10
                     I.C. 177-336), poorer in patients with arteriosus events      (8.2%) germinal JAK2 and 7 (5.7%) EPAS1 mutations
                     compared to VTE (usual and unusual sites;                     were found. Only in 4 cases (3.3%) a major thrombotic
                     pValue=0.017). (Figure 1) Older age (HR 1.06 (1.02-           event was reported and the Table 1 contains their main
                     1.84), pValue=0.042) and level of hemoglobin at time          data. Conclusions: The prevalence of thrombosis
                     of thrombosis (HR 0.47 (0.39-0.76), pValue=0.004) sig-        observed in our cohort is lower than the once reported in
                     nificantly influence survival in MPN patients with            the literature (Am. J. Hematol. 2017;92:E639-E641).
                     unusual site VTE. Finally, during the follow-up only          Even if few patients in this cohort had thrombotic com-
                     few patients experienced recurrence thrombosis with a         plications, half of them carry mutations in genes involved
                     recurrence thrombosis free survival rate at 5 years of        in erythrocytosis suggesting that the presence of bio-mol-
                     85%. Conclusions: Our results confirm the thrombotic          ecular alterations may have a correlation with the throm-
                     rate in MPN patients already reported in literature and       botic risk. One patient out of the four with VHL muta-
                     highlight the role of unusual site thrombosis as clue to      tions had a myocardial infarction (MI); VHL mutations




         [page 70]                                          [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 71



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    are known to be associated with cerebral-vascular events      ment (cases). Incident cancer cases were ascertained by
                    and tumor developing while, in contrast, our patient had      direct linkage with the hospital discharge form. The most
                    a MI. The germline mutations in JAK2 gene has been            frequent tumor site was breast (45%) in females and
                    reported to precede the acquisition of PV, JAK2V617F          prostate (25%) in males, followed by non-melanoma skin
                    typical somatic mutation, and thrombotic complications        (11%) and colon-rectum (8%) cancers. From the same
                    may precede PV overt diagnosis: we will strictly follow       HYPERCAN cohort, we extracted a group of 286 ran-
                    this patient in the suspicion of development of PV. Larger    domly cancer-free subjects (154M/132F, 53 years) to
                    studies are needed to confirm this observation.               perform a case-control study. Hemoglobin, hematocrit,
                                                                                  tPA, and FVIII levels at T0 were significantly higher in
                    Table 1.                                                      cases compared to controls. Logistic regression analysis
                                                                                  corrected for gender showed that high tPA levels at T0
                                                                                  were significantly associated with cancer occurrence
                                                                                  (OR=1.052, p=0.039). By ROC curve analysis
                                                                                  (AUC=0.563) a cut-off of 5.62 ng/mL of tPA was estab-
                                                                                  lished to predict cancer occurrence. Logistic regression
                                                                                  analysis corrected for age, smoke habits, alcohol intake,
                                                                                  BMI, and hypertension revealed that a tPA level >cut-off
                    OC102                                                         was an independent risk factor for cancer occurrence
                                                                                  (OR 1.70; 95% CI:1.04-2.77; p=0.034). In the same




                                                                                                           ly
                                                                                  cohort of case-control donors, we also compared the
                    HEMOSTATIC BIOMARKERS MAY PREDICT AN




                                                                                                   on
                                                                                  variation in the hemostatic biomarker levels between T0
                    INCREASED RISK OF CANCER OCCURRENCE IN                        and T1 in relation to cancer risk. We found that only D-
                    HEALTHY SUBJECTS: RESULTS FROM THE                            Dimer levels were significantly (p=0.023) different
                    HYPERCAN STUDY                                                between the two-time points. An increase of one or more




                                                                                             e
                                                                                  quartiles in D-dimer levels from T0 to T1 was signifi-
                    S. Gamba1, M. Marchetti1, L. Russo1, C. Giaccherini1,
                    S. Bolognini1, C.J. Tartari1, C. Verzeroli1, C. Ticozzi1,
                    A. Vignoli1, F. Schieppati1, G. Sampietro2,
                                                                                      us
                                                                                  cantly associated with cancer occurrence in a model of
                                                                                  logistic regression analysis corrected for age, smoke
                                                                                  habits, alcohol intake, BMI, hypertension (OR 1.80; 95%
                                                                                  al
                    P. Malighetti3, D. Spinelli3, A. Falanga1,4                   CI:1.05-3.01; p=0.032). Conclusions: Our study shows
                                                                                  that among the different hemostatic parameters analyzed,
                                                                          ci

                    1Department  of Immunohematology and Transfusion              tPA and D-dimer seem to be potential biomarkers to iden-
                    Medicine, Hospital Papa Giovanni XXIII, Bergamo,              tify subjects at increased cancer risk. In particular, higher
                                                                    er


                    Italy; 2Unit of Epidemiology and Statistics, ATS              levels of tPA, measured at enrollment, are significantly
                    Bergamo, Italy; 3Department of Management,                    associated with cancer occurrence. In addition, the pre-
                                                              m



                    Information and Production Engineering, University of         dictive capacity of higher D-dimer levels is linked to the
                    Bergamo, Bergamo, Italy; 4Department of Medicine and          possibility to measure its variation over time.
                                                  om




                    Surgery, University of Milano-Bicocca, Monza, Italy

                    Background and Aims: Malignancy induces a hyperco-            OC103
                                               -c




                    agulable state in the host, that may represent the initial
                    manifestation of occult cancer. The aim of this study is to
                                                                                  VENOUS THROMBOEMBOLISM (VTE) IN
                                       on




                    establish whether, in a large cohort of healthy blood
                    donors from the Bergamo area enrolled in the HYPER-           PATIENTS WITH HODGKIN LYMPHOMA:
                    CAN study, a hypercoagulable state or its persistence         MONOCENTRIC STUDY OF THROLY AND
                                  N




                    may predict early cancer diagnosis. Methods: The              KHORANA RISK SCORES
                    HYPERCAN study is a prospective Italian, multicenter
                    observational study specifically designed to evaluate         G.M. Assanto, A. Serrao, S. Pontecorvo, F. Malfona,
                    whether hemostatic biomarkers may be an innovative            G.M. D’Elia, G. Annechini, A. Pulsoni, I. Del Giudice,
                    tool for risk assessment, early diagnosis, and prognosis in   A. Chistolini
                    cancer. From 2012 to date, 10,294 healthy blood donors
                    (72% males; median age: 47 years) have been prospec-          Hematology, Department of Translational and Precision
                    tively enrolled in the study and followed up for at least 5   Medicine, Sapienza University of Rome, Rome, Italy
                    years for cancer occurrence. Blood samples were collect-
                    ed at enrollment (T0) and after 6-18 months (T1), togeth-     Background and Aims: Venous Thromboembolism
                    er with clinical, hematological, and lifestyle question-      (VTE) affects the morbidity of Hodgkin Lymphoma
                    naires. Search for malignant tumors was conducted every       (HL) patients. Thrombosis Lymphoma (ThroLy) and
                    6 months. Plasma D-dimer, FVIII, fibrinogen, tissue           Khorana scores, established for lymphomas and cancer
                    plasminogen activator (tPA), and plasminogen activator        patients, can be applied to HL to assess the basal risk of
                    inhibitor-1 (PAI-1) were measured. Results: During a          VTE, due to heterogeneous thrombotic risk; currently
                    median follow-up of 5.9 years, 217 blood donors devel-        there is no indication to perform thrombosis prophylaxis.
                    oped cancer, 190 (122M/68F, 54 years) of them received        The study’s aim was to validate prognostic scores and to
                    a cancer diagnosis after at least 6 months from enroll-       assess clinical factors, which could improve their predic-




                                                               [Hematology Reports 2021; 13:s3]                                                   [page 71]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 72



         Oral communications




                     tive power. Methods: Retrospective monocentric cohort       OC104
                     study, we evaluated patients with newly diagnosed HL
                     (from 2018 to 2020). A total of 134 patients were includ-
                     ed to validate these scores and evaluate other potential    THE IMPACT OF HEMATOCRIT ON
                     risk factors in terms of thrombotic event rate and throm-   POSTOPERATIVE BLEEDING RISK IN PATIENTS
                     botic-free survival. The median age at diagnosis was 42     UNDERGOING ROBOTIC RADICAL
                     years (range 18-85), and median follow-up was 23            PROSTATECTOMY
                     months (range 6-49), thrombotic scores were validated in
                     univariate and multivariate analysis for categorical and    S. Arcudi1, A. Artoni1, E. De Lorenzis2, C. Silvani2,
                     continuous variables as an association to primary end-      F. Spanu3, S. Villa3, M. Capecchi1, E. Montanari2,
                     points. Results: According to Khorana criteria, interme-    F. Peyvandi1
                     diate-risk (IR) were 80.2% (n=105), high-risk (HR) were
                                                                                 1Angelo  Bianchi Bonomi Hemophilia and Thrombosis
                     19.8% (n=26). Regarding ThroLy criteria, low-risk (LR)
                     patients were 16.7% (n=22), IR were 57.6% (n=76) and        Center, IRCCS Ca’ Granda Ospedale Maggiore
                     HR were 25.8% (n=34). All patients received first-line      Policlinico di Milano, Milan, Italy; 2Urology Unit,
                     therapy, and 19 (14.4%) subsequently experienced            Fondazione IRCCS Ca’ Granda Ospedale Maggiore
                     relapse/progression. Central venous catheter (CVC) was      Policlinico, Milan, Italy; 3Clinical Laboratory,
                     implanted in 42.9% (n=57). No patient had previous          Fondazione IRCCS Ca’ Granda Ospedale Maggiore
                     thrombotic events. A total of 22 thrombotic events were     Policlinico, Milan, Italy




                                                                                                         ly
                     observed after a median follow-up of 20 months (range
                     1-45), 10 were CVC-related. The presence of CVC was         Background and Aims: Robot-assisted radical prosta-




                                                                                                 on
                     related to a higher incidence of thrombotic events, 23%     tectomy (RARP) represents the first-choice surgical pro-
                     against 10.8% thrombotic rate in patients without (p        cedure for the treatment of localized prostate cancer. The
                     0.049, OR 1.6 0.90-2.79). Khorana HR group showed an        effects of blood parameters and blood group on the




                                                                                            e
                     increased rate of thrombotic events compared to IR, 28%     bleeding risk had never been investigated. Given this
                     (7/25) vs 11.7% (12/103) respectively (p 0.045). A robust
                     association was found between ThroLy risk categories
                     and thrombotic risk: among HR patients, VTE was
                                                                                     us
                                                                                 background, our study aims to investigate whether blood
                                                                                 parameters and/or blood group may influence the bleed-
                                                                                 ing risk in patients undergoing RARP. Methods: We
                     observed in 32.4% (11/34), against 10.5% (8/76) and
                                                                                 al
                                                                                 conducted a prospective cohort study enrolling consecu-
                     4.55% (1/22), for IR and LR respectively, with an esti-     tive patients undergoing RARP in our hospital, from
                                                                         ci

                     mated thrombotic EFS of 30 months for HR vs 44 for LR       January 2017 to December 2020. Patients were evaluat-
                     (p 0.002 Figure IA). Prophylaxis with enoxaparin abro-      ed with full blood count before and after surgery.
                                                                   er


                     gated the thrombotic risk in HR patients (p 0.56).          Demographic and clinical data were collected with par-
                     Hypertension (HTA) showed a significant association to      ticular attention to other known risk-factors for RARP-
                                                              m



                     thrombotic events, other factors did not show a signifi-    associated bleeding (prostate volume, BMI, smoking
                     cant association. Among hypertensive patients, a higher     status, nerve sparing technique). Bleeding was assessed
                                                  om




                     rate of VTE was observed with 29.6% (8/27) as opposed       both as direct quantification of blood loss during surgery
                     to 12.6% (13/103) in non-hypertensive patients (p           and as the difference in hemoglobin values pre- and
                     0.038). HTA (HR 2.44, p 0.04 CI1.01-6) and HR ThroLy        post-surgery. Linear regression analyses were per-
                     (HR 2.81, p 0.009 CI1.2-6) were confirmed on multivari-
                                               -c




                                                                                 formed, corrected for confounders and known risk fac-
                     ate analysis. We then defined a new risk category which     tors for RARP-associated bleeding, and risk estimates
                     included HR ThroLy patients with HTA, which showed
                                       on




                                                                                 were evaluated with the method of Woolf. Results: In
                     inferior estimated EFS vs other HR patients (21 vs 35       the study period 191 consecutive patients were enrolled;
                     months) with an incidence of 50% of VTE events (6/12)       mean age was 67.8 years. Mean blood loss was 366.2
                                   N




                     vs 22.8% of conventional HR (5/22) (p 0.001 Figure IB).     mL and mean hematocrit pre-surgery was 42.9%. Table
                     Conclusions: ThroLy score proved its utility in VTE-risk    1 shows the pre-surgical clinical characteristics of the
                     assessment, HTA could be integrated to augment ThroLy       191 patients enrolled. Continuous variables are
                     predictive power improving the management of VTE in         expressed as mean, range and standard deviation (SD).
                     HL, thrombotic prophylaxis should be addressed to high-     When analyzing the basal hematocrit with linear regres-
                     risk categories.                                            sion, a strong negative correlation was seen with the dif-
                                                                                 ference in hemoglobin pre- and post-surgery (p<0.001).
                                                                                 The relationship remained strong after correction for
                                                                                 age, smoke, BMI, prostate volume and nerve sparing
                                                                                 (p<0.001). Pre-surgery hemoglobin levels were less
                                                                                 associated with an increased bleeding risk (p=0.07),
                                                                                 while PT, aPTT and platelet count did not influence sur-
                                                                                 gical losses. Individuals carrying the blood group O
                                                                                 were not at increased risk for higher blood loss
                                                                                 (OR=0.76; 95%CI 0.4-1.4; p=0.4). Conclusions: A
                                                                                 lower basal hematocrit is associated with an increased
                                                                                 bleeding during RARP. It can be hypothesized that
                     Figure 1.                                                   patients with higher hematocrit may be more hemostatic




         [page 72]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 73



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    competent since platelets are radially pushed near the      phylaxis in the remaining 94 patients who had Khorana
                    vessel’s wall, as previously described in other settings.   score <2 were: previous VTE (6 patients), other VTE
                                                                                risk factors (31), cancer-related risk factors (34), throm-
                    Table 1.                                                    bophilia (5), multiple indications (18). Patients receiving
                                                                                thromboprophylaxis had a worse performance status (p
                                                                                0.03). At 6 months, VTE occurred in 32/443 (7.2%)
                                                                                patients, of whom 12/206 (5.9%) on thromboprophylax-
                                                                                is and 20/227 (8.9%) not receiving thromboprophylaxis
                                                                                (p 0.27). The same comparison for major bleeding was
                                                                                3.9% and 2.6% (p 0.45) and for death 4.4% and 0.4% (p
                                                                                0.01). Conclusions: In real-world clinical practice set-
                                                                                ting, pharmacological thromboprophylaxis choice was
                                                                                based on Khorana score or other clinical determinants.
                                                                                This strategy was associated with similar VTE incidence
                    OC105                                                       as compared to clinical trials.

                    PHARMACOLOGICAL THROMBOPROPHYLAXIS                          Table 1. Baseline characteristics.
                    TO PREVENT VENOUS THROMBOEMBOLISM IN
                    AMBULATORY PATIENTS WITH CANCER




                                                                                                           ly
                    UNDERGOING ANTINEOPLASTIC DRUGS IN REAL




                                                                                                   on
                    WORLD CLINICAL PRACTICE: A COHORT STUDY

                    M.P. Donadini1, A. Squizzato2, P. Ferrazzi3, C. Secco3,
                    S. Maggioni4, C. Lodigiani5




                                                                                            e
                    1Department   of Medicine and Surgery, University of
                    Insubria and ASST Sette Laghi, Varese, Italy; 2Research
                    Center on Thromboembolic Disorders and
                                                                                    us
                                                                                al
                    Antithrombotic Therapies and ASST Lariana, University
                    of Insubria, Como, Italy; 3IRCCS Humanitas Research
                                                                        ci

                    Hospital, Rozzano, Italy; 4School of Medicine,
                    University of Insubria, Varese, Italy; 5Center for
                                                                  er


                    Thrombosis and Hemorrhagic Diseases, IRCCS
                    Humanitas Research Hospital, Rozzano, Milan, Italy
                                                             m




                    Background and Aims: Venous thromboembolism
                                                 om




                    (VTE) is the second leading cause of death in cancer
                    patients. Clinical prediction models can identify ambu-
                    latory patients undergoing antineoplastic drugs who may
                                              -c




                    benefit from primary pharmacological thromboprophy-
                    laxis. Evidence is scant on determinants of thrombopro-
                                      on




                    phylaxis and outcomes in clinical practice. Aim of the
                    study was to estimate the incidence of VTE in a cohort
                    of cancer ambulatory patients undergoing antineoplastic
                                  N




                    drugs, who were systematically assessed for VTE phar-       OC106
                    macological thromboprophylaxis. Methods: In a retro-
                    spective, observational cohort study, based on a stan-
                    dardized protocol being active at the Humanitas             ACUTE ISCHEMIC STROKE: TRANSCRIPTOMICS
                    Research Hospital from 2012, consecutive adult ambu-        AS AN ALLURING APPROACH TO
                    latory patients starting antineoplastic drugs were evalu-   UNDERSTANDING THE PATHOPHYSIOLOGY AND
                    ated for VTE thromboprophylaxis (low molecular              CLINICAL OUTCOMES
                    weight heparin 80 UI/Kg) based on Khorana score and
                    clinical judgment. The primary outcome was sympto-          G. Cassioli1, A. Kura1, A. Sodero2, E. Sticchi1, A. Magi1,
                    matic VTE and asymptomatic deep vein thrombosis at          S. Suraci1, R. De Cario1, A. Consoli3, A. Rosi3,
                    ultrasound performed at 6 months. The study was             S. Nappini3, L. Renieri3, N. Limbucci3, B. Piccardi2,
                    approved by the local institutional review board.           F. Arba2, C. Sarti2, D. Inzitari2, S. Mangiafico3,
                    Results: 443 patients were enrolled up to November          R. Marcucci1, A.M. Gori1, B. Giusti1
                    2019. Baseline characteristics are described in Table 1.
                    Among 206 patients (46.5%) who were given thrombo-          1University of Florence, Department of Experimental
                    prophylaxis, 112 (54.4%) had Khorana score ≥2, as           and Clinical Medicine, Florence; 2AOU Careggi, Stroke
                    compared to 30/227 (13.2%) who did not receive throm-       Unit, Florence; 3AOU Careggi, Interventional
                    boprophylaxis (p <0.01). Indications for thrombopro-        Neuroradiology, Florence, Italy




                                                             [Hematology Reports 2021; 13:s3]                                                 [page 73]
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         Oral communications




                     Background and Aims: Acute ischemic stroke (AIS) is          also suggested novel pathways to be explored that may
                     a complex multifactorial disease characterized by a          provide an important starting point for expanding
                     high rate of morbidity and mortality and represents one      knowledge on ischemic stroke disease.
                     of the leading causes of death and disability worldwide.
                     Current treatments are focused on recanalization of the
                     occluded cerebral vessel via intravenous thrombolysis        OC107
                     with rt-PA and/or endovascular mechanical thrombecto-
                     my (MT). The latter approach created a golden ticket
                                                                                  CIRCULATING MIRNAS RELEASE PREDICTS
                     from a research perspective, supplying new study mate-
                     rial that enabled histopathological analysis of cerebral     SUBOPTIMAL RESPONSE TO ASPIRIN IN
                     thrombi (CT), which not only provided important evi-         PATIENTS AT HIGH CARDIOVASCULAR RISK
                     dence on their cellular composition, but also represents     WITH AND WITHOUT TYPE 2 DIABETES MELLITUS
                     a key pillar for the creation of virtual predictive models
                     of different gene expression. The aim of the study is        R. Liani1,2, P.G. Simeone1,2, S. Ciotti, R. Tripaldi1,2,
                     therefore to investigate global gene expression profiles     A. Boccatonda2,3, D. D’Ardes2,3, A. Recchiuti4,
                     of both CT and peripheral venous blood (PB), not only        M. Romano4, F. Cipollone1,2,3, F. Santilli1,2,3
                     highlighting and exploring whether PB profiles mir-
                                                                                  1Center for Advanced Studies and Technology (CAST),
                     rored CT profiles, but also focusing on the identifica-
                     tion of potential markers of different pathophysiologi-      Chieti University, Chieti, Italy; 2Haemostasis and




                                                                                                           ly
                     cal mechanisms of AIS and/or determinants of clinical        Thrombosis Unit CAST, Chieti University, Chieti, Italy;
                                                                                  3Internal Medicine, Chieti University, Chieti, Italy;
                     outcomes, such as haemorrhagic transformation, 24h




                                                                                                   on
                                                                                  4Department of Medical and Oral Sciences, Italy
                     edema, modified 3 months Rankin scale-mRS, death.
                     Methods: We performed gene expression profiles of
                     RNA samples obtained from 40 CT and 37 PB of 52              Background: The recovery rate of platelet COX-1 activ-




                                                                                             e
                     patients. The CT obtained during MT were stored in           ity during the 12 to 24h dosing interval of aspirin admin-
                     RNA later, while PB, collected before and 24 hours
                     after MT, in tubes containing a reagent that protects
                     RNA from degradation and minimizes ex vivo changes
                                                                                      us
                                                                                  istration, in aspirin-treated subjects, is characterized by
                                                                                  substantial interindividual variability. Circulating
                                                                                  myeloid-related protein (MRP)-8/14 is an inflammatory
                                                                                  protein associated with residual thromboxane (TX)-
                                                                                  al
                     in gene expression. RNA was then extracted by PAX
                     gene blood miRNA kit, the global gene expression pro-        dependent platelet activation in aspirin-treated patients
                                                                          ci

                     file was assessed by Affymetrix technology using             with acute coronary syndrome. Aims: To identify any
                     GeneChip Human Transcriptome Array 2.0, allowing             circulating miRNAs associated with a suboptimal ASA
                                                                    er


                     the analysis of 44,699 genes, with more than 285,000         response in patients at high cardiovascular (CV) risk.
                     full-length transcripts coverage. Data analysis was then     Methods: Two-hundred high CV risk patients (100 with
                                                              m



                     performed in R environment with dedicated pipelines.         type 2 diabetes mellitus (T2DM)) in chronic treatment
                     Results: Data processing and the application of appro-       with ASA (100 mg/day), for cardiovascular prevention,
                                                  om




                     priate filtering criteria showed an average of analyzable    were enrolled. Blood sampling was performed at 10
                     probe sets of 440,085 in CT and 602,874 in PB. In the        (T10) and 24 hours (T24) after a witnessed ASA admin-
                     two different type of specimens 20,341 were found to         istration. Patients were stratified in tertiles according to
                                               -c




                     be common features, whereas 3 and 562 symbols were           serum TXB2 slope. First vs. third tertile were compared.
                     unique in CT and PB, respectively. The Gene Ontology         Circulating miRNAs custom array cards were applied to
                                        on




                     (GO) enrichment analysis allowed the identification of       assay the expression levels of 14 miRNAs in plasma. We
                     the biological processes, common and peculiar, in CT         also measured plasma myeloid-related protein (MRP)-
                     and PB, indicating that peripheral and local mecha-          8/14 as an inflammatory index and predictor of cardio-
                                   N




                     nisms of damage and response to damage are present in        vascular events. Results: miRNA-21-5p (p=.017), 22-5p
                     both. The significance analysis of microarrays, accord-      (p=.026), 24-3p (p=.020), 150-5p (p=.026), 155-5p
                     ing to different outcomes and GO analysis, brought into      (p=.007), 181b-5p (p=.011), 223-5p (p=.021) were sig-
                     focus 221 significant biological processes associated        nificantly lower in first vs. third tertiles at 24 hours after
                     with poor outcome according to mRS in CT, and 27             ASA administration in all patients (Figure 1). MRP-8/14
                     terms associated with 24h edema in PB. Among signif-         were higher in third vs. first sTXB2 slope tertile in all
                     icant terms in CT, those associated with regulation of       patient. MRP-8/14 was directly correlated with miRNA-
                     neutrophil mediated immunity and activation play a           21-5p (rho=.279, p=.008), 22-5p (rho=.264, p=.012), 24-
                     crucial role. Concerning PB, particularly significant        3p (rho=.239, p=.023), 150-5p (rho=.236, p=.025), 155-
                     enriched terms were associated with regulation and           5p (rho=.270, p=.011), 181b-5p (rho=.240, p=.023) and
                     activation of transcriptomes of cells. Conclusions: Our      223-5p (rho=.244, p=.030) in all patients (data not
                     results provided interesting insights into the mecha-        shown). Conclusions: MRP 8/14 may contribute to cir-
                     nisms underlying the AIS and the response to treat-          culating miRNA release and response variability to ASA.
                     ments. In particular, the analysis of CT and PB gene         Vice versa, shorter duration of aspirin effect at 24 hours
                     expression profiles, differentially expressed probe sets     in third sTXB2 slope tertile patients translates into higher
                     and their biological processes alterations according to      degree of TX-dependent platelet activation, possibly pro-
                     stroke outcomes, has not only confirmed and extended         moting the release of both circulating MRP8/14 and
                     several known pathophysiological mechanisms, but             miRNAs.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 75



                                                                       XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                               types, and to VWF antigen levels. Results: α-HL -
                                                                               Both the LDLR c.1773 TT (Beta-coefficient 0.329,
                                                                               p=0.047) and ASGR2 c.-95 TT (Beta-coefficient -
                                                                               0.432, p=0.013) were found to contribute to α-HL,
                                                                               with the ASGR2 association predicted independently
                                                                               from ABO genotypes. These data point out genetic
                                                                               components increasing (ASGR2 TT, 3.60h±0.94SEM)
                                                                               or decreasing (LDLR TT, 0.42h±0.16SEM) α-HL
                                                                               through FVIII/VWF-receptors interactions occurring
                                                                               in the initial FVIII distribution phase. These informa-
                                                                               tion may be relevant for FVIII concentrate dosing in
                                                                               on-demand treatment of bleeding. CLEARANCE - No
                                                                               genetic component remained a significant and inde-
                                                                               pendent contributor of Clearance, which may point out
                                                                               multiple interactions partially competing each other.
                    Figure 1.                                                  Since infrequent homozygous conditions appear to
                                                                               contribute a large proportion of variance and unfavor-
                                                                               able clearance (CLEC4M rs868875 GG, 4.30 ml/h/kg
                                                                               ±1.70SEM), further investigation is needed in larger




                                                                                                      ly
                    OC108
                                                                               studies. β-HL - The ASGR2 c.-95 T>C genotypes were
                                                                               the most significant contributors (TT vs TC, Beta-




                                                                                               on
                    GENETIC COMPONENTS IN FACTOR VIII                          coefficient -0.548, p=0.009), independently from
                    RECEPTORS FOR INDIVIDUAL AND GENOTYPE-                     LDLR, CLEC4M and ABO genotypes. This model
                    MODELED HEMOPHILIA A TREATMENT                             includes a significant prediction by ABO genotypes




                                                                                         e
                                                                               (Beta-coefficient 0.428, p=0.049). MRT - The ASGR2
                    B. Lunghi1, M. Morfini2, N. Martinelli3, A. Branchini1,
                    S. Linari4, G. Castaman4, F. Bernardi1                        us
                                                                               c.-95T>C genotypes were significant contributors
                                                                               (Beta-coefficient -0.493 p=0.023) independently from
                                                                               LDLR, CLEC4M and ABO genotypes. Since also
                                                                               al
                    1Department     of Life Sciences and Biotechnology,        ABO genotypes were MRT modifier as a trend (Beta-
                    University of Ferrara; 2Italian Association Hemophilia     coefficient 0.407, p=0.075), this analysis points out a
                                                                       ci

                    Centers (AICE), Naples; 3Department of Medicine,           specific glycan-dependent association. Further, the
                    University of Verona; 4Center for Bleeding Disorders,      noticeable differences between ASGR2 genotypes val-
                                                                 er


                    Department of Oncology, Careggi University Hospital,       ues (c.-95TT, 40% longer than in c.-95TC) deserve to
                    Florence, Italy                                            be explored for individual and genotype-based modu-
                                                            m



                                                                               lation of prophylaxis regimens. Conclusions: With the
                    Background and Aims: Age, von Willebrand factor            limitation of the small number of HA patients, these
                                                om




                    (VWF) levels and the ABO blood group all together          novel observations may reflect different FVIII-scav-
                    explain only 30% of the large inter-patient variability    enging properties of receptors “in vivo” and provide
                    of factor VIII (FVIII) pharmacokinetics (PK) in hemo-      valuable information for individual and genotype-
                                             -c




                    philia A (HA) patients. Gene variation in the numerous     based modulation of on demand/prophylaxis regi-
                    scavenging receptors for circulating FVIII/VWF is          mens. This analysis will also favor comparison of
                                      on




                    candidate to explain an additional portion of variabili-   receptor function in prophylaxis with extended half-
                    ty. We have recently reported genetic components of        life FVIII concentrates.
                    PK variability with standard half-life (SHL) FVIII,
                                  N




                    most of them independent from VWF antigen levels,
                    in i) the low-density lipoprotein receptor (LDLR) and      OC109
                    in the C-type lectin domain family, particularly in ii)
                    the asialoglycoprotein receptor minor subunit gene
                    (ASGR2) and iii) the family 4 member M (CLEC4M).           A MULTI-TRAIT ASSOCIATION ANALYSIS OF
                    The LDLR c.81 CC homozygotes and the ABO                   PLATELET TRAITS AND BRAIN DISORDERS
                    O/CLEC4M rs868875 G-carriers showed fast FVIII             IDENTIFIES NOVEL SUSCEPTIBILITY LOCI FOR
                    clearance, and the ASGR2 c.-95 TC heterozygotes            MAJOR DEPRESSION, ALZHEIMER AND
                    short mean residence time (MRT). Based on differen-        PARKINSON DISEASE
                    tial cell distribution (macrophages and liver sinusoid
                    endothelial cells) and protein/glycan recognition of       A. Tirozzi1, M. Shasa Quiccione1, C. Cerletti1,
                    receptors, we compared the influence of these genetic      M.B. Donati1, G. de Gaetano1, L. Iacoviello1,2,
                    components. Methods: Linear regression models of           A. Gialluisi1,2
                    SHL FVIII two-compartment model PK parameters in
                                                                               1Department of Epidemiology and Prevention, IRCCS
                    31 HA patients (FVIII coagulant activity ≤2 IU/dL).
                    Main parameters (Alpha and Beta HL, Clearance,             NEUROMED, Pozzilli, Italy 2EPIMED Research Center,
                    MRT) were evaluated in relation to the LDLR,               Department of Medicine and Surgery, University of
                    ASGR2, CLEC4M, F8, and ABO blood-group geno-               Insubria, Varese, Italy




                                                             [Hematology Reports 2021; 13:s3]                                            [page 75]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 76



         Oral communications




                     Background: Among candidate neurodegenerative/neu-           OC110
                     ropsychiatric risk-predictive biomarkers, platelet count
                     (Plt), mean volume (MPV) and distribution width
                     (PDW) have been associated with the risk of Major            GENETIC DELETION OF
                     Depressive Disorder (MDD), Alzheimer’s (AD) and              PRENYLCYSTEINEOXIDASE 1 (PCYOX1)
                     Parkinson’s disease (PD), both through epidemiological       IMPAIRS ARTERIAL THROMBOSIS IN MICE
                     and through genomic studies, suggesting partial co-heri-
                     tability. Methods: We exploited these relationships for a    P. Amadio1, C. Banfi2, M. Zarà1, M. Brioschi2,
                     multi-trait association analysis (MTAG), using GWAS          S. Ghilardi2, L. Sandrini1, S.S. Barbieri1
                     associations of three different platelet traits – Plt, MPV   1Unit of Brain-Heart Axis, Centro Cardiologico Monzino
                     and PDW – and three neurodegenerative/neuropsychi-
                     atric disorders, AD, PD and MDD. Gene-based enrich-          IRCCS, 20138Milano, Italy; 2Unit of Functional
                     ment tests were carried out through MAGMA v1.08,             Proteomics, Metabolomics and Network Analysis,
                     while a network analysis of significantly enriched genes     Centro Cardiologico Monzino IRCCS, 20138Milano,
                     was implemented in dbSTRING v11.5. Results: We ana-          Italy
                     lyzed 4,540,326 Single Nucleotide Polymorphisms
                     (SNPs) shared among the analyzed GWAS, observing             Prenylcysteine Oxidase 1 (PCYOX1), an enzyme
                     149 genome-wide significant multi-trait Linkage              involved in the degradation of prenylated proteins, is
                     Disequilibrium independent associations (p<5×10-8) for       expressed in different tissues, including vascular and




                                                                                                         ly
                     AD, 70 for PD and 139 for MDD. Among these, 27 novel         blood cells. The secretome from Pcyox1-silenced cells
                     associations were detected for AD, 34 for PD and 40 for      reduced platelet adhesion to both fibrinogen and




                                                                                                  on
                     MDD. Out of 18,781 genes with annotated variants with-       endothelial cells, suggesting its potential contribution
                     in ±10 kb, 62 genes were enriched for associations with      to thrombosis. In this study, we analysed the contribu-
                     AD, 70 with PD, and 125 with MDD (p<2.7×10-6). Of            tion of PCYOX1 in arterial thrombosis by using a




                                                                                            e
                     these, 7 genes were novel for AD (EPPK1, TTLL1, PAC-         murine model. All procedures were performed on
                                                                                  Pcyox1 knock-out (Pcyox1KO) mice and compared to
                     SIN2, TPM4, PIF1, ZNF689, AZGP1P1), 2 for PD
                     (SLC26A1, EFNA3) and 1 for MDD (HSPH1). The
                     resulting network showed a significant excess of interac-
                                                                                     us
                                                                                  littermate wild type (WT) mice. Ferric Chloride
                                                                                  (FeCl3) arterial injury and pulmonary thromboem-
                                                                                  bolism models have been used to induce thrombosis.
                                                                                  al
                     tions (enrichment p=1.0×10-16) (Figure 1).
                                                                                  The phenotype and the function of platelets were stud-
                                                                                  ied by flow cytometry and functional tests. The pres-
                                                                          ci

                                                                                  ence of PCYOX1 in platelets was assessed by mass
                                                                    er


                                                                                  spectrometry analysis. Thrombus formation after
                                                                                  FeCl3 injury of the carotid artery was delayed in
                                                                                  Pcyox1KO mice, which were also protected from col-
                                                              m




                                                                                  lagen-epinephrine       induced     thromboembolism.
                                                                                  Pcyox1KO mice displayed physiologic blood cell
                                                  om




                                                                                  count, vascular pro-coagulant activity, and plasma fib-
                                                                                  rinogen levels but reduced platelet/leukocyte aggre-
                                                                                  gates in whole blood, as well as platelet aggregation,
                                               -c




                                                                                  alpha granules release and the αIIbβ3 integrin activa-
                                                                                  tion in platelet-rich plasma in response to ADP or
                                        on




                                                                                  TRAP. PCYOX1 was greatly expressed in washed
                                                                                  platelets isolated from WT mice but phosphorylation
                                   N




                                                                                  pathway activation, adhesion ability and aggregation
                                                                                  were similar to those observed in platelets from
                                                                                  Pcyox1KO. The presence of Pcyox1KO mice-derived
                                                                                  plasma, impaired agonists-induced aggregation of WT
                                                                                  mice-derived platelets. Our findings, showing
                                                                                  platelets hyporeactivity and impaired arterial throm-
                                                                                  bosis in the absence of PCYOX1, suggest it as a
                                                                                  potential novel target for antithrombotic therapy.

                     Figure 1.
                                                                                  OC111
                     Discussion: We identified novel genes involved in the
                     organization of cytoskeletal architecture (EPPK1,            FAMILIAL HYPERCHOLESTEROLEMIA: INSIGHT
                     TTLL1, PACSIN2, TPM4), in telomere shortening                INTO GENETIC PREDISPOSITION PROFILE
                     (PIF1), regulation of cellular aging (ZNF689,
                     AZGP1P1) and in neurodevelopment (EFNA3), gaining            E. Sticchi1, A. Kura2, R. De Cario2, G. Barbieri2,
                     novel insights into the underlying biology of neurode-       A. Magi3, S. Suraci2, A.M. Gori1, F. Cesari4,
                     generative/neuropsychiatric disorders.                       G.M. Scaturro5, E. Lotti4, R. Marcucci1, B. Giusti1




         [page 76]                                         [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 77



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    1University of Florence, Department of Experimental           concerns DLCN score, performed in the adult popula-
                    and Clinical Medicine, Atherothrombotic Diseases              tion, significantly higher values in subjects carrying
                    Center, Careggi Hospital, Florence; 2University of            LDLR mutation were found. Conclusions: Present data
                    Florence, Department of Experimental and Clinical             support the involvement of other multiple loci, beyond
                    Medicine, Florence; 3University of Florence,                  LDLR gene, in the modulation of lipid profile, as well as
                    Department of Information Engineering, Florence;              cardiovascular risk. Nevertheless, further expansion of
                    4Atherothrombotic Diseases Center, Careggi Hospital,
                                                                                  genetic analysis to a larger cohort might allow a better
                    Florence; 5Metabolic Disease Unit, AOU Meyer, Italy           comprehension of the role of further major/modifier
                                                                                  genes, as well as of accumulation of common small-
                    Background and Aims: Familial hypercholesterolemia            effect LDL-C raising alleles in determining LDL-C lev-
                    (FH) represents a heritable disorder associated with path-    els and cardiovascular events.
                    ogenic variants in LDLR or APOB or PCSK9 (dominant
                    form), and in LDLRAP1 (recessive form) with increased
                    risk of premature coronary artery disease. Moreover, pre-     OC112
                    vious data showed that in about 60% of patients who are
                    mutation-negative the clinical phenotype can be associat-
                    ed with an accumulation of common small-effect LDL-           DATA MINING FOR PROFILING THE MUTATIONAL
                    C-raising alleles using a 12- Single nucleotide polymor-      LANDSCAPE OF VON WILLEBRAND DISEASE
                    phisms (SNPs) score (Talmud et al. 2013). In the present




                                                                                                         ly
                    study we genetically characterize a cohort of adult and       O. Seidizadeh1,2, A. Cairo1, L. Baronciani1,
                    paediatric FH patients in order to provide further insight    M.T. Pagliari1, S.M. Siboni1, E. Biguzzi1, F. Peyvandi1,2




                                                                                                  on
                    into the genetic predisposition profile. Methods: We          1Fondazione
                    analysed 90 FH patients [adults with clinically                             IRCCS Ca’Granda Ospedale Maggiore
                    possible/probable or definite FH using the most common        Policlinico, Angelo Bianchi Bonomi Hemophilia and




                                                                                            e
                    diagnostic algorithm, Dutch Lipid Clinic Network Score        Thrombosis Center and Fondazione Luigi Villa, Milan,
                                                                                  Italy; 2Department of Pathophysiology and
                    (DLCN)]. Genetic analysis was perfomed through a tar-
                    geted high-throughput (HTS) panel of 57 genes, includ-
                    ing loci involved in lipid metabolism, genes supposed to
                                                                                     us
                                                                                  Transplantation, Università degli Studi di Milano,
                                                                                  Milan, Italy
                                                                                  al
                    be involved in dyslipidemia, pharmacogenetics of statins,
                    genes related to higher susceptibility for the polygenic      Background: Von Willebrand disease (VWD) with a
                                                                          ci

                    forms of FH, HDL and triglycerides related diseases, and      population prevalence of 1% is caused by mutations in
                    regions interested by polymorphisms of the Talmud             the von Willebrand factor (VWF). However, its exact
                                                                    er


                    genetic score. DNA libraries were prepared using Agilent      prevalence is not yet established. More than 900 dis-
                    HaloplexHS enrichment system, and sequencing was              tinct mutations have been found in the VWF so far.
                                                              m



                    assessed by Illumina MiSeq platform. Results: Among           Aims: To explore the VWF mutational burden and to
                    90 patients analyzed, 41 carried a rare variant in LDLR       estimate VWD prevalence in the general population
                                                  om




                    gene, whereas 49 patients were LDLR-negative. Talmud          using the genome Aggregation Database (gnomAD).
                    score evaluation in both groups showed a higher mean          Methods: VWF variants were obtained from the
                    value in patients without LDLR mutations, with respect        gnomAD (v2.1.1; 141,456 subjects) and were com-
                                                                                  pared to the Human-Gene-Mutation-Database (HGMD)
                                               -c




                    to the LDLR-positive group (0.972±0.207 vs
                    0.941±0.175). HTS analysis revealed that 14 LDLR              and Leiden-Open-Variation-Database (LOVD v.2.0 and
                                                                                  v.3.0). The following variants were considered patho-
                                       on




                    mutation-positive patients also carried likely pathogenet-
                    ic/uncertain significance mutations in APOB or LDL-           genic: nonsense and frame-shift; splicing (+2/-2); splic-
                    RAP1 genes. In patients without LDLR mutations, at            ing (+8/-8) and missense predicted deleterious by 3/3
                                  N




                    least 2 rare variants were identified in 24 patients (49%),   and 7/7 tools respectively or being associated with
                    and at least 3 rare variants were identified in 18 patients   VWD in HGMD and LOVD2/3. All identified patho-
                    (37%). In these patients, a total of 117 rare variants with   genic variants were used to evaluate the VWF muta-
                    uncertain significance/conflicting interpretation of path-    tional burden and VWD prevalence. Results: Using
                    ogenicity have been identified in 44 different genes          gnomAD, we identified 505 distinct VWF pathogenic
                    (APOB, PCSK9, LDLRAP1, ABCB1, ABCG2, ABCG5,                   variants (244 unique): 287 not reported (novel) and 218
                    ABCG8, ANGPTL3, APOA4, CELSR2, CETP,                          already reported. 138 of 218 gnomAD variants were not
                    CREB3L3, DAB2, GCKR, GHR, HFE, ITIH4, LCAT,                   predicted as pathogenic by all in-silico tools but were
                    LIPC, LIPI, LMF1, LPA, LPL, LRP1, MTTP, NPC1,                 reported to be associated with VWD in HGMD and/or
                    NYNRIN, PON1, PPP1R17, SCARB1, SLCO1B1,                       LOVD databases. Thirteen variants had a minor-allele-
                    SLC12A4, SREBF1, SREBF2, SLC22A1, EPHX2,                      frequency (MAF) >0.01 in at least one population. In
                    GPD1, OSBPL5, STAP1, ABCA1, DGAT1, INSIG2,                    particular, 3 variants (p.Arg2185Gln; p.Met740Ile;
                    NPC1L1, APOA5). Among patients analyzed, 29 were              p.His817Gln) were very frequent in Africans/Africans
                    younger than 18 yrs. In the adult population, LDL cho-        American (MAF: 0.11-0.18). Due to these variants, the
                    lesterol levels were comparable between LDLR positive         African population was excluded from the calculation
                    and LDLR negative group, whereas in subjects younger          of VWD prevalence in the general population and was
                    than 18 yrs significantly higher LDL cholesterol levels in    analyzed standalone. This analysis revealed a heterozy-
                    the LDLR positive group were observed. Moreover, as           gote frequency of 0.13 among all populations with a




                                                               [Hematology Reports 2021; 13:s3]                                               [page 77]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 78



         Oral communications




                     prevalence of 0.45 and 13.4 for recessively-inherited        istry was created to document and evaluate PLG defi-
                     and autosomal-dominant, respectively among 100 indi-         ciency unmet needs with the goal of define its natural
                     viduals. Conclusions: Through our systematic analysis        history. Methods: Up to 100 probands with their 1st
                     of>140,000 individual exome/genome data from the             degree non-affected family members will be enrolled in
                     gnomAD database, we found 287 novel VWF variants             the 4 -year study. Clinical, genetic, and laboratory data
                     predicted as pathogenic. Our results demonstrated that       are collected at baseline and at semestral visits;
                     the “true” worldwide prevalence of VWD is more than          unscheduled visits are also included (new lesions or
                     10 times higher than that reported so far.                   pregnancy). Specimens are also collected to perform
                                                                                  measurements of PLG activity (PLG:C) and antigen
                                                                                  (PLG:Ag) and to confirm diagnosis by molecular char-
                     OC113                                                        acterization. Therapeutic interventions are recorded
                                                                                  along with efficacy and safety observations. Results:
                                                                                  To date, 11 centers (6 countries) are actively enrolling
                     HYPOPLASMINOGENEMIA: AN INTERNATIONAL                        with 115 subjects entered. Analysis was performed on
                     RETROSPECTIVE AND PROSPECTIVE COHORT                         27 subjects (21 probands, 6 siblings) with complete
                     STUDY (HISTORY): THE INTERNATIONAL                           data available; Table 1 includes demographic data and
                     REGISTRY OF PATIENTS WITH PLASMINOGEN                        most frequently affected areas. Mean PLG:C and
                     DEFICIENCY                                                   PLG:Ag were 15.3% (nr 80 – 132) and 15.8 ug/ml (nor-
                                                                                  mal range 70 – 215), respectively. Eyes and ear lesions




                                                                                                         ly
                     M. Menegatti1, J. Bowen2, M. Boscarino1, C. Nakar2,          were reported early in life, 1 and 3.5 years (median),
                     R. Palla3, S. Siboni1, M. Torres4, M. Batur5,                respectively. Conclusions: HISTORY is the first com-




                                                                                                  on
                     R. Natesirinilkul6, C. Roberson2, H. McDaniel7,              prehensive retrospective/prospective international reg-
                     S. Al8, N. Rodriguez9, T. Celkan10, J. Ceresetto11,          istry to document and evaluate PLGD current knowl-
                     N. Thukral2, B. Hardesty2, F. Peyvandi1,3, A. Shapiro2       edge gaps. This first analysis demonstrates a significant




                                                                                             e
                                                                                  decrease in PLG:C and PLG:Ag levels in affected
                     1Fondazione   IRCCS Ca’ Granda Ospedale Maggiore
                     Policlinico, Angelo Bianchi Bonomi Hemophilia and
                     Thrombosis Center and Fondazione Luigi Villa, Milan,
                                                                                      us
                                                                                  patients, and that severe symptoms including eye and
                                                                                  ear lesions occur early in life. Interestingly, the study
                                                                                  has not yet recorded a subject of African descent.
                     Italy; 2Indiana Hemophilia & Thrombosis Center,
                                                                                  al
                     Indianapolis, United States; 3Università degli Studi di      Table 1.
                                                                          ci

                     Milano, Department of Pathophysiology and
                     Transplantation, Milan, Italy; 4Cook Children’s,
                                                                    er


                     Hematology/Oncology, Fort Worth, United States; 5Van
                     Yuzuncu Yil University, Ophthalmology Department, Van,
                                                              m



                     Turkey; 6Chiang Mai University, Department of
                     Pediatrics, Chiang Mai, Thailand; 7Vanderbilt Children’s
                                                  om




                     Hematology-Oncology, Nashville, United States; 8Dokuz
                     Eylül University Research Clinical Immunology, Izmir,
                     Turkey; 9The University of Texas Health Science Center
                     at Houston, Houston, United States; 10Istanbul University
                                               -c




                     Cerrahpasa, Cerrahpasa Medical Faculty Pediatric
                     Hematology, Istanbul, Turkey; 11Hospital Britanico
                                        on




                     Buenos Aires, Buenos Aires, Argentina
                                   N




                     Background and Aims: Plasminogen (PLG) deficien-
                     cy (PLGD) is a rare inherited chronic disorder affecting
                     ~1.6 per million population. Uncontrolled growth of
                     fibrin-rich pseudomembranes on mucous membranes
                     carries potential mortality risk and may affect eyes, oral
                                                                                  OC114
                     cavity and middle ears, central nervous system and skin
                     as well as respiratory, female genitourinary, gastroin-
                     testinal and renal tracts and is associated with morbidity   IMPACT ON PLATELET FUNCTION OF A NOVEL
                     and mortality. Ligneous conjunctivitis is the most com-      VARIANT OF THE BETA ISOFORM OF THE TBXA2R
                     mon manifestation (~80%) and may lead to sight loss.         GENE ASSOCIATED WITH A HETEROGENEOUS
                     Data regarding management of patients with PLGD is           BLEEDING PHENOTYPE
                     scarce and based on case reports/series and small clini-
                     cal trials. In addition, even though in 2021, the FDA        L. Bury, E. Falcinelli, A.M. Mezzasoma, M. Borghi,
                     approved the first PLG concentrate for such patients,        E. Cesari, P. Gresele
                     specific replacement therapies (PLG ophthalmologic
                     and systemic concentrates) are not currently commer-         Department of Medicine and Surgery, Section of Internal
                     cially available. With this as background, the first com-    and Cardiovascular Medicine, University of Perugia,
                     prehensive retrospective/prospective international reg-      Perugia, Italy




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                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                                dual range. Genetic variants affecting genes involved in
                    Background and Aims: The thromboxane A2 receptor            platelet function were assessed by next generation
                    (TP) plays an essential role in hemostasis by binding       sequencing, performing targeted sequencing of 89
                    thromboxane A2 (TXA2), a potent platelet agonist and a      genes. Results: We show that ultrapurified platelets
                    vasoconstrictor. There are two isoforms of the receptor,    express the TPβ isoform mRNA and protein. Platelet
                    TPα and TPβ, generated by alternative splicing from the     LTA was defective in response to arachidonic acid,
                    TBXA2R gene, which show similar ligand-binding              U46619, U44069 and secondary wave to ADP and epi-
                    activity. While the expression and function of TPα in       nephrine in the patient and in her father. Real time PCR
                    platelets is well established, that of TPβ is debated.      showed a 50% reduction of the mRNA coding for TPβ
                    TXA2 binds to the TP receptor leading to the activation     accompanied by a reduction of the TPβ protein by WB
                    of Gq/11 and G12/13, but also of Gh, Gαs and Gαi.           in patient platelets. ADP- and U46619-induced Ca2+
                    Different coupling to G-proteins of the two isoforms has    mobilization was impaired, suggesting reduced activa-
                    been suggested but actually very few studies have ana-      tion of Gq/11 and Gh. Stimulation with U46619
                    lyzed this aspect. TP-receptor deficiency is an autosomal   decreased cAMP in platelets pre-treated with Iloprost to
                    dominant platelet disorder characterized by mucocuta-       a lower extent than in control platelets (20±2% vs
                    neous hemorrhages caused by variants in TBXA2R. Only
                                                                                47±15% of controls), suggesting impaired activation of
                    6 variants in few families all involving TPα have been
                                                                                Gαi. We did not identify by targeted sequencing rare
                    described. Moreover, the severity of bleeding differs
                                                                                variants in other platelet genes, but found that the patient
                    among patients with the same gene variant for unclear




                                                                                                         ly
                    reasons. We recently found a novel TBXA2R variant           and not the father carries the common rs5758 in
                    (g.A16755T) affecting the alternative splice site of TPβ    TBXA2R, previously associated with reduced platelet




                                                                                                 on
                    in a woman with a lifelong bleeding diathesis and in her    activation in response to U46619. Conclusions: Our
                    father who did not report bleeding symptoms. Aim of         study characterizes the first reported inherited platelet
                    our study was to shed light on the impact of this novel     function defect due to a TBXA2R variant regulating the




                                                                                           e
                    TBXA2R variant on platelet function and to elucidate the    expression of the TPβ isoform associated with a muco-
                                                                                cutaneous bleeding diathesis. We also show that the TPβ
                    function of TPβ in platelets. Methods: TP isoforms
                    expression was assessed by real time PCR and Western
                    blotting (WB) using antibodies selective for the two TP
                                                                                    us
                                                                                defect was associated with impairment of Gq/11, Gh and
                                                                                Gαi activation, suggesting that TPβ is involved in the
                    isoforms. Platelet function was studied by light trans-     regulation of all these pathways similar to TPα. These
                                                                                al
                    mission aggregometry (LTA), flow cytometry and lumi-        findings definitely suggest that TPβ has a role in platelet
                    aggregometry. Ca2+ mobilization induced by various          function. However, defective TPβ requires for a full phe-
                                                                        ci

                    agonists was assessed by flow cytometry using the           notypic expression that other genetic variant(s), like the
                                                                  er


                    FLUO 3-AM dye. cAMP production was assessed using           common rs5758 polymorphism of TBXA2R, are simulta-
                    the cAMP Enzymeimmunoassay Biotrak (EIA) System             neously present.
                                                             m
                                                 om
                                              -c
                                      on
                                  N




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         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                                         ly
                                                                                         on
                                                                                 e
                                                                            us
                                                                      al
                                                                  ci
                                                             er
                                                          m
                                              om
                                           -c
                                     on
                                 N




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                                                                           XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                            Poster




                                                                                                        ly
                                                                                                 on
                    PO001                                                        thrombotic events during hospitalization. At the admis-
                                                                                 sion, 79 patients had CAC criteria and those patients




                                                                                           e
                                                                                 required more often an increase of intensity of medical
                    LONG TERM COVID-ASSOCIATED
                                                                                 care (33.6% vs 22.3%; p 0.019) and had more thrombot-
                    COAGULOPATHY

                    A. Milia1, S. Neroni2, B. Giusti2, A.M. Gori2,
                                                                                     us
                                                                                 ic events (10% vs 5.8% p=0.14). Laboratory parameters
                                                                                 associated with future thrombotic events and need of
                                                                                 increase intensity of medical care during the hospital-
                                                                                 al
                    A. Rogolino2, F. Cesari2, E. Sticchi2, A. Bertelli2,         ization were higher platelets, D-dimer, fibrinogen, C-
                    D. Poli2, R. Marcucci2
                                                                                 reactive protein, ferritin and LDH. At follow up evalua-
                                                                           ci

                    1Internal                                                    tion (median 5.08 months, IQR 3.8-6.4) CAC was still
                             Medicine and Intermediate Care Unit, AOU
                                                                   er


                                                                                 present in 7 patients (1.4%); none of them required
                    Careggi, Florence; 2Department of Experimental and
                                                                                 increase of intensity medical care nor developed throm-
                    Clinical Medicine, University of Florence;
                                                              m



                                                                                 botic complication during the hospitalization. 3 (0.6 %)
                    Atherothrombotic Center, AOU Careggi, Florence, Italy
                                                                                 patients had thrombotic events after discharge; none of
                                                  om




                    Background and Aims: Covid associated coagulopathy           them had CAC at the follow up evaluation. Patients who
                    (CAC) in SarsCoV2 infected patients is a recognized          had developed thrombotic events during the hospitaliza-
                    marker of disease’s severity, however the usefulness of      tion still have high platelets level at follow up (244 vs
                                                                                 233 x 109/L; p=0.038). Prothrombin time (PT) was
                                               -c




                    its monitoring after hospital discharge is not defined and
                    poorly studied. We analyzed the prevalence of CAC and        more prolonged in both groups compared to controls.
                                                                                 Patients with more severe previous pneumonia and
                                       on




                    other hemostatic alterations in the follow up period of
                    patient with previous hospitalization for COVID related      thrombotic complications were more often on anticoag-
                    pneumonia to investigate their role as post discharge        ulant treatment and PT didn’t significantly differ
                                  N




                    thrombotic risk factors. Methods: This is a retrospec-       between mild and severe cases and between previous
                    tive, monocentric observational study. We collect data       thrombotic events or not grouping the patients for anti-
                    from electronic archives of Careggi Hospital, in             coagulant treatment. Conclusions: In our population the
                    Florence. We collect blood exams performed at hospital       presence of CAC at hospital admission was associated
                    admission and at the follow up evaluation, the intensity     with more severe illness and development of thrombotic
                    of medical care required and incidence of thrombotic         complication. Otherwise the presence of CAC during
                    events (deep vein thrombosis and pulmonary embolism)         the follow up seems have no value as a marker of previ-
                    during hospitalization and after discharge. CAC was          ous disease severity or thrombotic risk after discharge.
                    defined according to the presence of almost two of the       Our results also shows that patients who developed
                    following criteria: (1) decrease in platelet count (less     thrombotic complication during the hospitalization had
                    than 150 × 109/L); (2) increase in D-dimer (more than        higher platelets levels on admission and maintains this
                    two times the upper limit of normal); (3) >1 s prolonged     association at follow up. PT prolongation at follow up
                    prothrombin time or INR >1.2. Results: 430 patients          seems reflecting anticoagulant treatment instead of pre-
                    were identified, 249 man (57.9%) and 181 woman               vious disease severe or thrombotic risk. In conclusion
                    (42.1%), with median age of 65 years (IQR 55-74). 109        our results do not support the need of monitoring of
                    patients with an intensive or sub-intensive treatment        coagulation parameters after the discharge form the hos-
                    during hospitalization, 43 (7%) with no oxygen treat-        pital and shows that post discharge thrombotic risk was
                    ment during hospitalization, 30 patients (7%) had            very low.




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         Poster




                     PO002                                                          population of Tuscany (0.016), while for the FV Leiden
                                                                                    was 0.016, comparable to that observed in the tuscan pop-
                                                                                    ulation (0.020). Among a subgroup of n=324 patients, for
                     IDENTIFICATION OF RARE COAGULATION
                                                                                    which information concerning the clinical outcome was
                     VARIANTS AND SCREENING OF THROMBOPHILIC                        available, emerged that in patients who developed a
                     POLYMORPHISMS (F2 RS1799963, F5 RS6025)                        thromboembolic event (5.9%) there was a higher allelic
                     IN COVID-19 PATIENTS                                           frequency, but not statistically significant, of FVL poly-
                                                                                    morphism, compared with patients who did not develop
                     M. Giannini, E. Sticchi, T. Capezzuoli, R. De Cario,           such an event (0.026 vs. 0.018, p=0.519); no differences
                     S. Suraci, G. Cassioli, S. Neroni, A.A. Rogolino,              were observed for F2 G20210A polymorphism.
                     M. Berteotti, A.M. Gori, R. Marcucci, B. Giusti                Conclusions: The presence of common genetic factors of
                                                                                    hereditary thrombophilia does not seem to indicate a sig-
                     Dipartimento di Medicina Sperimentale e Clinica,               nificant contribution in modulating the risk of developing
                     Università degli Studi di Firenze; Atherothrombotic            thromboembolic complications in SARS-CoV-2 patients;
                     Diseases Center, AOU Careggi, Firenze, Italy                   on the other hand, NGS results show that genetic variabil-
                                                                                    ity, due to rare variants, might modulate clinical severity
                     Background and Aims: Severe SARS-CoV-2 infections              of COVID-19 disease in patients.
                     are characterized by perturbation of physiological coagu-
                     lation mechanisms. COVID-19 is associated with a high




                                                                                                            ly
                     incidence of thrombotic complications, such as venous          PO003
                     thromboembolism or arterial thrombotic events (myocar-




                                                                                                     on
                     dial infarction and cerebral events). An association
                     between thrombophilia and the most severe clinical             THE DIAGNOSTICS OF HEPARIN-INDUCED
                     course of COVID-19 has been suggested. Thrombophilia           THROMBOCYTOPENIA IN ITALY AND THE IMPACT




                                                                                               e
                     is a condition of altered haemostasis, characterized by        ON IT OF THE COMING TO LIGHT OF VITT
                     increased blood clotting, which predisposes to adverse
                     thrombotic events. This condition may be due to heredi-
                     tary factors, acquired changes or, in the prevalence of
                                                                                        us
                                                                                    E. Falcinelli1, R. Marcucci2, P. Gresele1
                                                                                    1Department   of Medicine and Surgery, Section of
                                                                                    al
                     cases, an association of genetic and acquired factors.
                     Among the hereditary thrombophilia factors, the most fre-      Internal and Cardiovascular Medicine, University of
                                                                                    Perugia, Italy; 2Center for Atherothrombotic disease,
                                                                            ci

                     quent causes are factor V Leiden polymorphism (rs6025)
                     in F5 gene and the G20210A polymorphism (rs1799963)            AOU Careggi, Department of Experimental and Clinical
                                                                     er


                     in F2 gene. Aim of the study was to identify genetic vari-     Medicine, University of Florence, Italy
                     ants and/or genetic profiles associated with severity of the
                                                                m



                     disease and thrombotic events susceptibility. Methods:         Background and Aims: Although rare, heparin-
                     Starting from these considerations, an NGS analysis was        induced thrombocytopenia (HIT) is a potentially omi-
                                                   om




                     conducted on a cohort of n=40 patients with COVID-19;          nous side effect of heparin treatment and must be
                     genetic analysis included a sequencing panel of 11 genes       promptly recognized to prevent serious consequences.
                     (PROC, PROS1, FGA, FGB, FGG, SERPINC1, F2, F5,                 The diagnosis is suspected clinically, by taking into
                                                                                    account several factors, among which the 4T pre test
                                                -c




                     F10, PLAT, PLG) known to be involved in the coagula-
                     tion process. Moreover, a genotyping analysis of rs6025        probability score, and requires laboratory confirmation
                                        on




                     and rs1799963 polymorphisms has been conducted                 by the detection of antibodies against PF4/heparin.
                     through Real Time PCR on the whole cohort of n=994             Tests include immunological assays, such as ELISAs
                     patients hospitalized at the AOU Careggi with COVID-           (enzyme linked immunosorbent assays) and others, as
                                   N




                     19. Results: As regards NGS analysis, 29 rare variants         well as a range of functional test exploring if circulating
                     (MAF≤1%) have been identified at the heterozygous state        antibodies can activate platelets. Most of these are
                     in 24 of the 40 patients studied: 7 missense variants (on      available only in specialized centers. In the wake of the
                     the F10, F2, PLAT, SERPINC1, F5 and FGB genes), 13             large-scale SARS-CoV-2 vaccine roll-out, a rare but
                     synonymous variants (on FGB, F2, PLAT, PLG, PROC               potentially fatal complication of adenoviral-vector vac-
                     and F5 genes), 4 variants concerning zone 3’-5’                cines, called vaccine-induced immune thrombotic
                     UTR/downstream (on F10, PLAT, PROC and F5 genes)               thrombocytopenia (VITT), has emerged with a vast
                     and 5 non-deep intronic variants (on F5, PROC, F10 and         negative impact on mainstream media. Similar to HIT,
                     PLG genes). In particular, there are five rare variants        also this novel catastrophic thrombotic syndrome
                     which were identified in two different patients each. A        requires the detection of high levels of anti-PF4 anti-
                     higher prevalence of rare missense variants with potential     bodies, either by ELISA (possibly with two or more dif-
                     pathogenic prediction in ICU or death patients (26.7%)         ferent assays) or by a functional assay. Our study aimed
                     was observed than in ordinary ward patients (8%).              to assess how VITT emergence has influenced the
                     Concerning common genetic thrombophilia, in the whole          request for these tests in an acute care setting of Italian
                     n=994 patients cohort, n=45 were heterozygous for the          hospitals. Methods: A 10-item self-administered ques-
                     rs1799963 polymorphism and n=31 were heterozygous              tionnaire was sent by email to all specialized laborato-
                     for rs6025 polymorphism. MAF for F2 G20210A was                ries performing anti-PF4 testing in Italy. Data collection
                     0.023, higher than that reported in the literature for the     included: methods for anti PFA/heparin identification,




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    request for the 4Tscore, total and median number of           P. Preti9, L. Sarti10, A. Trovati10, E. Lotti1,
                    requests per year and per month, rate of positive tests.      G. Palareti1, R. Marcucci2,11
                    A direct comparison between the period following the
                    emergence of VITT (from 01 March 2021 to 30 January           1Fondazione    Arianna Anticoagulazione, Bologna;
                                                                                  2SOD   Malattie Aterotrombotiche, Azienda Ospedaliero
                    2022) and the preceding 3 years (from 01 January 2018
                    to 31 December 2020) was made. Results: A total of 28         Universitaria-Careggi, Firenze; 3Dipartimento Di
                    laboratories (40% of the invited centers), from almost        Medicina e Chirurgia, Università Degli Studi
                    all regions of Italy, responded to the survey. Only 39%       Dell’Insubria, Varese; 4UO Medicina Generale
                    of respondents ask for the results of the clinical pre test   Ospedale SValentino, Montebelluna, TV; 5UO
                    probability score (4Tscore). The most commonly used           Medicina Interna ad Alta Intensità Azienda
                    laboratory assays for HIT antibodies are immunological        Ospedaliero-Universitaria        Careggi     Firenze;
                                                                                  6Dipartimento     di Medicina Interna, Ospedale
                    (68%), with only five laboratories using two different
                    immunological assays at the same time, while function-        Maggiore “Carlo Alberto Pizzardi”, Bologna;
                                                                                  7Dipartimento       di     Medicina      e    Scienza
                    al tests are performed only in a minority of laboratories
                    (32%). Among immunological tests the most used were           dell’Invecchiamento, Università GD’Annunzio, Chieti;
                                                                                  8U.O.CAngiologia, Ospedale Castelfranco Veneto,
                    ELISA (n=17) and chemiluminescence (n=10) followed
                    by a range of less used assays (n=3 latex test and n=2        Azienda ULSS 8-Regione Veneto, Castelfranco Veneto;
                                                                                  9Thrombosis and Haemostasis Centre, Department of
                    rapid assay). The main commercial sources of reagents
                    are Immucor (46%) and Werfen (43%), followed by               Internal Medicine, IRCCS SMatteo Foundation,




                                                                                                           ly
                    Stago (14%). Total number of tests/year was signifi-          University of Pavia, Pavia; 10UOC Medicina Interna
                                                                                  d’Urgenza e Area Critica, Ospedale di Baggiovara,




                                                                                                   on
                    cantly increased comparing the post VITT period to the
                    preceding 3 years (n=138, 95%CI: 31.3-302 vs 58.5,            Azienda Ospedaliero Universitaria Modena;
                                                                                  11Dipartimento di Medicina Sperimentale e Clinica,
                    95%CI: 9.4-115; +138% p<0.05) (Figure 1A). The
                    number of tests/month was also increased from n=6.04          Università degli Studi di Firenze, Italy




                                                                                             e
                    (95%CI: 1.1-10.5) to n=10.41 (95%CI: 2.3-22.8)
                    (+66%) (Figure 1B). No difference in the incidence rate
                    of positive tests was observed in the two periods (8%,
                    95%CI: 3.8-11 vs 11%, 95% CI: 5-20;p=ns).
                                                                                      us
                                                                                  Background and Aim: COVID-19 infection causes
                                                                                  acute respiratory pathology insufficiency with severe
                                                                                  interstitial pneumonia and extra-pulmonary complica-
                                                                                  al
                    Conclusions: The results of this survey, which included       tions; in particular, it may predispose to thromboem-
                    responses from 28 laboratories representing 14 regions        bolic disease. The reported incidence of thromboem-
                                                                          ci

                    along Italy, showed important differences in laboratory       bolic complications varies from 5% to 30% of cases.
                    practice for HIT diagnosis and an important impact of         Methods: We conducted a multicentre, Italian, retro-
                                                                    er


                    the coming to light of the VITT complication in Italy.        spective, observational study on COVID-19 patients
                    The use of HIT assays markedly increased after VITT           admitted to ordinary wards, to describe clinical char-
                                                              m



                    identification, although this complication is very rare.      acteristic of patients at admission, bleeding and
                    The major cause behind this increase is likely related to     thrombotic events occurring during hospital stay.
                                                  om




                    the hyper vigilance among emergency physicians raised         Results: 1135 patients hospitalized were included in
                    by this unprecedented complication of vaccination.            the START-COVID-19 Register, 1091 patients hospi-
                                                                                  talized in ordinary wards were included in the study,
                                               -c




                                                                                  653 males (59.9%), median age 71 years (IQR 59-82
                                                                                  years). During the observation, 2 (0.2%) patients had
                                       on




                                                                                  acute coronary syndrome episodes, 1 patients (0.1%)
                                                                                  had a stroke; no other arterial thrombotic events were
                                                                                  recorded. Fifty-nine patients had symptomatic VTE
                                  N




                                                                                  (5.4%), 18 (30.5%) patients had deep vein thrombosis
                                                                                  (DVT), 39 (66.1%) patients had pulmonary embolism
                                                                                  (PE), and 2 (3.4%) patients had DVT + PE. Among
                                                                                  patients with DVT, 8 (44.4%) were isolated distal
                    Figure 1.                                                     DVT, and 2 cases were jugular thrombosis. Among
                                                                                  patients with PE, 7 (17.9%) events were limited to
                                                                                  sub-segmental arteries. No fatal PE were recorded.
                    PO004                                                         Major bleedings occurred in 9 (1.2%) patients, and
                                                                                  clinically relevant non- major bleeding in 9 (1.2%)
                                                                                  patients. All bleedings occurred among patients treat-
                    THROMBOEMBOLIC COMPLICATIONS IN                               ed with antithrombotic drugs receiving thrombopro-
                    PATIENTS WITH COVID-19 HOSPITALIZED IN                        phylaxis, more frequently when treated with sub-ther-
                    ITALIAN ORDINARY WARDS: DATA FROM THE                         apeutic/therapeutic dosages. Conclusions: We con-
                    MULTICENTRE OBSERVATIONAL START-COVID                         firmed that patients admitted to ordinary wards for
                    REGISTER                                                      COVID-19 infection are at high risk for thromboem-
                                                                                  bolic events. VTE recorded among these patients are
                    E. Antonucci1, D. Poli2, W. Ageno3, S. Galliazzo4,            mainly isolated PE, suggesting a peculiar characteris-
                    A. Milia5, A Pizzini6, M. Di Nisio7, B. Zalunardo8,           tic of VTE in these patients.




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         Poster




                     PO005                                                        ELISA and the chemiluminescence test; instead the HIPA
                                                                                  test confirmed positivity (4/4 buffer, 4/4 PF4 10 µg/mL,
                                                                                  and 4/4 heparin 0.2 U/mL). The patient was re-evaluated
                     VITT OR NOT VITT? 61-YEARS WOMAN WITH                        after further 30 days and the results of test were negative
                     THROMBOCYTOPENIA AFTER SOMMINISTRATION                       both at immunological (ELISA test and chemilumines-
                     OF AD26.COV2.S VACCINE                                       cence test) and functional tests. Conclusions: This is one
                                                                                  of the rare cases of discordance between the immunoas-
                     A. Bertelli, A.A. Rogolino, F. Cesari, B. Giusti,            say and the functional test. It occurs in HIT in about 0.2%
                     E. Sticchi, B. Bandinelli, E. Lotti, A.M. Gori,              of cases and can be ascribed to the presence of antibodies
                     R. Marcucci                                                  directed against other than PF4/Heparin complex such as
                                                                                  Nap2- and IL8-Heparin complexes. In fact, these com-
                     Department of Experimental and Clinical Medicine,            plexes were not recognized by the classical immunoas-
                     University of Florence; Atherothrombotic Diseases            say anti-PF4, which are specific for the PF4/Heparin
                     Center, AOU Careggi, Florence, Italy                         polyanion complexes. Therefore, the discordance
                                                                                  between immunological and functional tests may be
                     Background: Vaccine-induced immune thrombotic                explained by the presence of these antibodies in our
                     thrombocytopenia (VITT) is a rare but severe immuno-         patient, in whom the timely pharmacological interven-
                     logical reaction to the non-replicable adenoviral vector-    tion likely stopped the progression of the disease.
                     based COVID-19 vaccines (AstraZeneca vaccine and




                                                                                                          ly
                     Johnson & Johnson). VITT is caused by antibodies that
                     recognize platelet factor 4 (PF4) bound to platelets.




                                                                                                   on
                                                                                  PO006
                     These antibodies are immunoglobulin G (IgG) molecules
                     that activate platelets via low-affinity platelet FcγIIa
                     receptor. Extreme activation of platelet and the coagula-    INDIVIDUAL SUSCEPTIBILITY TO SARS-COV-2




                                                                                             e
                     tion system leads to clinically significant thromboembol-    INFECTION: ROLE OF THE HOST’S VIRUS ENTRY
                     ic complications. Case Report: We describe the case of
                     a 61-years woman, who presented in June 2021 to the
                     emergency department for bruising and conjunctival
                                                                                      us
                                                                                  MACHINERY GENETIC PROFILES

                                                                                  R. De Cario1, E. Sticchi1, A. Kura1, T. Capezzuoli1,
                     hemorrhagic manifestations. Patient had been adminis-        M. Giannini1, G. Cassioli1, G. Barbieri1, S. Suraci1,
                                                                                  al
                     tered Ad26.COV2.S vaccine (J&J) 20 days earlier. Her         A.M. Gori1, M. Berteotti2, A.A. Rogolino1,
                                                                                  R. Marcucci1, B. Giusti1
                                                                          ci

                     medical history showed hypothyroidism and hyperten-
                     sion. Blood tests showed thrombocytopenia with platelet
                                                                    er


                                                                                  1Department of Experimental and Clinical Medicine,
                     count of 38x109/L. In the days following hospitalization,
                     the patient was administered with corticosteroids and        University of Florence; Atherothrombotic Diseases
                                                              m



                     intravenous immunoglobulin (IVIG) (400 mg/kg). Then          Center, AOU Careggi, Florence, Italy; 2Cardiomyopathy
                     platelet count dropped to 23x109/L, and the patient was      Unit, Cardiothoracic and Vascular Department and
                                                  om




                     admitted for further investigations. The temporal rela-      Cardiomyopathy Unit, Careggi University Hospital,
                     tionship between the onset of symptoms and vaccine           Florence, Italy
                     administration raised the suspicion of VITT.
                                                                                  Background: Since the 2019 SARS-CoV-2 pandemic
                                               -c




                     Subsequently patient was administered with corticos-
                     teroids and intravenous immunoglobulin (IVIG) (400           outbreak, hundreds of studies reported evidence of a host
                                        on




                     mg/kg). Samples for platelet factor 4 (PF4) antibody         genetic background’s role, alongside age, comorbidities,
                     analysis were sent to the laboratory of the                  and gender, in influencing both susceptibility to virus
                     Atherothrombotic Centre of Florence to confirm the           infection and severity of the clinical pictures. Genes-reg-
                                   N




                     diagnosis. The titre of antibodies anti-PF4 was assessed     ulated biological pathways of COronaVIrus Disease
                     by using two enzyme-linked immunosorbent assay               2019 (COVID-19) suggested to be mainly implicated in
                     (ELISA), PF4 IgG test, Immucor-Lifecodes and                 the different clinical outcomes are represented by virus
                     Zymutest HIA IgG, Hyphen BioMed-, and a chemilumi-           entry, immunity, and inflammation. Concerning the first,
                     nescent assay (Acustar Werfen). Patient’s sample was         several factors are involved both at the cell surface and
                     negative with all immunoassays: 0.125 optical density        intracellular level. Several genetic variants
                     (OD) for Immucor, 0.109 OD for Zymutest and 0.10             (mutations/polymorphisms) were found to be associated
                     U/mL for Acustar. To evaluate the capability of anti-PF4     with the severity of the disease in a detrimental or bene-
                     antibodies in inducing platelet activation, a functional     ficial way, their identification potentially representing
                     test (HIPA Test) was also performed. Unexpectedly, at        useful prognostic/therapeutic tool. In this study, genetic
                     the HIPA test we observed a strong capacity of patient’s     characterization of 40 Covid19 patients referred to the
                     serum in inducing platelet activation both in the presence   Advanced         Molecular       Genetics       Laboratory,
                     of only buffer and in the presence of PF4 (10 µg/mL).        Atherothrombotic Diseases Center, Careggi Hospital-
                     The discordance between immunoassays and the HIPA            University of Florence, was made by Next Generation
                     test suggested a further evaluation at 30 days. In the       Sequencing (NGS) to identify genetic profiles potentially
                     meantime, the platelet count increased to 166x109/L and      representing prognostic factors modulating the suscepti-
                     there was no evidence of thrombosis. The test was            bility to virus infection. Methods: NGS was performed
                     repeated at 30 days and confirmed the negativity of both     by Illumina MiSeq and Haloplex HS protocol targeting 7




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 85



                                                                           XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    virus entry-related genes (ACE2, TMPRSS2, CTSL,                 of patients develop, instead, a serious disease that can
                    CTSB, HSPA5, IL6, FURIN). Results: 18 heterozygous              evolve into acute respiratory distress syndrome (ARDS),
                    rare variants [Minor Allele Frequency (MAF)<0.01] were          shock, macro and microvascular thrombosis, multiorgan
                    identified in 16/40 patients involving all 7 genes included     dysfunction with poor prognosis. It has been shown that
                    in the panel. Potentially functional mutations (10/18)          patients with COVID-19 exhibit a hypercoagulable pro-
                    involved ACE2, HSPA5, CTSL with TMPRSS2 being the               file characterized by an increase in D-dimer, fibrinogen,
                    most affected gene, in accordance with literature data          factor VIII and von Willebrand factor (vWF) plasma lev-
                    which identified this gene as promising therapeutic target.     els and an increased whole blood coagulation capability
                    Some of those rare variants were interpreted as protective      as        identified        by      viscoelastic       tests
                    due to their potential role in diminishing the activity of      (thromboelastometry/graphy). The first aim of our study
                    the virus-entry machinery (i.e. ACE2), others may instead       was to evaluate the possible presence of a hypofibrinoly-
                    increase the susceptibility to infection (i.e. TMPRSS2).        sis condition in subjects with acute COVID-19 pneumo-
                    With regards to the disease progression in terms of             nia. Secondly, we evaluated the possible role of these
                    death/hospitalization in intensive care unit (ICU, 15 sub-      alterations in determining a state of hypercoagulability
                    jects, with 3 variants) or early discharge (25 subjects, car-   which could contribute to increase the thrombotic risk in
                    rying 7 variants), no statistically significant difference      these patients. Methods: We performed a case-control
                    was observed among the 2 rare variants carriers’ groups         study that included 63 patients, admitted to the Internal
                    (20% vs 28%, showing a slight tendency towards a pro-           Medicine Ward of University of Padova between
                    tective effect of rare variants). About the polymorphic         November 6th, 2020 and April 26th, 2021 with a diagno-




                                                                                                             ly
                    burden, the most affected genes were FURIN, CTSB, and           sis of SARS-CoV-2 infection, as defined by WHO. In




                                                                                                     on
                    TMPRSS2, with an average number of variants of 1.4, 4.1         each patient we studied the coagulation profile with par-
                    and 3.3 per patient, respectively. Considering the total        ticular reference to the fibrinolytic system, by means of
                    number of polymorphisms per gene, the only slight dif-          traditional coagulation tests and by rotational thromboe-
                    ference among the abovementioned subgroups was                  lastometry conducted on whole blood using the




                                                                                               e
                    observed regarding the FURIN gene as 37 polymor-                ROTEM® method. The data obtained in this group of
                    phisms affected the early discharge group, while 18 (67%
                    vs 83%) were carried by the death/ICU group.
                    Conclusions: Our data suggest how the identified genetic
                                                                                        us
                                                                                    patients (cases) were compared with those obtained in a
                                                                                    group of healthy subjects (controls) sex and age matched
                                                                                    with cases. Results: In patients with COVID-19 related
                                                                                    al
                    variants involving the host’s virus entry machinery are         pneumonia, we found a significant increase in ferritin,
                    not likely to exert a significant effect in modulating the      CRP, procalcitonin and IL-6 plasma levels. The tradition-
                                                                            ci

                    severity of the COVID-19 progression when considering           al coagulative profile, characterized by a significant
                    the prognosis parameters of death, ICU hospitalization          increase of D-dimer, fibrinogen, FVIII and vWF, demon-
                                                                     er


                    and early discharge from the hospital. According to our         strated a state of hypercoagulability. We also found a sig-
                    data, the contribution of other factors (genetic or not), i.e   nificant increase of both tPA and PAI plasma levels in
                                                               m



                    comorbidities and individual thrombophilic state, are           cases than controls. Finally, according to thromboelasto-
                    more likely to contribute to the disease severity and the       graphic tests we found a statistically significant reduction
                                                   om




                    potential complications at different levels (coagulation        of Clot Formation Time (CFT) and Maximum Lysis
                    processes, cardiac or neurological involvement, exten-          (ML), both in the INTEM and EXTEM tests, and a sta-
                    sion of the pulmonary injury) along with inflammation,          tistically significant increase of Maximum Clot Firmness
                                                -c




                    and immunity process, that may be, at least in part, influ-     (MCF) in INTEM, EXTEM and FIBTEM assays.
                    enced by alterations at the virus-entry level.                  Conclusions: The results of our study allowed us to con-
                                        on




                                                                                    firm that patients with acute COVID-19 related infection
                                                                                    have a state of hypercoagulability. Furthermore, the
                    PO007                                                           study of the fibrinolytic parameters revealed a picture of
                                   N




                                                                                    hypofibrinolysis. Larger studies are needed to identify a
                                                                                    possible correlation between our findings and the devel-
                    CLINICAL IMPLICATIONS OF FIBRINOLYTIC
                                                                                    opment of thrombotic complications.
                    SYSTEM ALTERATIONS IN PATIENTS WITH
                    COVID-19 RELATED PNEUMONIA
                                                                                    PO008
                    L. Spiezia, M. Isola, C. Simion, E. Campello,
                    F. Dalla Valle, P. Simioni
                                                                                    POST-DISCHARGE THROMBOPROHYLAXIS IN
                    Thrombotic and Hemorrhagic Diseases Unit, Padova                PATIENTS WITH COVID-19: A SINGLE-CENTER
                    University Hospital, Padova, Italy                              EXPERIENCE

                    Background and Aims: SARS-CoV-2 (Severe Acute                   C. Simion, E. Campello, D. Tormene, L. Spiezia,
                    Respiratory Syndrome Coronavirus-2) infection that              F. Dalla Valle, P. Simioni
                    causes COVID-19 (Coronavirus Disease-2019) is a glob-
                    al pandemic. The clinical manifestations of SARS-CoV-           General Medicine and Thrombotic and Hemorrhagic
                    2 infection are extremely varied, most patients (80%)           Diseases Unit, Department of Medicine – DIMED,
                    develop a mild upper respiratory tract infection, but 15%       University-Hospital of Padova, Italy




                                                                [Hematology Reports 2021; 13:s3]                                                   [page 85]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 86



         Poster




                     Background and Aims: COVID-19 is associated with             prophylaxis up to 14-20 days may be more effective in
                     an increased risk of venous thromboembolism (VTE).           preventing any VTE. Post-discharge mortality was non-
                     However, data regarding rates of VTE after discharge         significantly higher in patients without extended throm-
                     for patients hospitalized for COVID-19 are limited and       boprophylaxis.
                     extended out-of-hospital thromboprophylaxis is not
                     routinely recommended because the risk-benefit               Table 1.
                     remains uncertain. Methods: We enrolled all consecu-
                     tive adults admitted to our General Internal Medicine
                     Unit for confirmed COVID-19 between November
                     2020 and May 2021. We collected demographics,
                     comorbidities, laboratory findings, risk factors for VTE
                     and medications, including in-hospital and post-dis-
                     charge thromboprophylaxis. Any VTE, bleeding event,
                     re-hospitalization and all causes of mortality within 30
                     days after discharge were considered as outcomes.
                     Results: Out of 388 enrolled adults (male 51.0%, medi-
                     an age 77 years), 70 (18%) were excluded because
                     already receiving therapeutic anticoagulation before the
                                                                                  PO009
                     hospitalization, mostly (72.9%) for atrial fibrillation




                                                                                                         ly
                     (AF). Forty-five subjects (11.6%) died during hospital-
                     ization, mainly because of respiratory or multiorgan         HYPERINFLAMMATORY STATE AND DEATH IN




                                                                                                  on
                     failure and septic shock. Eight patients (2.9%) did not      PATIENTS AFFECTED BY COVID-19: ROLE OF
                     receive thromboprophylaxis during the hospitalization        VACCINES
                     (2 died within the first 24 hours for respiratory failure,




                                                                                             e
                     6 had a major bleeding at admission). During hospital-       L. Marchini, M.C. Vedovati, C. Becattini
                     ization, full anticoagulation treatment was required in
                     26.0%: 19 cases (7.0%) for AF onset and 49 (17.9%) for
                     VTE occurrence, mainly distal or catheter-related deep
                                                                                      us
                                                                                  Internal, Vascular and Emergency Medicine – Stroke
                                                                                  Unit, University of Perugia, Perugia, Italy
                                                                                  al
                     venous thrombosis (DVT); three patients developed
                     both AF and VTE. Out of 202 (74.0%) patients dis-            Background: Systemic hyperinflammation in patients
                                                                          ci

                     charged without full anticoagulation, 25 (12.4%) did         with COVID-19 is associated with a higher probability
                     not received thromboprophylaxis, while 177 (87.6%)           of severe disease and poor prognosis. The COVID-19
                                                                    er


                     received extended prophylaxis with enoxaparin 4000           vaccines are expected to be associated with a lower risk
                     IU subcutaneous daily (6000 IU if obese). Particularly,      of moderate-severe disease. However, data on the effects
                                                              m



                     25 patients (12.4%) received less than 7 days of prophy-     of vaccination on the inflammatory state and adverse
                     laxis, 99 (49%) up to 7-10 days, 4 (2.0%) up to 15-21        outcomes in patients affected by COVID-19 are lacking.
                                                  om




                     days, 1 (0.5%) 30 days because of recent hip fracture;       Aims: To evaluate the association of hyperinflammation
                     for 48 subjects (23.7%) the duration of thromboprophy-       state and adverse outcome among vaccinated and unvac-
                     laxis was not specified (until complete                      cinated patients for SARS-CoV-2 infection. Methods:
                                               -c




                     mobilization/negativization/full recovery). Among the        We performed an observational study on COVID-19
                     177 patients who received post-discharge prophylaxis,        patients admitted to a non-ICU ward at Perugia Hospital
                                        on




                     2 (1.1%) VTE events occurred during the follow-up: a         from August to December 2021. The inclusion criteria
                     proximal DVT of the leg occurred in one patient (male,       were: age ≥18 years, hospitalization due to respiratory
                     56 years, heterozygous FV Leiden, with a past history        failure, and SARS-CoV-2 infection confirmed by
                                   N




                     of distal DVT, who had severe COVID-19) soon after           nasopharyngeal RT-PCR swab. A patient was defined as
                     stopping a 7 day-prophylaxis; a 71 years old female          vaccinated after a full cycle. Study outcomes were all-
                     patient was re-admitted 42 days after for a stroke due to    cause-death or symptomatic venous thromboembolism
                     paradoxical embolism from proximal DVT; she had a            (VTE) and hyperinflammation state (defined as at least
                     history of previous stroke and received a 7 day-throm-       four values among CRP, LDH, ferritin, CPK, D-dimer
                     boprophylaxis after discharge, but did not recover full      above the threshold). Results: Overall, 182 patients
                     mobilization. Five (2.8%) patients were re-hospitalized      were included (mean age 68 years, range 18-98). All-
                     within 30 days, not relating to VTE or bleeding; 4           cause death occurred in 16 patients (8.8%). Vaccinated
                     patients (2.3%) died within 16 days (1 multiorgan fail-      patients were older (76 vs 61 years); they had a higher
                     ure, 1 worsening of general condition, 2 unknown rea-        rate of comorbidities and a lower rate of NIV/HFNC
                     son) and 1 (0.6%, cancer) 32 days post discharge.            requirements than unvaccinated patients. After age
                     Among the 25 patients discharged without prophylaxis,        adjustment, the hyperinflammation state was signifi-
                     neither VTE nor bleeding were recorded, there were no        cantly more frequent in unvaccinated compared to vac-
                     re-hospitalization but 2 patients (8.0%) died within 15      cinated patients (65 vs 37%, p=0.004). Lack of vaccina-
                     days (p vs extended prophylaxis 0.12) (Table 1).             tion was an independent predictor of in-hospital all-
                     Conclusions: According to our findings, extended post-       cause-death (HR 2.71, 95% CI 1.05-7.00, p=0.040) and
                     discharge thromboprophylaxis up to 7 days appears to         all-cause-death or symptomatic VTE (HR 3.10, 95% CI
                     be safe in patients with COVID-19. A longer course of        1.31-7.35, p=0.010). The risk of symptomatic VTE was




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 87



                                                                       XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    not significantly higher in unvaccinated compared to       35 patients without prophylaxis : RR 1.32 (95% CI 0.28
                    vaccinated patients (5 vs 1%, HR 5.66, 95% CI 0.63-        to 6.2). Twenty-seven patients in the prophylaxis group
                    50.85). Conclusions: Covid-19 vaccination is associat-     died versus 3 in the therapy group and 6 in the non-pro-
                    ed with a lower hyperinflammation state and lower risk     phylaxis group: RR 1.97 (95%ci 0.62 to 6.2) and 2.44
                    of death compared to lack of vaccination. These findings   (0.65 to 9.1) respectively. Major bleeding occurred in
                    should be confirmed in a larger population.                11 % of the patients receiving therapeutic-dose antico-
                                                                               agulation and in 5 % of those receiving thromboprophy-
                                                                               laxis. 81 patients were transferred to the ICU. Of these
                                                                               30 developed a thrombotic event: 24 thrombotic events
                                                                               [1 proximal DVT ] occurred in the group of 64 patients
                                                                               on prophylactic heparin, versus 4 thrombotic events [1
                                                                               proximal DVT] in the group of 8 patients on heparin at
                                                                               therapeutic or sub-therapeutic doses: RR 0.82 (95% CI
                                                                               0.34 to 1.95). Two thrombotic events [distal DVT]
                                                                               occurred in the 9 patients without prophylaxis : RR 0.54
                                                                               (95% CI 0.12 to 2.4). Fifteen patients in the prophylaxis
                                                                               group died versus 1 in the therapy group and 4 in the
                                                                               non-prophylaxis group: RR 1.71 (95%ci 0.25 to11.5)
                                                                               and 2.77(0.37 to 21) respectively. Conclusions: The




                                                                                                       ly
                                                                               risk of developing thrombotic events in patients with
                                                                               Covid 19 admitted to an internal medicine ward in




                                                                                                on
                    Figure 1.                                                  which heparin prophylaxis is performed does not
                                                                               appear significantly increased. There appears to be an
                                                                               increased risk of death in patients receiving prophylac-




                                                                                          e
                    PO010                                                      tic heparin doses compared to those receiving therapeu-

                    INCIDENCE OF VENOUS THROMBOEMBOLISM IN
                                                                                   us
                                                                               tic or sub-therapeutic doses. This employer is less evi-
                                                                               dent in those transferred to the ICU. An adequate
                                                                               assessment of the patient’s haemorrhagic risk will be
                    HOSPITALIZED PATIENTS WITH COVID-19 AND
                                                                               al
                                                                               useful in the most appropriate treatment choice.
                    ANTICOAGULATION WITH HEPARIN.
                                                                       ci

                    A SINGLE-CENTER COHORT STUDY
                                                                               PO011
                                                                 er


                    D. Tormene, E. Bicego, N. Perin, E. Campello,
                    M. Sartori, C. Simion, P. Simioni
                                                                               INFIAMMAZIONE VASCOLARE CORONARICA
                                                            m




                    Thrombotic and Haemorrhagic Diseases Unit,                 STIMATA MEDIANTE L’ATTENUAZIONE
                                                om




                    Department of Medicine, University of Padua Medical        PERI-VASCOLARE CORONARICA ALLA
                    School, Padua, Italy                                       TOMOGRAFIA COMPUTERIZZATA DEL TORACE
                                                                               NEI PAZIENTI RICOVERATI PER COVID-19
                    Background and Aims: Coagulopatthy and venous
                                             -c




                                                                               E MORTALITÀ INTRAOSPEDALIERA.
                    thromboembolism are common findings in patients
                    with Coronavirus disease. The aim of the study was to      D. Tuttolomondo1-2, C. Martini1-3, M. De Filippo1-3,
                                      on




                    investigate the incidence of objectively confirmed         N. Gaibazzi2
                    thrombotic events (both venous and arterial) in hospi-
                                 N




                    talized patients with COVID-19. Methods: Single-cen-       1Università degli studi di Parma; 2Azienda Ospedaliero-
                    ter cohort study of 284 hospitalized patients with         Universitaria Parma, UOC Cardiologia, Parma, Italia;
                    COVID-19. Results: Two hundred and eighty-four             3Azienda Ospedaliero-Universitaria Parma, UOC
                    patients were admitted to our internal medicine ward:      Radiologia, Parma, Italy
                    136 (48%) were male and the mean age was 77 years.
                    One hundred thirteen patients (40%) had cardiovascular     Background and Aims: SARS-CoV-2 acute respiratory
                    disease, 48 (17%) metabolic syndrome, 20 (7%) active       disease (COVID-19) can present with different clinical
                    cancer and 53 (19%) previous cancer. Prophylactic          scenarios ranging from flu-like symptoms to respiratory
                    doses of heparin were used in 202 patients, sub-thera-     distress syndrome. Up to 45.3% of patients hospitalized
                    peutic or therapeutic doses in 47 and in 35 no heparin     for COVID-19 have documented myocardial damage
                    prophylaxis was performed. Fifteen thrombotic events       (increased cardiac troponin); this determines a worsening
                    [6 DVT (4 distal), 4 myocardial infarcts, 5 cerebral       of the prognosis with an increase of in-hospital mortality.
                    ischemic events] occurred in the group of patients on      Cardiac involvement in these patients may be multifacto-
                    prophylactic heparin, versus three thrombotic events [1    rial but the prevailing hypothesis seems to be the throm-
                    distal DVT, 2 myocardial infarcts] in the group of         botic destabilization of the atherosclerotic plaque sec-
                    patients on heparin at therapeutic or sub-therapeutic      ondary to the “cytokine storm”. In this scenario, the esti-
                    doses: RR 1.16 (95% CI 0.32 to 4.2). Three thrombotic      mate of vascular inflammation should be contextualized
                    events [1TVP and 2 myocardial infarcts] occurred in the    by means of coronary peri-vascular attenuation (PVAT)




                                                             [Hematology Reports 2021; 13:s3]                                                [page 87]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 88



         Poster




                     with chest computed tomography (CT). This parameter           concerns developed regarding an increased risk of
                     can be obtained ex-post (offline) and does not require        thrombosis associated with thrombocytopenia: a new
                     further exams or additional costs. The prognostic power       syndrome of vaccine-induced immune thrombotic
                     of PVAT is known but has not yet been tested in COVID-        thrombocytopenia (VITT) has emerged as a rare side-
                     19 patients. Methods: Single-center retrospective study       effect of vaccination against COVID-19. Reports
                     conducted on 800 patients hospitalized for COVID-19           described a small number of cerebral venous thrombosis
                     from 01.03.2020 to 30.04.2020 and from 01.03.2021 to          among tens of millions of Vaxzevria vaccinated individ-
                     30.04.2021 who performed chest CT according to clini-         uals. In january 2022 the U.S. Vaccine Adverse Events
                     cal judgment. The objective of the study is to evaluate the   Reporting System (VAERS) identified 54 cases of
                     predictive capacity of PVAT respect to the primary end-       thrombosis with thrombocytopenia among over 14 mil-
                     point of all-cause-death and the composite end-point of:      lion recipients of Jcovden vaccine. Of these, one third
                     cardiovascular death, acute myocardial infarction,            had cerebral venous sinus thrombosis. The remaining
                     stroke/transient ischemic attack, major cardiac arrhyth-      cases have affected the splanchnic system, heart, lungs
                     mias, non-invasive ventilation, invasive ventilation and      or limbs. In Italy, 2 anaphylaxis, 1 acute or subacute
                     hospitalization in intensive care units. Results: 765         neuropathy (Guillain-Barrè syndrome), and 1 atypical
                     patients out of 850 patients were enrolled (study in          intracranial thrombotic event associated with thrombo-
                     progress) and PVAT analysis on chest CT was performed         cytopenia (VITT) can be counted per million doses
                     in 643 cases. A preliminary statistical analysis was per-     administered. With a prevalence of young women. Case
                     formed regarding the patients who had hospitalized for        Reports: We describe 11 fatal cases of suspected VITT,




                                                                                                           ly
                     COVID-19 from 01.03.2020 to 30.04.2020 (399                   7 females and 4 males aged 18-74, occurred in northern




                                                                                                   on
                     patients). PVAT mean in dead patients (n=175) calculated      Italy between february and july 2021. We studied all the
                     on the anterior interventricular artery (IVA) is -68.89 UH    cases on assignment from the Public Prosecutor Office
                     and on the right coronary artery (RCA) is -69.9 UH; in        at the Court. Cases were studied at autopsy, and relation-
                     surviving patients (n=224) the mean PVAT was -73.75           ship to the vaccine was considered according to the fol-




                                                                                              e
                     UH (IVA) and -72.66 (RCA) both p <0.01. Conclusions:          lowing criteria: age, sex, vaccine given, interval, throm-
                     Preliminary results show that the degree of coronary
                     inflammation on chest CT in hospitalized COVID-19
                     patients could be a useful parameter in prognostic strati-
                                                                                       us
                                                                                   bosis, thrombocytopenia, PF4 antibody testing. For
                                                                                   every case, 4 evaluation steps (Eligibility, Checklist,
                                                                                   Algorithm, Classification) according to the WHO
                                                                                   al
                     fication.                                                     Causality Assessment recommendation were applied.
                                                                                   Four autopsies (2 Vaxzevria and 2 Comirnaty vaccines)
                                                                           ci

                                                                                   revealed myocardial infarctions, classified as “indeter-
                     PO012                                                         minate”. Five autopsies (all Comirnaty vaccines)
                                                                      er


                                                                                   revealed two myocardial infarctions, one venous throm-
                                                                                   boembolism, one subarachnoideal hemorrage, one
                                                                 m



                     DEATHS AFTER ANTI SARS-COV-2 (COVID-19)
                                                                                   necrotizing pancreatitis, classified as “inconsistent with
                     VACCINE ADMINISTRATION IN ITALY:                              causal association to immunization”. Two autopsies of
                                                   om




                     AUTOPSY REPORT OF ELEVEN CASES                                young women (2 Vaxzevria vaccines) revealed cerebral
                                                                                   hemorrhage associated with thrombosis of the dural
                     C. Franzetti1, L. Tajana1, F. Piovella2                       venous sinuses and presence of anti-PF4 antibodies,
                                                -c




                     1Unità
                                                                                   classified as “consistent with causal association to
                            di Medicina Legale e Scienze Forensi                   immunization, vaccine product-related reaction as per
                                        on




                     “AFornari”, Dipartimento di Sanità Pubblica, Medicina         published literature”. Conclusions: Our results confirm
                     Sperimentale e Forense, Università di Pavia, Pavia;           the likely association of VITT and Vaxzevria vaccine in
                     2Angioematologia, Casa di Cura Lecco “Beato Luigi
                                                                                   two cases. In april, 2021 AIFA recommended a preferen-
                                   N




                     Talamoni”, Lecco, Italy                                       tial use of Vaxzevria and Jcovden in subjects older than
                                                                                   60. On may 5, 2022 FDA has limited the use of the
                     Background: As of may 12, 2022, 516.922.683 acute             Jcovden vaccine to individuals 18 years of age and older.
                     respiratory infections and 6.259.495 deaths from coron-       The trend of adverse reactions to vaccination against
                     avirus SARS-CoV-2 (COVID-19) have been reported               COVID-19 has been judged as acceptable from the clin-
                     worldwide. Venous thrombotic complications have been          ical and epidemiological point of view and the benefits
                     associated with up to 60% of these deaths. As a conse-        of the vaccine continue to outweigh the risks.
                     quence, we witnessed unprecedented rapid clinical
                     development of vaccines. As of may 10, 2022,
                     11.655.356.423 doses have been delivered. Four vac-           PO013
                     cines have been approved in Italy. Two are vaccines
                     based on incapsulated mRNA in lipid nanoparticles
                     (COMIRNATY, Pfizer/BioNTech and SPIKEVAX,                     LONG TERM FOLLOW-UP OF A MULTICENTRE
                     Moderna), two use a non-replicating recombinant chim-         COHORT OF COVID-19 PATIENTS WITH
                     panzee adenovirus to deliver genetic material                 PULMONARY EMBOLISM: ANTICOAGULATION
                     (VAXZEVRIA, Oxford–AstraZeneca and JCOVDEN,                   MANAGEMENT AND OUTCOMES
                     Johnson & Johnson/Janssen). Vaxzevria vaccine cam-
                     paign started in U.K. on January 4, 2021. In march 2021,      L. Filippi1, M. Milan2, G. Turcato1, S. Barbar3,




         [page 88]                                             [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 89



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    D. Tonello1, D. Milazzo1, A. Zaboli4, L. Panepinto5,        group (p=NS); no difference was also shown in risk of
                    E. Miozzo6, S. Cuppini2, M. Marchetti1                      VTE relapses (0% vs 4.1%, p=ns), major bleeding
                                                                                (4.5% vs 4.1%, p=NS) and in composite outcome (9.1%
                    1Internal Medicine Alto Vicentino Hospital Santorso;        vs 11%, p=ns). No difference was found between the
                    2Internal Medicine Rovigo Hospital “Santa Maria della       two treatment groups for the composite outcome at the
                    Misericordia”; 3Internal Medicine Cittadella Hospital;      Kaplan-Meier analysis (Log Rank Test p=0.387).
                    4Emergency department Merano Hospital “Franz
                                                                                Conclusions: In our retrospective multi-center cohort
                    Tappeiner”; 5Internal Medicine University of Verona;        prolongation of duration of anticoagulation seems not
                    6Internal Medicine University of Padua, Italy
                                                                                to affect risk of VTE recurrences, deaths and bleeding
                                                                                after a PE related to COVID-19.
                    Background and Aims: Pulmonary embolism (PE) is a
                    frequent complication in COVID-19 hospitalized
                    patients. Inflammatory storm and endothelial dysfunc-       PO014
                    tion due to the virus seem to be the two major risk fac-
                    tors for PE. Consequently, PE related to COVID-19
                    could be consider as triggered by a transient inflamma-     SOME PARAMETERS OF PLATELET FUNCTION IN
                    tory acute phase and treated for no longer than 3           COVID-19 PATIENTS
                    months. However, few data are available on manage-
                    ment of anticoagulation and risk of venous thromboem-       M. Scavone1, C. Ghali1, M. Calogiuri1, M. Sala2,




                                                                                                       ly
                    bolic (VTE) recurrences in these patients and guidelines    E. Bossi1, T. Mencarini3, S. Bozzi3, B. Clerici1,
                    are still undefined. Aim of the present study is to eval-   S. Birocchi1, N. Maugeri4, G. Marchetti2, M. Cattaneo1,




                                                                                               on
                    uate the long-term follow-up of a cohort of covid-19        G. Podda1
                    patients with PE. Methods: We conducted a retrospec-
                                                                                1Divisione di Medicina Generale II, ASST Santi Paolo e
                    tive multicenter study in four Italian hospitals between




                                                                                             e
                    March 1 st, 2020, and May 31st, 2021 in patients who        Carlo, DISS, Università degli Studi di Milano;
                                                                                2Dipartimento di Malattie Infettive, ASST Santi Paolo e
                    experienced a PE during hospitalization for a COVID-
                    19 pneumonia excluding patients who died during hos-
                    pitalization. Basal characteristics were collected and
                                                                                   us
                                                                                Carlo, Università degli Studi di Milano, Milan, Italy;
                                                                                3DEIB, Politecnico di Milano; 4Division of Immunology,

                    patients were grouped according to duration of antico-      Transplantation & Infectious Diseases, IRCCS San
                                                                                al
                    agulant treatment (≤3 months or >3 months). Primary         Raffaele Institute, Milan, Italy
                                                                        ci

                    outcome was incidence of VTE relapses, secondary out-
                    come was the composite of deaths, major hemorrhages         Background and Aims: Severe acute respiratory syn-
                                                                  er


                    and VTE relapses during follow-up. Results: 106             drome coronavirus 2 (SARS-CoV2) causes COVID-
                    patients with PE were discharged, of these 95 (89.6%)       19, an uncommon viral pneumonia commonly associ-
                                                             m



                    had a follow up longer than 3 months (seven patients        ated with a broad spectrum of presentations, including
                    were lost to follow up and four died within three           thrombotic and bleeding complications. The aim of this
                                                     om




                    months). The median follow-up was of 13 months (IQR         study was to characterize some parameters of the
                    1-19). Overall the 23.2% of subjects (22/95) was treated    platelet function in patients with COVID-19 without
                    up to three months or less whereas the 76.8% of them        (mild) and with pneumonia (severe). Methods: We
                    (73/95) received anticoagulation for more than three        studied 35 non-hospitalized patients with mild
                                                  -c




                    months. Basal characteristics are shown in Table 1.         COVID-19 (54% men; median age 67 (23-86) years),
                                                                                42 hospitalized patients with severe COVID-19 (62%
                                          on




                                                                                men, median age 64 (25-88) years), and 15 healthy
                                                                                controls (HC) (61% men, median age 42 (29- 65)
                                    N




                                                                                years). The following parameters were measured: 1)
                                                                                percentages of platelet/monocyte (PMAs) and
                                                                                platelet/granulocyte (PGAs) aggregates and of platelets
                                                                                expressing p-selectin on their membrane: these platelet
                                                                                activation markers were measured by flow cytometry
                                                                                in whole blood; 2) platelet aggregation (PA) induced
                                                                                by ADP 1 µM and Collagen 1 µg/mL in citrated
                                                                                platelet-rich plasma by light transmission aggregome-
                                                                                try (LTA); 3) platelet adenine nucleotides (ATP and
                                                                                ADP) content, by luminometer; 4) capacity of platelets
                                                                                to form thrombi on collagen (10 µg/mL)-coated
                                                                                microfluidic chambers perfused by whole blood at a
                                                                                shear rate of 300/s; D-dimer plasma levels, which were
                                                                                measured only in patients with severe COVID-19.
                    Table 1. Basal characteristics.
                                                                                Results: Median values of markers of in vivo platelet
                                                                                activation were increased in mild and severe COVID-
                    The 4.5% of patients in the short treatment group died      19 patients, compared to HC: PMAs, 16.5% vs 15.2%
                    in comparison to the 5.5% of patients in the long term      vs 8.2%, P<0.0001; PGAs, 7.1% vs 5.8% vs 5.2%,




                                                             [Hematology Reports 2021; 13:s3]                                             [page 89]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 90



         Poster




                     P=0.0355; p-selectin expression, 2.1% vs 3.2% vs               and Experimental Medicine, University of Foggia,
                     1.8%, P=0.0030. In patients with severe COVID-19,              Foggia, Italy; 5Multimedica Srl, Naples, Italy;
                     plasma D-dimer levels were increased and positively            4Department of Medicine and Health Sciences’VTiberio’,

                     correlated with PMAs (r=0.4433, P=0.0026) and PGAs             University of Molise, Campobasso, Italy
                     (r=0.5310,P=0.0002). The extent of PA induced by
                     exogenous agonists was heterogeneous in patients with          Background and Aims: The role of methylentetrahy-
                     mild and severe COVID-19; its median values were               drofolate reductase (MTHFR) C667T genotype, plasma
                     not significantly different from that of HC (ADP:              homocysteine (HC) and natural anticoagulants in portal
                     59.9% vs 36.5% vs 35.3%, P=0.7000; collagen: 75.7%             vein thrombosis (PVT) with liver cirrhosis (LC) derives
                     vs 72.3% vs 74.4%, P=0.3599). However, the lag time            from case-control studies comparing LC patients with
                     of collagen-induced platelet aggregation was signifi-          and without PVT. We compared age at PVT in a cohort
                     cantly longer in mild and severe COVID-19 than in              of LC patients genotyped for MTHFR C677T and iden-
                     HC. Platelets from patients with mild or severe                tified predictors of age at PVT. Methods: We recruited
                     COVID-19 showed a reduced capacity to form thrombi             128 LC patients with a PVT genotyped for MTHFR; we
                     on collagen-coated microfluidic chambers, compared             excluded 20 patients with other thrombotic risk factors
                     to HC (Table 1). Compared to HC, severe COVID-19               and investigated 108 LC participants (36 MTHFR CC,
                     patients had lower platelet ADP and ATP contents               53 MTHFR CT and 19 MTHFR TT); age, sex, age at
                     (ADP: 1.8 vs 3.5 nmol/108 platelets, P<0.0001; ATP:            PVT, plasma HC and natural anticoagulant concentra-
                     6.7 vs 7.9 nmol/108 platelets, P=0.0023) and higher            tions was available for all participants. Results: Age at




                                                                                                           ly
                     ATP/ADP ratios (4.1 vs 2.4, P<0.0001). There was a             PVT was lower and plasma HC was higher in MTHFR
                     statistically significant inverse correlation between          TT; antithrombin, protein C and S were lower in the




                                                                                                    on
                     plasma D-dimer levels and the platelet ADP content             MTHFR TT group (Table 1). MTHFR TT, male gender
                     (r=-0.3616, P=0.0279). Conclusions: In vivo platelet           and protein C predicted age at PVT (p-0.02, p=0.04 and
                     activation markers were increased in COVID-19                  p=0.08 respectively); MTHFR TT and Child-Pugh




                                                                                               e
                     patients, independently of the severity of the disease.        score positively predicted plasma protein C (p<0.0001
                     The median platelet ADP content was low in severe
                     COVID-19 patients, likely as a consequence of in vivo
                     secretion by activated platelets. COVID-19 patients
                                                                                        us
                                                                                    and p=0.0002 respectively) while plasma HC negative-
                                                                                    ly predicted protein C (p<0.0001). Age at PVT of the
                                                                                    MTHFR TT group was greater than that that of 19
                                                                                    al
                     displayed impaired collagen-induced PA by LTA and              patients with idiopathic PVT carrying the same geno-
                     formation of platelet thrombi on collagen-coated               type (56±13 vs 31.3±8.5, p<0.001) (cohort presented in
                                                                            ci

                     microfluidic chamber under controlled blood flow con-          separate abstract). Conclusions: MTHFR TT antici-
                     ditions.                                                       pates age at PVT in LC by an average of 9 years com-
                                                                      er


                                                                                    pared to MTHFR CC. HC adversely affects protein C
                     Table 1. Platelet parameters in patients with mild or severe   that plays a part in the premature PVT presentation in
                                                                m



                     COVID-19.                                                      the MTHFR TT setting.
                                                   om




                                                                                    Table 1.
                                                -c
                                        on
                                   N




                     PO015
                                                                                    PO016
                     PORTAL VEIN THROMBOSIS IN LIVER CIRRHOSIS
                     PATIENTS CARRYING THE HOMOZYGOUS MTHFR                         CANCER-ASSOCIATED VENOUS
                     C667T GENOTYPE DEVELOPS 9 YEARS EARLIER                        THROMBOEMBOLISM: THE ONCO-VTE START2 –
                     THAN IN WILD TYPE                                              REGISTRY

                     P.R.J. Ames1,2, G. D’Andrea3, A. Arcaro4, V. Marottoli5,       L. Barcella1, E. Antonucci2, L. Russo1, A. Chistolini3,
                     L. Iannaccone5, F. Gentile4, M. Maraglione3                    A. Serrao3, G. Elmi4, P. Sivera5, F. Schieppati1,
                                                                                    A. Tufano6, S. Pedrini7, G. Colombo8, W. Ageno8,9,
                     1Immune   Response & Vascular Disease Unit, NOVA               A. Falanga1,10, G. Palareti2
                     Medical School, Universidade NOVA de Lisboa, Lisboa,
                     Portugal; 2Dumfries & Galloway Royal Infirmary,                1UOC  Immunoematologia e Medicina Trasfusionale -
                     Dumfries, UK; 3Medical Genetics, Department of Clinical        Centro Emostasi e Trombosi, ASST Papa Giovanni




         [page 90]                                           [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 91



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    XXIII,      Bergamo;         2Fondazione        Arianna     ment of CAT is still limited. “Real life” data are needed
                    Anticoagulazione Bologna; 3Sezione Ematologia,              to monitor current clinical practice, to identify vari-
                    Dipartimento di Biotecnologie Cellulari ed Ematologia       ables associated to efficacy and safety of treatment and
                    Azienda Ospedaliero Universitaria Policlinico Umberto       to obtain suggestions for improving clinical practice
                    I, Roma; 4Dipartimento di Medicina Interna, Ospedale        and promote research.
                    Maggiore Bologna; 5SCDU Ematologia e Terapie                The Foundation has received an unrestricted research
                    cellulari AO Ordine Mauriziano di Torino; 6Centro di        grant from LEO Pharma S.p.A., specifically dedicated
                    Riferimento Regionale per le coagulopatie,                  to the realization of this study
                    Dipartimento di Medicina e Chirurgia, AOU Federico
                    II Napoli; 7UO Laboratorio Analisi, Dipartimenti di
                    Medicina di Laboratorio, Fondazione Poliambulanza           PO017
                    Istituto Ospedaliero, Brescia; 8DB Internistica e Centro
                    Trombosi ed Emostasi - AO di Circolo - ASST dei Sette
                                                                                INFERIOR VENA CAVA FILTER PLACEMENT IN
                    Laghi, Varese; 9Dipartimento Di Medicina e Chirurgia,
                    Università Degli Studi Dell’Insubria, Varese;               PATIENTS WITH ACUTE VENOUS
                    10Dipartimento di Medicina e Chirurgia, Università          THROMBOEMBOLISM: ANALYSIS OF CLINICAL
                    Milano-Bicocca, Monza, Italy                                INDICATIONS, ANTICOAGULATION MANAGEMENT
                                                                                AND IN-HOSPITAL MORTALITY
                    Background and Aim: Cancer-associated thrombosis




                                                                                                       ly
                    (CAT) is a major cause of morbidity and mortality in        D. Santagata1, G. Cammà2, A. Porfidia2, E. Porceddu2,
                    patients with malignancy. Anticoagulant treatment is        F. Piacentino3, F. Fontana3, M. Venturini3, W. Ageno1,




                                                                                                on
                    challenging because of the increased risk of recurrent      R. Pola2, M.P. Donadini1
                    venous thromboembolism (VTE) and bleeding.
                                                                                1Department   of Medicine and Surgery, Research Center
                    Information about the management of patients with




                                                                                          e
                    CAT in real life are still scanty. The purposes of the      on Thromboembolic Disorders and Antithrombotic
                    “VTE in the cancer patient (Onco-VTE) START2
                    (Survey on anTicoagulated pAtients RegisTer 2) regis-
                    ter are: 1) to prospectively collect data about CAT
                                                                                   us
                                                                                Therapies, University of Insubria, Varese, Italy;
                                                                                2Department of Internal Medicine, Fondazione

                                                                                Policlinico Universitario AGemelli IRCCS, Università
                                                                                Cattolica del Sacro Cuore, Rome, Italy; 3Department of
                                                                                al
                    patients, in particular the type of anticoagulant admin-
                    istered and the duration of treatment; 2) to collect        Diagnostic and Interventional Radiology, Ospedale di
                                                                        ci

                    information about the management of these patients in       Circolo e Fondazione Macchi, University of Insubria,
                    real life: the choice of anticoagulant drug and the dura-   Varese, Italy
                                                                  er


                    tion in relation to type of tumor, stage of disease,
                    comorbidities, associated therapies; 3) to evaluate the     Background: Inferior vena cava filters (IVCF) are cur-
                                                             m



                    incidence of thromboembolic and haemorrhagic com-           rently the most commonly used mechanical method to
                    plications of the anticoagulant treatment. Methods:         prevent embolization in patients with acute venous
                                                 om




                    The START2 is a multicenter, observational, independ-       thromboembolism (VTE) when an absolute contraindi-
                    ent ongoing register which prospectively collect clini-     cation to anticoagulation (AC) exists. However, the
                    cal information about patients on anticoagulant treat-      population requiring IVFC placement is highly het-
                                              -c




                    ment in “everyday clinical practice”. Data are collected    erogenous and still poorly characterized. Therefore,
                    by researchers of the participating centres by means of     there are no specific indications on how to restart AC
                                      on




                    an on-line electronic case report form. Onco-VTE is a       therapy after IVFC placement. Therefore, we aimed to
                    branch of the START2 register that includes additional      collect data on patients with acute VTE undergoing
                    information on the natural history of CAT and associ-       IVCF placement, in order to better characterize them
                                  N




                    ated oncological disease. Results: Presently, in Onco-      and provide information on their current in-hospital
                    VTE START2 register 122 CAT patient were enrolled;          management. Methods: A retrospective cohort study
                    45% males; median age (range) 66 (24;92) years; 48%         was conducted including consecutive patients objec-
                    with hematological cancer. In 52% of patients CAT           tively diagnosed with acute lower limb deep vein
                    occurred at the onset of cancer disease and in 33% of       thrombosis (DVT) and/or pulmonary embolism (PE)
                    patients CAT has been diagnosed incidentally. VTE           who underwent IVCF placement by interventional
                    events (79% deep venous thrombosis with or without          radiology service at Ospedale di Circolo, ASST Sette
                    pulmonary embolism; 21% isolated pulmonary                  Laghi (Varese) between 2015 and 2019, and at IRCCS
                    embolism) were treated in 52% of patients with direct       Fondazione Policlinico Universitario A. Gemelli
                    oral anticoagulant (DOAC); 4% of patients were treat-       (Rome) between 2010 and 2019. The objectives were
                    ed with Vitamin K Antagonist (VKA); 44% with low            to assess the characteristics of the population, includ-
                    molecular weight heparin (LMWH) or fondaparinux.            ing baseline characteristics, type of VTE events and
                    Concomitant use of antiplatelet drugs was recorded in       indication for IVFC placement, and to evaluate antico-
                    10% of patients. Conclusions: CAT patients are very         agulation management and in-hospital mortality.
                    heterogeneous. Several factors can influence the            Baseline characteristics and data on IVCF placement,
                    choice, duration, efficacy and safety of treatment of       AC management and in-hospital mortality were col-
                    CAT. Randomized clinical trials necessarily investigat-     lected from hospital databases. Results: We included
                    ed a sample of CAT patients and evidence for the treat-     317 patients undergoing IVCF placement: 143 women




                                                             [Hematology Reports 2021; 13:s3]                                               [page 91]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 92



         Poster




                     (45.1%) and 174 men (54.9%) with a mean age of 67,0        with MTS presenting with proximal DVT and treated
                     years (SD 15.7, range 14-99) (Table 1). Active cancer      with an endovascular procedure and long-term antico-
                     was present in 56.5% of patients and at least one          agulation. Materials and Methods: This study includ-
                     comorbidity, in addition to acute VTE, was present in      ed patients with a DVT secondary to MTS diagnosed
                     all patients, including 47.6% with hypertension, 16.4%     from March 2017 to May 2022 and treated with an
                     with diabetes mellitus and 12.6% with a history of         endovascular procedure. In all cases, MTS was diag-
                     VTE. The main reported indication for IVFC place-          nosed with CT venography or endovascular ultrasound
                     ment was recent bleeding (58.7%), followed by non-         scan. We collected clinical and radiological data: sex,
                     deferrable surgery (40.7%), high bleeding risk lesion      age, thrombophilic status, the pattern of DVT,
                     (21.5%), floating thrombus (3.8%) and anticoagulation      endovascular procedure time and type, adverse events,
                     failure (3.2%). Thirty-five patients (11,0%) died during   and status at last follow-up. Results: From March 2017
                     hospitalization after a mean of 31.7 days (SD 40) from     to May 2022, seven patients presented a DVT second-
                     IVFC placement. The main cause of death was sepsis         ary to an MTS. Patients characteristics are summarized
                     (34.3%) followed by bleeding (11.4%) and VTE               in Table 1. Six patients were female, and one was male.
                     (5.7%). At discharge, 218/282 patients (77.6%) were        The median age was 38 years old. All patients present-
                     receiving AC (20.6% parenteral at prophylactic dose,       ed a left leg proximal DVT with involvement of the
                     62.4% parenteral at intermediate/therapeutic dose,         proximal iliac axis; in one case, there was also an
                     7.8% DOACs and 4.2% VKAs). No significant associ-          extension to inferior vena cava (IVC). In one patient,
                     ation was found between filter indications and in-hos-     the diagnosis of MTS was made at the second relapse




                                                                                                         ly
                     pital mortality or AC initiation at discharge.             of DVT. Three patients (42%) presented a major
                     Conclusions: This study confirms that patients select-     thrombophilic status: one protein S deficiency, one




                                                                                                 on
                     ed for IVCF implantation are highly comorbid and           protein S deficiency + factor V Leiden, and one with
                     show significant short-term mortality. AC was started      combined factor V Leiden and factor II G20210A het-
                     in-hospital with parenteral drugs in the great majority    erozygosity. One patient showed heterozygosity for




                                                                                           e
                     of cases, most of them at therapeutic doses.               factor II G20210A mutation; the other three patients

                     Table 1.                                                       us
                                                                                had no abnormalities. All patients underwent an
                                                                                endovascular procedure. Two patients were treated in
                                                                                an acute phase (within 30 days of DVT diagnosis),
                                                                                while five patients were treated after median 17
                                                                                al
                                                                                months from the acute event. The procedure consisted
                                                                        ci

                                                                                of manual aspiration thrombectomy using a guide
                                                                                catheter, followed by balloon angioplasty and venous
                                                                   er


                                                                                stent placement. Only in one case, there was acute
                                                                                restenosis after two days, which required reoperation
                                                              m



                                                                                and placement of a new stent. Other two patients pre-
                                                                                sented anemia treated with blood transfusion in the
                                                  om




                                                                                first seven days after the procedure. No significant
                                                                                bleeding events have been recorded. All patients were
                     PO018                                                      treated with heparin for one month, followed by a long-
                                                                                term anticoagulant therapy with DOAC (apixaban in
                                               -c




                     ENDOVASCULAR TREATMENT OF DEEP VEIN                        five patients, rivaroxaban and edoxaban each in one
                                                                                patient). After a median follow-up of 18 months, no
                                        on




                     THROMBOSYS SECONDARY TO ILIAC VEIN
                     COMPRESSION (MAY THURNER SYNDROME):                        patients experienced a relapse of DVT or stent resteno-
                     A MONOCENTRIC CASE SERIES                                  sis with a significant improvement of the PTS.
                                   N




                                                                                Conclusions: It is important to suspect MTS in cases
                     G. Marchesini1, G. Carli1, I. Nichele1, L. Boi2,           of DVT in the left lower limb with involvement of the
                     A. Tosetto1                                                iliac axis and resistant to conventional anticoagulant
                                                                                therapy. In these cases, the endovascular approach rep-
                     1Hemophilia  and Thrombosis Center, Hematology             resents a viable therapeutic option with high success
                     Department, SBortolo Hospital, Vicenza; 2Complex           rates, especially in mitigating the severity of the PTS.
                     Operational Unit of Radiology, SBassiano Hospital,
                                                                                Table 1. Patients’ characteristics (DVT=deep vein thrombosys,
                     Bassano del Grappa, Italy
                                                                                PTS= post thrombotic syndrome).
                     Background: May-Thurner syndrome (MTS) is a rare
                     clinical entity featuring venous obstruction of the left
                     lower leg that can cause deep vein thrombosis (DVT)
                     resistant to anticoagulant therapy with a high risk of
                     severe post-thrombotic syndrome (PTS). There are no
                     standard guidelines to treat DVT secondary to MTS or
                     its long-term sequelae, but endovascular treatment
                     appears to be a promising option in selected cases.
                     Aim: We aimed to describe a case series of patients




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 93



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    PO019                                                        shifted to sulodexide (20.1%), and 4.9% pt/y (including
                                                                                 one MB) in those shifted to antiplatelets (7.3%)
                                                                                 (p=0.23). Conclusions: The provoked/unprovoked
                    OUTCOMES DURING FOLLOW-UP IN VENOUS                          nature of the index event was not the prevalent criterion
                    THROMBOEMBOLISM PATIENTS INCLUDED IN                         for duration of treatment. Anticoagulation was contin-
                    THE INTERNATIONAL, PROSPECTIVE,                              ued in more than half of cases, whereas treatment was
                    OBSERVATIONAL WHITE STUDY                                    shifted to different antithrombotic agents in 27.4% of
                                                                                 patients and the recommendation to stop any specific
                    M. Cini1, T. Urbanek2, Y.J. Li3, J. Madaric4,                treatment was given to about 20% of all patients.
                    K. Bouslama5, G.Y. Sokurenko6, G.M. Andreozzi7,              Almost all patients in whom anticoagulation was
                    J. Matuška8, A. Mansilha9, V. Barinov10, G. Palareti1,       extended were treated with DOAC. Only in Tunisia
                    for the WHITE study group (Fondazione Arianna                DOACs were not available when the study was real-
                    Anticoagulazione)                                            ized. During follow up, one MB occurred among
                    1Fondazione Arianna Anticoagulazione,
                                                                                 patients treated with antiplatelet drugs, while the rate of
                                                            Bologna, Italy;      VTE recurrences was in general low in all patients. The
                    2Katowice,   Poland; 3Peking Union Medical College,          rate of primary events was the highest in patients treat-
                    Beijing, China; 4Comenius University, Bratislava,            ed with antiplatelets.
                    Slovakia; 5Faculty of Medicine of Tunis, Tunisia;
                    6StPetersburg, Russia; 7University of Padua, Italy;




                                                                                                         ly
                    8Clinical Trial Centre, Hodonin, Czechia; 9University of
                                                                                 PO020
                    Porto, Portugal; 10Clinical hospital N1 “Volynskaya”,




                                                                                                  on
                    Moscow, Russia
                                                                                 CATHETER-RELATED RIGHT ATRIAL THROMBOSIS
                    Background and Aims: How to prevent recurrencies             IN CANCER PATIENTS: A RETROSPECTIVE




                                                                                            e
                    after a first venous thromboembolism (VTE) may be            ANALYSIS
                    influenced by many factors. The WHITE study was an
                    international, prospective, observational, investigators-
                    initiated, no profit study (ClinicalTrials.gov Identifier:
                                                                                     us
                                                                                 E. Porceddu2, E. del Sordo1, A. D’Errico1, A. Porfidia2,
                                                                                 P. Tondi2, A. Gasbarrini1, R. Pola2
                                                                                 al
                    NCT04646993) promoted by Fondazione Arianna
                                                                                 1Department     of Internal Medicine, Fondazione
                    Anticoagulazione (Bologna, Italy), that aimed at ana-
                                                                         ci

                    lyzing a) the treatment decisions taken on this issue in     Policlinico Universitario AGemelli IRCCS, Università
                    clinical practice in various countries, different for        Cattolica del Sacro Cuore, Rome, Italy; 2Department of
                                                                   er


                    socio-economic conditions and healthcare systems, and        Geriatrics, Neurosciences and Orthopedics, Fondazione
                    b) the clinical outcomes recorded during one year fol-       Policlinico Universitario AGemelli IRCCS, Università
                                                             m



                    low-up (FU) after the treatment decisions were taken         Cattolica del Sacro Cuore, Rome, Italy
                    (which is the aim of this presentation). Methods: 1004
                                                 om




                    patients (510 females), with a prior first VTE event,        Background and Aims: Literature data on catheter-
                    were included in the study in 62 clinical centers in 7       related right atrial thrombosis (CRAT) are scarce and
                    countries from April 2018 to December 2020 [China (n.        based only on case reports or case series1-2. Data mostly
                                                                                 concern dialysis patients while reports on oncological
                                              -c




                    258), Czechia (n. 67), Poland (n. 128), Portugal (n. 33),
                    Russia (n. 415), Slovakia (n. 23), and Tunisia (n. 80)],     patients are poor2. In this study we investigated charac-
                                       on




                    and followed-up for one year after receiving a decision      teristics of cancer patients with CRAT to provide a better
                    about treatment beyond the initial and maintenance           characterisation and to improve the knowledge of this
                    phases. Results: The index event was unprovoked in           disease. Methods: We retrospectively analysed data of
                                  N




                    57.9% of patients and provoked in the remaining.             21 consecutive cancer outpatients with CRAT that
                    During FU, 53.4% of patients continued anticoagula-          referred to our Internal Medicine and Thromboembolic
                    tion, 27.4% continued with antiplatelets, 19.2% stopped      Pathology Unit over a period of 3 years. Of these
                    all treatments. When anticoagulation was continued, a        patients we collected epidemiological and clinical data
                    direct oral anticoagulant (DOAC) was used in 88.6%,          and evaluated the efficacy (resolution of the thrombotic
                    vitamin K antagonist (VKA) in 8.4% and low molecu-           process and recurrences) and safety (bleedings) of the
                    lar weight heparin (LMWH) in 3.0% of cases                   different anticoagulant treatments used. Results: In our
                    (p<0.001). Anticoagulant treatment continued in 53.0%        study, the median age was 57 years. There were 4 (19%)
                    of patients with unprovoked and in 53.9% with pro-           males and 17 (81%) females. All patients had active neo-
                    voked events (p=0.78). Anticoagulation was continued         plasia: 8 (38%) ovarian, 5 (24%) uterine, 5 (24%) col-
                    in: 59.0% of patients with proximal (±distal) deep vein      orectal, and 3 (14%) breast cancer. Of them, 15 (71%)
                    thrombosis (DVT), 39.0% of isolated distal DVT,              were metastatic. In 5 (24%) cases, thrombosis was pres-
                    65.0% of cases with pulmonary embolism (PE) with or          ent at other sites, in addiction to the right atrium, i.e.,
                    without DVT (p<0.001). Primary outcomes (recurrent           pulmonary embolism (PE) and deep veins of the upper
                    VTE, and major bleed-MB) during FU occurred in:              limbs. In 2 cases, the diagnosis of CRAT was made by
                    2.9% pt/y in patients who extended the anticoagulation       echocardiography, while in the other 19 cases by CT-
                    with DOAC (47.3% of the total), 2.2% pt/y when all           angiography, with further confirmation by echocardiog-
                    treatments were stopped (19.2%), 3.5% pt/y in patients       raphy in 15 patients. The median follow-up was 10




                                                              [Hematology Reports 2021; 13:s3]                                                 [page 93]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 94



         Poster




                     months. Nineteen (90%) patients received anticoagulant        Background and Aims: The optimal management
                     treatment for 3-6 months: 17 with low molecular weight        strategy for prevention of recurrent deep vein throm-
                     heparin ad 2 with direct oral anticoagulants (DOACs).         bosis (DVT) is still uncertain. Clinical and laboratory
                     After completion of the treatment period, 9 (43%)             assays and imaging tests such us residual vein throm-
                     patients extended treatment with low dose anticoagula-        bosis (RVT) have been proposed as risk factors for
                     tion. In 12 (57%) patients there was complete resolution      recurrence. Though, which patients have a higher risk
                     of CRAT, in 5 (24%) a partial resolution, and in 3 (14%)      of RVT is still unclear. Some categories have been
                     the thrombus remained stable. Data are not available in       identified, such as the male gender, obese and cancer
                     1 (5%) patient. No major bleedings occurred, nor clini-       patients or the extensive thromboses. Furthermore,
                     cally relevant non major bleedings. There were only 4         most of data are from vitamin K antagonists (VKA)
                     minor bleedings: 2 epistaxis and 2 genital bleedings.         treatment and experience on direct oral anticoagulants
                     One pulmonary embolism occurred during anticoagu-             (DOACs) is scanty. Therefore, we designed a prospec-
                     lant treatment. Conclusions: With the increase of             tive study to evaluate which clinical features are more
                     patients with central venous catheters, CRAT is becom-        predictive of pathological RVT during DOACs treat-
                     ing less uncommon than previously thought. Early treat-       ment. Methods: The local Ethics Committee approved
                     ment may lead to complete thrombosis resolution in            our study, that we called READ (“residual veins
                     more than half of patients. Anticoagulants are safe with      thrombosis under DOACs”), in November 2019. All
                     low risk of bleeding. The risk of recurrences is similar to   the eligible patients signed an informed consent based
                     the risk displayed by thrombosis in usual sites in cancer     on the Declaration of Helsinki. As from literature,




                                                                                                          ly
                     patients3-4.                                                  RVT was defined as persistent thrombotic material in
                                                                                   a diameter of at least 4 mm, identified by compression




                                                                                                   on
                     References:                                                   ultrasonography (CUS) at common femoral or
                      1. Aristeidis Stavroulopoulos, Vasiliki Aresti et al.        popliteal veins. Patients have been evaluated at 6
                         Right atrial thrombi complicating haemodialysis           weeks (T1), 3 months (T2) and 6 months (T3) through




                                                                                             e
                         catheters. A meta-analysis of reported cases and a        clinical and demographic variables. The study is still
                         proposal of a management algorithm Nephrol Dial
                         Transplant (2012) 27: 2936–2944
                      2. Minh-Ha Tran1,2, Tanya Wilcox et al. Catheter-
                                                                                      us
                                                                                   ongoing and we have so far enrolled 80 patients. We
                                                                                   are here presenting the interim results on a total of 50
                                                                                   patients who completed a 6-month follow up in which
                                                                                   al
                         related right atrial thrombosis. The Journal of           they received a direct oral anticoagulant (DOAC).
                         Vascular Access 2020, Vol. 21(3) 300–307                  Results: In our cohort of patients, composed of 66%
                                                                           ci

                      3. Prandoni P, Lensing AW, Piccioli A, et al. Recurrent      and 34% of respectively male and female patients,
                         venous thromboembolism and bleeding complica-             54% was treated with Apixaban, 40% with
                                                                     er


                         tions during anticoagulant treatment in patients with     Rivaroxaban, 4% with Edoxaban and 2% with
                         cancer     and     venous     thrombosis.     Blood       Dabigatran. RVT rate was 77.1% at six weeks, 58% at
                                                               m



                         2002;100:3484-8                                           three months and 43.5% at six months. Male gender
                      4. T.F. Wang, A. Li, et al. Managing thrombosis in           was independently associated with RVT at six weeks
                                                   om




                         cancer patients. Res Pract Thromb Haemost, 2 (3)          (OR 4.75, CI 1.13 – 19.68, p=0.036), three months
                         (2018), pp. 429-438                                       (OR 6.4, CI 1.75 – 23.35, p=0.006) and six months
                                                                                   (OR 7.71, CI 1.48 – 40.24, p=0.011). In the univariate
                                                -c




                                                                                   model, DVT of at least two segments was associated
                     PO021                                                         at higher RVT rate at six weeks (OR 5.29, CI 1.00 –
                                        on




                                                                                   27.93, p=0.045) and three months (OR 3.54, CI 1.07 –
                                                                                   11.78, p=0.047) and to greater RVT extension at six
                     RESIDUAL VEIN THROMBOSIS AND PREDICTIVE                       months (5.4 mm±2.7 mm vs 3.6 mm±1.7 mm,
                                   N




                     FACTORS OF RECANALIZATION UNDER DIRECT                        p=0.016). Diagnostic delay of at least one month was
                     ORAL ANTICOAGULANTS. A SINGLE CENTER                          associated to higher rate of RVT at six months (OR
                     PROSPECTIVE STUDY                                             6.46, CI 1.17 – 35.74, p=0.029) and to greater RVT
                                                                                   extension at three months (7.3±3.6 vs 4.8±2.8,
                     M. Bardetta1, A. Pizzuto2, M. Simoncini3,                     p=0.02). Obesity was associated to lower rate of RVT
                     P. Martinelli1, C. Sella1, C. Dainese1, F. Valeri1,           at three months (OR 0.12, CI 0.22 – 0.65, p=0.011)
                     B. Bruno4, A. Borchiellini1                                   and at six months (OR 0.1, CI 0.01 – 0.87, p=0.03).
                                                                                   Main results are shown in Figure 1. Conclusions:
                     1University  Division of Hematology, Regional Center          Male gender is confirmed as independent predictor for
                     for Hemorrhagic and Thrombotic Diseases, City of              persistence of RVT in patients treated with DOACs. In
                     Health and Science University Hospital of Molinette,          our experience, extension of DVT is associated to a
                     Turin, Italy; 2University Division of Internal Medicine       reduced recanalization as previously described during
                     2“GMontrucchio”, City of Health and Science                   VKA treatment. Diagnostic delay as well seem to be
                     University Hospital of Molinette, Turin, Italy;               associated to a reduced recanalization. On the contrary
                     3Emergency Medicine Division, City of Health and              obesity does not seem to influence negatively the
                     Science University Hospital of Molinette, Turin, Italy;       recanalization. Unlike previous reports, prevalence of
                     4University Division of Hematology, City of Health and        RVT in our cohort is similar to published experience
                     Science University Hospital of Molinette, Turin, Italy        in VKA patients.




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                                                                      XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                              patients at time T2 of follow up in DOAC therapy.
                                                                              Reliability between internal (CUS) and external (DU)
                                                                              observations has been computed according to Cohen’s
                                                                              kappa coefficient. In our cohort, RVT rate at T1 was
                                                                              77%, at T2 58% and at T3 43%. Comparing results
                                                                              obtained at the same time in the two different settings,
                                                                              CUS versus DU, for an amount of 84 total observa-
                                                                              tions, 21 out of 84 (25%) have discordant results. The
                                                                              degree of agreement between CUS and DU was just
                                                                              over “moderate” (κ=0.42, p=<0.0001). Only 27.9%
                                                                              (19 out of 68) of all positive DU were confirmed at
                                                                              CUS. On the other hand, the deviation was only 12.5%
                                                                              (2 out of 16) in case of negative DU (Figure 1). Low
                    Figure 1.
                                                                              level of agreement shown between DU performed by
                                                                              independent specialists without defined specific crite-
                                                                              ria and our CUS evaluations, highlights the impor-
                    PO022
                                                                              tance of a shared and reproducible elements to define
                                                                              RVT for clinical use. Indeed, the main difference con-
                    CAN RESIDUAL VEIN THROMBOSIS GUIDE                        sists of RVT smaller than 4 mm, so insufficient to be




                                                                                                     ly
                    CLINICAL PRACTICE AND THERAPY IN PATIENTS                 considered as CUS positive but well documented in
                                                                              DU. Highest agreement in negative reports may sug-




                                                                                              on
                    WITH PREVIOUS THROMBOTIC EVENT?
                    COMPARISON BETWEEN CUS AND DOPPLER US                     gest a well negative predictive value of CUS. The role
                    DATA FROM “READ” MONOCENTRIC                              of CUS to evaluate RVT and to guide clinical practice
                    PROSPECTIVE STUDY                                         and therapy is still debated but first analyses show




                                                                                          e
                                                                              possible important future developments in this field,
                    A. Pizzuto, M. Simoncini, P. Martinelli, M. Bardetta,
                    C. Sella, C. Dainese, F. Valeri, B. Bruno,
                    A. Borchiellini
                                                                                  us
                                                                              also thanks to the large spreading of CUS among cli-
                                                                              nicians.
                                                                              al
                    Università degli Studi di Torino, Italy
                                                                       ci

                    The advent of new therapeutic strategies, i.e. the
                                                                 er


                    Direct Oral Anticoagulants (DOACs), to treat Deep
                    Vein Thromboses (DVT), rises questions concerning
                                                              m



                    both timing and dose. In patients with a previous DVT,
                    Residual Vein Thrombosis (RVT) is taking more and
                                                 om




                    more ground as a possible risk factor of recurrence,
                    and on the other hand as a prognostic tool to guide
                    clinical practice. In this context, Compression           Figure 1.
                                              -c




                    Ultrasonography (CUS) could be applied to assess
                    RVT thanks to its simplicity, accessibility, even in
                                       on




                    bedside setting, and increasingly widespread use          PO023
                    among non-radiology clinicians. We designed a
                    prospective study, named READ (“RVT in DOAC
                                  N




                                                                              UPDATED SURVEY ON TREATMENT AND
                    therapy”), to evaluate the predictive factors of patho-
                                                                              SECONDARY PROPHYLAXIS OF VENOUS
                    logical RVT and, secondly, we are also verifying reli-
                    ability between our CUS determination and external        THROMBOEMBOLISM WITH ANTI XA IN PATIENTS
                    center Doppler Ultrasonography (DU) reports. We are       WITH SEVERE HEREDITARY THROMBOPHILIA-
                    here presenting the latter results. The Ethics            THE EXPERIENCE OF THREE SOUTHERN ITALY
                    Committee approved our study in November 2019. All        CENTRES EXPERIENCE
                    patients signed an informed consent based on the
                    Declaration of Helsinki. Patients have been evaluated     P. Madonna1, A. Guida1, A. De Sena1, P. Tirelli,1
                    at T1 (6 weeks), T2 (3 months) and T3 (6 months)          A. Vitale2, G. Papa2, S. Mangiacapra3, M. Amitrano3,
                    through clinical and demographic variables and CUS.       G. Landi2 C. Bologna1, M.G. Coppola1, N. Vitelli2,
                    This technique consists in the evaluation, under com-     A. Tufano2
                    pression, of two major thrombogenic sites of DVT of
                                                                              1UOC  Medicina Interna Ospedale del Mare ASL Napoli
                    lower limbs: the common femoral vein (from great
                    saphenous to subsartorial vein) and popliteal vein        1 Centro; 2Dipartimento di Medicina Clinica e
                    (from very proximal tract to tibial vein). We defined     Chirurgia AOU Federico II; 3UOC Medicina Interna
                    CUS test positive when RVT measured at least 4 mm         AORN GMoscati, Avellino, Italy
                    in transverse diameter, as widely accepted. The study
                    is still ongoing and here are the interim results of 50   Background and Aims: Severe Inherited throm-




                                                              [Hematology Reports 2021; 13:s3]                                           [page 95]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 96



         Poster




                     bophilia (SIT): antithrombin (AT), protein-C (PC),       PO024
                     protein-S (PS) deficiency, homozygousity or com-
                     pound heterozygosity Factor-II (FII)-G20210A and
                                                                              IDIOPATHIC PORTAL VEIN THROMBOSIS IN
                     FV-Leiden (FVL) polymorphism although rare,
                     increases the risk of venous thromboembolism             HOMOZYGOUS MTHFR C667T CARRIERS
                     (VTE), annual incidence >1%. A paucity of data is        DEVELOPS 20 YEARS EARLIER THAN IN WILD
                     available in VTE treatment with direct oral anticoag-    TYPE
                     ulants (DOACs) in these setting, aim of our survey
                     was to observe the efficacy and safety of DOACs in       P.R.J. Ames1,2, G. D’Andrea3, A. Arcaro4, V. Marottoli5,
                     SIT. Methods: 36-subjects (19M;17F, mean                 L. Iannaccone5, M. Maraglione3, F. Gentile4
                     age:40,89) with VTE: confirmed diagnosis of acute        1Immune Response & Vascular Disease Unit, NOVA
                     deep vein thrombosis (DVT) and/or pulmonary
                     embolism (PE), referred between March 2014 and           Medical School, Universidade NOVA de Lisboa, Lisboa,
                     March 2022 to three Campania thrombosis-centres,         Portugal; 2Dumfries & Galloway Royal Infirmary,
                                                                              Dumfries, UK; 3Medical Genetics, Department of
                     were enrolled. All patients were prescribed DOACs
                                                                              Clinical and Experimental Medicine, University of
                     for at least 3 months. All subjects were screened for
                                                                              Foggia, Foggia, Italy; 4Department of Medicine and
                     AT, PC, PS deficiencies, FVL, FIIG20210A polymor-
                                                                              Health Sciences ‘VTiberio’, University of Molise,
                     phisms. Hyperhomocysteinaemia and antiphospho-
                                                                              Campobasso, Italy; 5Multimedica Srl, Naples, Italy
                     lipids were excluded. VTE was treated with anti-Xa




                                                                                                       ly
                     anticoagulants. 3-months clinical follow-up was          Background and Aims: Studies on idiopathic portal




                                                                                               on
                     scheduled. Results: We identified 4 AT-deficiencies,     vein thrombosis (PVT) and inherited thrombophilia are
                     2 PC-deficiencies, 7 PS-deficiencies, 13 combined        mostly case-control; in our cohort we 1) compared age at
                     heterozigosity for FVLeiden and FIIG20210A, 4 FVL        PVT and plasma homocysteine (HC) in PVT patients
                     homozigosity, 3 FII-G20210A homozigosity and 2




                                                                                         e
                                                                              devoid of circumstantial factors for PVT, genotyped for
                     combined FVL-heterozigosity and FIIG20210A
                     homozygosity. 22/36 received Rivaroxaban, 5/36
                     Edoxaban and 9/36 Apixaban; 9/36 switched from
                     VKA to anti-Xa. 28/36 were prescribed lifelong full-
                                                                                  us
                                                                              methylentetrahydrofolate reductase (MTHFR) C667T; 2)
                                                                              identified predictors of age at PVT. Methods: 298 PVT
                                                                              patients were genotyped for MTHFR; we excluded 212
                                                                              patients with different risk factors for thrombosis, includ-
                                                                              al
                     dose therapy, two patients withdrawn therapy after 12    ing factor V Leiden, protein C and S deficiency. Of the
                     and 24 months respectively. During the 8-years fol-      remaining 86 participants, 24 were MTHFR CC, 45
                                                                      ci

                     low-up, neither thrombotic recurrences nor major         MTHFR TC and 17 MTHFR TT; age, sex, age at PVT,
                                                                 er


                     hemorrhagic events were observed. No differences in      smoking status, plasma HC and natural anticoagulant
                     sex, type of SIT, or drug. (Table 1) Conclusions: Our    concentrations were recorded. Results: Age at PVT was
                     survey suggests efficacy and safety of anti-Xa antico-
                                                           m



                                                                              lower and plasma HC was higher in MTHFR TT than in
                     agulants for treatment and secondary prophylaxis of      other genotypes; natural anticoagulant concentrations
                                                om




                     VTE in patients with SIT. Furthermore extensive mul-     were similar (Table 1).
                     ticentric studies are needed.
                                                                              Table 1. Demographics, clinical and laboratory variables by
                     Table 1.                                                 MTHFR genotypes.
                                             -c
                                      on
                                  N




                                                                              MTHFR TT and protein C predicted age at PVT
                                                                              (p<0.0001 & p=0.04 respectively); MTHFR TT predict-
                                                                              ed plasma HC (p=0.05). Smoking and sex predicted
                                                                              cavernoma (p=0.002 and p=0.02 respectively). Plasma
                                                                              HC inversely related to protein C in the MTHFR TT
                                                                              group (r=-58, p=0.01). Age at PVT of the MTHFR TT
                                                                              group was lower than that that of 19 PVT with liver cir-
                                                                              rhosis carrying the same genotype (31.3±8.5 vs 56±13,
                                                                              p<0.001) (cohort presented in separate abstract).
                                                                              Conclusions: MTHFR TT anticipates age at PVT by an




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    average of 20 years compared to other MTHFR geno-            AngioCT performed on day 7 resulted in a really good
                    types; protein C predicts age at PVT and is adversely        initial response with partial recanalization of left sig-
                    affected by plasma HC; smoking is associated with cav-       moid and transverse sinus, complete recanalization of
                    ernoma.                                                      right transverse sinus; subdural effusion stable. The
                                                                                 patient was discharged after 26 days with a prophylactic
                                                                                 dosage of LMWH, still ongoing. Currently the patient is
                    PO025                                                        receiving Eltrombopag 50 mg OD. A further MRI con-
                                                                                 trol will be performed within 2 months, after 3 months
                                                                                 of anticoagulant. Conclusions: We report a rare episode
                    CEREBRAL VENOUS SINUS THROMBOSIS IN A
                                                                                 of CSVT in a patient with XLT during therapy with
                    PATIENT WITH WISKOTT-ALDRICH SYNDROME
                                                                                 Eltrombopag, possibly related to a concomitant febrile
                    DURING THERAPY WITH ELTROMBOPAG                              episode of unknown origin and a sudden large increase
                                                                                 of the platelet. Even if Eltrombopag could have had a
                    T. Martini1, P. Pedrazzi1, L.D. Notarangelo2, E. Bertoni3,   causative role as already reported in literature, it was
                    R. Biguzzi1, C. Biasoli1                                     reintroduced to provide a safe and adequate platelet for
                    1Centro
                                                                                 LMWH treatment. A close radiological control showed
                              MEC Romagna, Medicina Trasfusionale                a good response, allowing us to reduce LMWH to a pro-
                    Ospedale Bufalini Cesena; 2Direzione medica, Presidio        phylactic dosage that should be safe and effective until
                    Ospedale dei Bambini, ASST-Spedali Civili, Brescia;          the complete resolution of the thrombosis.




                                                                                                          ly
                    3U.OOncoematologia Pediatrica, ASST-Spedali Civili,

                    Brescia, Italy




                                                                                                  on
                    Background and Aims: Wiskott-Aldrich Syndrome
                    (WAS) is a rare X-linked immunodeficiency character-




                                                                                             e
                    ized by microthrombocytopenia, eczema, recurrent
                    infections and autoimmunity. In its classical form
                    HSCT or gene Therapy are the treatments of choice. Its
                    milder form (X-linked Thrombocytopenia, XLT) could
                                                                                     us
                    be managed conservatively. Eltrombopag, a TPO-
                                                                                 al
                    receptor agonist, has been used in patients with Immune
                                                                         ci

                    Thrombocytopenia as well as in WAS/XLT but rare
                    cases of atypical venous thrombosis have been report-
                                                                   er


                    ed. Here we describe a case of a young patient with
                    XLT, in therapy with Eltrombopag, who developed a            Figure 1.
                                                             m



                    Cerebral Venous Sinus Thrombosis (CSVT) concomi-
                    tantly with a febrile episode. Methods: We evaluated
                                                 om




                    the medical record of the patient, collecting data about     PO026
                    clinical history, laboratory results and radiological
                    exams. Results: A 19-years old XLT patient started
                    treatment with Eltrombopag in 2018 (base platelet            IS HYPER-HOMOCYSTEINE AN INDEPENDENT
                                              -c




                    value around 15.000/mmc) with an effective dose of 50        RISK FACTOR FOR THROMBOSIS?
                                                                                 RETROSPECTIVE ANALYSIS OF A SINGLE-
                                       on




                    mg OD (platelet count at steady state around
                    50.000/mmc) in absence of bleeding. One month ago he         CENTRE PATIENTS’ POPULATION
                    went to Emergency Room for the onset of headache
                                  N




                    since 3 days, not sensitive to analgesics, and fever,        G.M. Nicolò1, G. Sottilotta1, F. Luise2, A. Piromalli2
                    without any other symptom. Complete blood count
                    showed 184.000 platelet/mmc. A Covid swab was neg-           1Hemophilia    Centre - Thrombosis and Hemostasis
                    ative. Cerebral AngioCT documented a complete                ServiceGreat Metropolitan Hospital of Reggio Calabria,
                    thrombosis of sigmoid sinus and jugular bulb, involving      Italy; 2Analysis LaboratoryGreat Metropolitan Hospital
                    the proximal third of the internal jugular vein, accom-      of Reggio Calabria, Italy
                    panied by a thin subdural hemorrhagic effusion.
                    Suddenly after the exam he had an episode of general-        Background and Aims: Homocysteine (HCY) is
                    ized seizure. Anticoagulation with LMWH (Low                 formed from the dietary amino acid methionine and plays
                    Molecular Weight Heparin was promptly started and            a pivotal role in folate metabolism and methyl group
                    Eltrombopag suspended for 5 days (platelet count             transfer. Hyper homocysteine (hHCY) is associated with
                    124.000/mmc). To maintain an adequate platelet count         arterial and venous thrombosis via several mechanisms
                    for safe anticoagulation it was gradually reintroduced       such as enhanced platelet reactivity, increased tissue fac-
                    (25 mg OD on day 5, incremented to 50 mg on day 7)           tor expression and thrombin generation, and endothelial
                    with a platelet count ranging between 35.000 and             dysfunction. The aim of our study was to evaluate if
                    45.000/mmc (Figure 1); this strategy allowed us to use       hHCY may be considered as an independent thrombosis
                    a therapeutic dosage of LMWH for the first 3 days, fol-      risk factor and its incidence in different sites of thrombo-
                    lowed by a subtherapeutic dosage (50 U/Kg BID). An           sis. Methods: Clinical data of 319 patients were retro-




                                                              [Hematology Reports 2021; 13:s3]                                                  [page 97]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 98



         Poster




                     spectively analysed, 184 with hHCY and 135 with nor-            1 Centro; 2Dipartimento di Medicina Clinica e
                     mal HCY (nHCY), followed by our centre during the last          Chirurgia AOU Federico II Napoli, Italy
                     6 years. We excluded any other conditions of throm-
                     bophilia: the Factor V Leiden and Prothrombin G20210            Background: Direct oral anticoagulants (DOAC) are
                     polymorphisms, Protein C and S deficiency, presence of          currently recommended as initial treatment of Venous
                     lupus anticoagulant and increased anticardiolipin or anti-      Thromboembolism (VTE). Although the evidence on
                     beta2 glycoprotein-I antibody levels. Results: 29/184           the use of these drugs in patients with unusual-site
                     with hHCY were found with thrombosis. Of these: 18/29           venous thromboembolism (VTE) is limited to a few,
                     (62.1%) had venous thrombosis (DVT), 11/29 (37.9%)              small studies, these drugs are currently used in clinical
                     had arterial thrombosis. We considered three hHCY               practice. Aim of our survey was to evaluate the efficacy
                     severity levels: mild 15-29 μmol/L, moderate 30-100             and safety of anti Xa-direct oral anticoagulants (aXa-
                     μmol/L and severe>100 μmol/L. 22/135 (16.3%) nHCY               DOAC s) in treatment and secondary prevention of
                     patients were diagnosed with thrombosis. Of these:              VTE in 9 non neoplastic patients. Methods: We
                     17/135 (12.6%) had DVT and 5/135 arterial thrombosis            enrolled 9 patients (2F, 7 Mmean age 53,44) referred to
                     (3.7%). Even in this case, we observed more DVT                 our thrombosis centre since march 2019 to march 2022.
                     (17/22,77.3%) than arterial thrombosis (5/22,22.7%).            All patients were screened for inherited and acquired
                     The comparison about incidence of thrombosis between            thrombophilia, Jak-2 polymorphism, autoimmunity and
                     hHCY and nHCY groups evidenced similar percentages              neoplasm. Diagnosis of VTE was obtained with con-
                     (15.8% and 16.3% respectively). Results are resumed in          trast-CT-Scan. 5 patients experienced splanchnic vein




                                                                                                            ly
                     Table 1. We found different thrombosis percentages com-         thrombosis, in 3 cases secondary to abdominal infection
                     paring the subjects with normal and mild hHCY to those




                                                                                                     on
                                                                                     (diverticulitis, cholecystitis) 3 patients had vena cava
                     with moderate hHCY: people with moderate hHCY had               thrombosis, 2 cases were provocked VTE (1
                     a much higher percentage of arterial thrombosis (61.5%)         spondilodiscitis, sepsis and inferior vena atresia) 1
                     than those with nHCY (22.7%) and patients with mild             patients presented FII-G20210A heterozigousity, 1




                                                                                               e
                     hHCY (15.8%), even if these data were not sufficient to         patient Factor V Leiden heterozigousity no other throm-
                     hypothesize a correlation between levels of HCY and
                     sites of thrombosis. Conclusions: Our analysis did not
                     demonstrate that hHCY can be considered an independ-
                                                                                        us
                                                                                     bophilic abnormalities were detected. All patients per-
                                                                                     formed a Total body CT-Scan and endoscopy was done
                                                                                     in 5/9 patients no solid tumors emerged. 3/9 were pre-
                                                                                     al
                     ent risk factor for thrombosis. Previous reports have           scribed apixaban the others received edoxaban.
                     shown that hHcy-promoting thrombotic events require a           Conclusions: Neither thrombotic events nor haemor-
                                                                             ci

                     first hit, such as dyslipidemia or hypertension. In our         rhagic complication were observed in this 3 years fol-
                     opinion, as reported by other authors, hHCY could be            lowup. All patients were prescribed longterm anticoag-
                                                                      er


                     only considered as a prothrombotic cofactor. Further            ulant therapy after informed consent. In a our small case
                     prospective studies with dedicated designs are needed to        series we observed effectiveness and safety of anti Xa-
                                                                m



                     clarify if hHCY may play an independent prothrombotic           DOACs in patients with atypical site vein thrombosis
                     role in the etiopathogenesis of thrombosis.
                                                    om




                     Table 1.
                                                                                     PO028
                                                 -c




                                                                                     A CASE OF TRICUSPID VALVE LIBMAN-SACKS
                                                                                     ENDOCARDITIS IN A PREGNANT WOMAN WITH
                                         on




                                                                                     PRIMARY ANTIPHOSPHOLIPID SYNDROME
                                    N




                                                                                     S. Migliorini1, C. Santoro2, A. Scatteia3,
                                                                                     S. Dellegrottaglie4, V. Cuomo5, E. Pilato6,
                                                                                     G. Comentale6, M. D’armiento7, M. Guida1, L. Sarno1,
                                                                                     A. Tufano5
                                                                                     1Department of Neuroscience, Reproductive Sciences
                     PO027                                                           and Dentistry, School of Medicine, University of Naples
                                                                                     Federico II, Naples, Italy; 2Department of Advanced
                     ANTIXA DIRECT ORAL ANTICOAGULANTS IN                            Biomedical Sciences, Federico II University Hospital,
                     ATYPICAL SITE VEIN THROMBOSIS: A SINGLE                         Naples, Italy; 3Division of Cardiology “Villa dei Fiori”
                     CENTRE EXPERIENCE                                               Hospital, Acerra, Naples, Italy; 4Zena and Michael
                                                                                     AWiener, Cardiovascular Institute/Marie-Josee and
                     A. Vitale2, A. Guida1, A. De Sena1, P. Tirelli1, N. Vitelli2,   Henry RKravis Center for Cardiovascular Health, Icahn
                     M. Bottiglieri2, A. Cardamone2, O. Iacono2, G. Landi2,          School of Medicine at Mount Sinai, New York, USA;
                                                                                     5Department of Clinical Medicine and Surgery, Federico
                     L. Palmisano2, M. Lugarà1, M.G. Coppola1,
                     P. Madonna1                                                     II University Hospital, Naples, Italy; 6Departmenttof
                                                                                     Advanced Biomedical Science, Division of Adult and
                     1UOC   Medicina Interna Ospedale del Mare ASL Napoli            Pediatric Cardiac Surgery, University of Naples




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Federico II, Naples, Italy; 7Pathology Unit, Department       (MCA) (Brain Sparing). From admission to the hospi-
                    of Public Health, School of Medicine, University of           tal, she underwent daily computerized-cardiotocogra-
                    Naples Federico II, Naples, Italy                             phy (c-CTG). Foetal Doppler velocimetry in UA and
                                                                                  MCA remained stable until 34 weeks. At 34 weeks,
                    Background: Antiphospholipid syndrome (APS) is a              patient developed gestational hypertension that was
                    systemic autoimmune disorder with acquired hyperco-           controlled with nifedipine (20 mg every 8 hours). At
                    agulability, characterized by venous, arterial and            34+3 weeks an emergency caesarean section was per-
                    microvascular thrombosis and increased pregnancy              formed. A female new born weighed 1290 grams was
                    morbidity, in the presence of persistently positive           born. The new born was admitted to Neonatal Intensive
                    antiphospholipid antibodies. Cardiac involvement may          Care Unit and she did not require ventilatory support.
                    be present as heart valve disease, affecting approxi-         Already after a week, she reached the weight of 1500
                    mately a third of patients, or, less frequently, as intrac-   grams and started oral feeding. She has been dis-
                    ardial thrombosis, nonbacterial thrombotic endocardi-         charged after two months. Currently enjoys good
                    tis (i.e. Libman-Sacks endocarditis), pulmonary hyper-        health. Conclusions: Intracardiac thrombosis or non-
                    tension, right or left ventricular dysfunction, micro-        bacterial thrombotic endocarditis is a rare complication
                    vascular thrombosis, coronary artery or micro-vascular        of APS. A correct management of pregnancy in patients
                    disease with overt or silent clinical presentation. Case      with this syndrome is fundamental: a multidisciplinary
                    Report: We report a case of Libman-Sacks tricuspid            approach should be required.
                    valve endocarditis in a 38-year-old pregnant woman at




                                                                                                         ly
                    15 weeks gestation with previously unknown primary
                    APS. During a routine cardiac examination and




                                                                                                  on
                                                                                  PO029
                    echocardiography, performed for a previous episode of
                    pleuro-pericarditis, a large mobile mass with irregular
                    edges was found at the level of the tricuspid valve.          INTRINSIC ALTERATION AND INFLAMMATORY




                                                                                            e
                    Three main differential diagnoses for intramyocardial         MICROENVIRONMENT AFFECT PROLIFERATION
                    mass were examined: malignancy, infective endocardi-
                    tis, nonbacterial thrombotic endocarditis. Cardiac mag-
                    netic resonance imaging (CMR) with contrast has
                                                                                     us
                                                                                  CAPABILITY OF CD8 T CELLS IN A MOUSE
                                                                                  MODEL OF HEMOPHILIA A

                                                                                  P.E. Di Simone, V. Kalandadze, A. Follenzi, C. Borsotti
                                                                                  al
                    raised the suspicion of a thrombus. The patient was
                    urgently admitted at the Cardiac Intensive Care Unit of
                                                                          ci

                    the Federico II University Hospital, and anticoagulant        Department of Health Sciences, Histology laboratory,
                    and antiplatelet therapy (Enoxaparin sodium at a dose         Università del Piemonte Orientale, Novara, Italy
                                                                    er


                    of 6000 IU every 12 hours and Acetylsalicylic Acid
                    100 mg daily) was started. Laboratory examination             Background and Aims: Hemophilia A (HA) is a reces-
                                                              m



                    showed prolonged partial thromboplastin time (PTT):           sive X-linked bleeding disorder caused by the lack of
                    63.6 seconds. Anticardiolipin antibodies IgG and anti-        coagulation factor VIII (FVIII). One of the main com-
                                                  om




                    β2 glycoprotein-1 IgG tested positive and they were           plications of the replacement therapy is the appearance
                    respectively 63 U/mL (cut off value <20 U/mL) and             of specific anti-FVIII antibodies (inhibitors) due to the
                    200 U/mL (cut off value <20 U/mL) with lupus antico-          B and T cells activation upon FVIII administration in
                                               -c




                    agulant (LA) positive. The laboratory examination and         hemophilic patients. Instead, little is known about any
                    the medical history induced to a diagnosis of primary         intrinsic T cell alteration especially prior to any FVIII
                                       on




                    APS, pending confirmation of the laboratory results at        exposure. Moreover, recently it has been reported the
                    12 weeks. Despite adequate anticoagulation for three          presence of pro-inflammatory cytokines in naïve
                    weeks, guided by the anti–factor Xa assays for dose           FVIII-/- mice associated to subclinical bleedings and in
                                  N




                    adjustment, transthoracic three-dimensional echocar-          HA previously untreated patients (PUPs) compare to
                    diography showed the persistence of the vegetation            healthy controls, suggesting that there could be a local
                    attached to the tricuspid valve, without changes in its       environment predisposing them to inhibitor formation.
                    size. A multidisciplinary consultation with obstetri-         Additionally, our lab observed in an hemophilic murine
                    cians, cardiologists, and cardiac surgeons was                model a slight decrease in number of HA splenic CD8
                    required. The patient decided not to terminate the preg-      T cells compare to wild type (wt) age- and sex-matched
                    nancy despite the risk to her health and to undergo car-      mice. In order to study the activation threshold of HA
                    diac surgery during pregnancy. Histological examina-          CD8 T cells we sought to investigate in vitro and in
                    tion confirmed the presence of nonbacterial thrombotic        vivo their proliferation capability. Methods: Splenic
                    endocarditis. Weekly obstetric controls were per-             CD8 T cells from HA and wt Balb/c mice were isolated
                    formed after surgery to verify foetal well-being.             and labelled with a fluorescent proliferation dye. An in
                    Pregnancy was complicated by foetal growth restric-           vitro assay was performed by anti-CD3/CD28±IL2
                    tion. At 28 weeks of gestation, foetal weight was below       stimulation while the in vivo test was run by adoptively
                    5th centile, with normal amniotic fluid and normal            transferring the CD8 T cells into immunodeficient HA
                    foetal Doppler velocimetry. At 32 weeks of gestation, a       NSG or NSG recipient mice. Results: HA CD8 T cells
                    PI of Umbilical Artery (UA) above 95th centile was            showed higher expansion compare to their wt counter-
                    found, with normal Doppler velocimetry of Ductus              parts. This difference was evident by the greater num-
                    Venosus (DV) and reduce PI in Middle Cerebral Artery          ber of HA CD8 T cells recovered after 72 hours of anti-




                                                               [Hematology Reports 2021; 13:s3]                                               [page 99]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 100



         Poster




                    CD3/CD28 in vitro stimulation (18410±3920 HA ver-             Genova, Italy; 4Fondazione IRCCS Ca’ Granda
                    sus [vs] 10298±1968 wt, p=0,001). On the other hand,          Ospedale Maggiore Policlinico, Angelo Bianchi
                    IL2 addition in the culture medium eliminated the HA          Bonomi, Hemophilia and Thrombosis Center, and
                    CD8 T cells proliferative advantage, suggesting a dif-        Università degli Studi di Milano, Department of
                    ferent intrinsic threshold of activation. Similarly, 72       Pathophysiology and Transplantation, Milan, Italy;
                    hours after the adoptive transfer, the splenic HA CD8 T       5Haemophilia and Transfusion Centre, “Bufalini”

                    cells recovery from the HA NSG recipient mice was 4-          Hospital, Cesena, Italy; 6Regional Reference Centre for
                    fold higher (425350±95312) than the wt CD8 T cells            Bleeding and Thrombotic Disorders, “Città della
                    recovery from NSG animals (91936±31319).                      Salute e della Scienza” University Hospital, Turin,
                    Surprisingly, tracking the number of cell divisions we        Italy; 7Haemophilia Regional Reference Centre,
                    did not only found more HA than wt CD8 T cells in             Haematology Unit, Policlinico “GRodolico” - SMarco
                    each cell cycle/division but also a different prolifera-      Hospital, Catania, Italy; 8Haemorrhagic and
                    tion kinetic with the highest number of proliferating         Thrombotic Disease Service, Area of Haematological
                    cells in the fourth and fifth cycle for HA cells and third    and Oncological Sciences, IRCCS Fondazione
                    and fourth ones for wt. In addition, the role played by       Policlinico Universitario “AGemelli”, Rome, Italy;
                                                                                  9Haemostasis and Thrombosis Service, Ospedale Civile
                    the host microenvironment was investigated by inject-
                    ing HA CD8 T cells into non-hemophilic NSG and wt             dell’Annunziata, Cosenza, Italy; 10“BTrambusti”
                    CD8 T cells into HA NSG: HA CD8 T cells recovery              General and Specialized Pediatrics Unit, “Giovanni
                    was higher from HA NSG than from NSG mice                     XXIII” Hospital, University of Bari, Italy;




                                                                                                          ly
                                                                                  11Haemophilia Centre, Internal and Cardiovascular
                    (425350±95312 vs 229419±46944 respectively,
                    p=0,11) while wt CD8 T cells number were low from             Medicine, “Santa Maria della Misericordia”




                                                                                                  on
                    both HA NSG and NSG (117324±38776 vs                          University Hospital, Perugia, Italy; 12Department of
                    91936±31319 respectively, p=0,68). Therefore, a com-          Clinical and Experimental Medicine, University of
                    bination of intrinsic and extrinsic factors might con-        Foggia, Foggia, Italy; 13Haemophilia and Thrombosis




                                                                                             e
                    tribute to reduce the threshold of proliferative activa-      Centre, University Hospital, Bari, Italy; 14Regional
                    tion in hemophilic CD8 T cells. Conclusions: Our pre-
                    liminary results showed a higher proliferation capabil-
                    ity of HA than wt CD8 T cells, supporting the hypoth-
                                                                                      us
                                                                                  Reference Centre for Inherited Bleeding and
                                                                                  Thrombotic Disorders, Transfusion Medicine, “Regina
                                                                                  Margherita” Children Hospital, Turin, Italy;
                                                                                  15Hematology Unit, Castelfranco Veneto Regional
                                                                                  al
                    esis of a possible intrinsic HA T cell alteration and/or
                    the influence of a tissue pro-inflammatory microenvi-         Hospital, Castelfranco Veneto, Italy; 16Hematology,
                                                                          ci

                    ronment. Our preliminary observations, which require          University Hospital Policlinico Umberto I, Rome, Italy;
                                                                                  17Regional Reference Center for Hemophilia and
                    further confirmations, could have an impact on the
                                                                    er


                    development of new strategies for preventing or eradi-        Coagulation Diseases, Azienda Ospedaliera Pugliese-
                    cating inhibitors in HA patients since it is possible that    Ciaccio, Catanzaro, Italy; 18Haematology Unit,
                                                              m



                    they already hold a pro-inflammatory signature prior to       Thrombosis and Haemostasis Reference Regional
                    any FVIII treatment.                                          Center, University of Palermo, Palermo, Italy;
                                                  om




                                                                                  19Hemophilia Centre, Grande Ospedale Metropolitano,

                                                                                  Reggio Calabria, Italy; 20Haemostasis and Thrombosis
                      PO030                                                       Unit, Haematology Department, San Bortolo Hospital,
                                               -c




                                                                                  Vicenza, Italy; 21SSD Congenital haemorrhagic
                                                                                  diseases, IRCCS Azienda Ospedaliero-Universitaria di
                    IDEAL STUDY: A REAL-WORLD ASSESSMENT OF
                                       on




                                                                                  Bologna, Bologna, Italy; 22Hemophilia and Thrombosis
                    PATTERN OF USE AND CLINICAL OUTCOMES                          Center, Hematology Unit, “Ospedale del Mare”
                    WITH RECOMBINANT FACTOR IX ALBUMIN                            Hospital, Naples, Italy; 23Haemophilia Centre,
                                  N




                    FUSION PROTEIN (RIX-FP) IN PATIENTS WITH                      Department of Medicine, University Hospital of Padua,
                    HAEMOPHILIA B IN ITALY                                        Padua, Italy; 24CSL Behring, Milan, Italy;
                                                                                  25Hippocrates Research, Genova, Italy; 26Department
                    G. Castaman1, A. Tagliaferri2, A.C. Molinari3,                of Health Sciences, University of Genoa, Italy
                    F. Peyvandi4, A. Coppola2, F. Demartis1, C. Biasoli5,
                    A. Borchiellini6, D. Cultrera7, R. De Cristofaro8,            Background and Aims: Factor IX replacement therapy
                    F. Daniele9, P. Giordano10, E. Marchesini11,                  is used for treatment and prophylaxis of bleeding in
                    M. Margaglione12, R. Marino13, B. Pollio14,                   hemophilia B. rIX-FP is a long-acting albumin-fusion
                    P. Radossi15, C. Santoro16, R.C. Santoro17, S. Siragusa18,    protein, which, in clinical studies, has demonstrated pro-
                    G. Sottilotta19, A. Tosetto20, L. Valdrè21, M.R. Villa22,     longed dosing intervals up to 21 days for routine prophy-
                    E. Zanon23, A. Loraschi24, I. Schiavetti25,26, D. Vaccari26   laxis, providing therapeutic benefit. The aim of the
                                                                                  IDEAL study was to describe dosing frequency and con-
                    1Center  for Bleeding Disorders and Coagulation,              sumption (primary endpoint), efficacy and safety of rIX-
                    Department of Oncology, Careggi University Hospital,          FP treatment during routine clinical practice in Italy.
                    Florence, Italy; 2Regional Reference Centre for               Methods: Patients with moderate/severe hemophilia B
                    Inherited Bleeding Disorders, University Hospital of          on prophylaxis with rIX-FP for ≥6 months, providing
                    Parma, Italy; 3Regional Reference Center for                  written informed consent, were enrolled in this observa-
                    Hemorrhagic Diseases, GGaslini Children’s Hospital,           tional study from October 2017 to February 2019 and




         [page 100]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 101



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    followed-up for two years. Descriptive analysis included     PO031
                    prospective and retrospective data (12 months prior to
                    switching to rIX-FP). The study was approved by com-
                    petent Ethics Committees. Results: Data were collected       IMPACT OF COVID-19 ON PEOPLE WITH
                    from 59 male patients (median age 30.1 ±16.20 years,         HEMOPHILIA: NEEDS AND SHORTINGCOMINGS
                    75% ≥18 years) enrolled by 23 hemophilia treating cen-       DUE TO PANDEMIC
                    ters. Of them, 50 (84.7%) were on prophylaxis during the
                    entire observation period and completed the study. The       G. Sottilotta1, D. Messina2, A. Buzzi3
                    infusion frequency changed from every 2-3 times/week         1Centro Emofilia – Servizio Emostasi e Trombosi, Grande
                    in 86.0% of patients with previous treatment to longer
                    than once a week in 84.0% of patients receiving rIX-FP       Ospedale Metropolitano di Reggio Calabria, Italy;
                                                                                 2AIMeF, Italy; 3Fondazione Paracelso, Milan, Italy
                    at the second-year follow-up. In particular, 54% of the
                    patients were on prophylaxis regimens every 8 to 10
                    days, 24.0% every 11 to 14 days and 6% every 15 to 21        Background and Aims: The large allocation of human
                    days. The prescribed dose remained quite stable during       and economic resources to face the pandemic and the
                    the entire 2-years of follow up (mean value ranging from     lockdown mobility restrictions have limited the patients
                    43.4 IU/kg to 46.0 IU/kg), as well as the frequency of the   access to in-person care at Hemophilia Treatment
                    infusions. At the end of the study, the annualized total     Centres (HTC) engaging in adopting new methods to
                    dose decreased from 4098.2±1340.10 IU/kg with previ-         ensure their continuity of care. Although the community




                                                                                                          ly
                    ous FIX, to 1562.6±433.23 IU/kg of rIX-FP. With an           of persons with haemophilia (PWH) has shown to be
                                                                                 resilient and adaptable, a large proportion of adults have




                                                                                                  on
                    annual number of infusions decreased of about 70%,
                    mean FIX activity trough level increased from 3.8% to        faced financial struggles due to job losses or suspensions.
                    14.4%. (Table 1). Median Annualized Bleeding Rate of         These multiple challenges have likely had a negative
                    0.00 was achieved across all prophylaxis regimens. Of        impact not only on PWH’s physical health but also on




                                                                                            e
                    note, the max ABR value decreased from 12 with the pre-      their psycho-social well-being7. An online survey was
                    vious treatment, to 7.7 with rIX-FP. Subjects with zero
                    bleedings increased from 66.0 % to 78.0% with rIX-FP.
                    In the 2-year follow-up, the rate of patients with chronic
                                                                                     us
                                                                                 conducted in 2021 with the aim to evaluate the emotions
                                                                                 which have affected the adult haemophiliacs daily life,
                                                                                 from different countries. Methods: An 18-item anony-
                                                                                 mous online international survey evaluated the feelings
                                                                                 al
                    joint pain decreased from 44.0% with previous treatment
                    to 16.0%, while that of patients with target joints from     of adult hemophiliacs, representing a variety of cultural
                                                                         ci

                    30.0% to 8.0%. No safety concerns were identified, no        and economic settings, and different health care systems,
                    patient developed inhibitors. Conclusions: In this real-     during the first year of pandemic and what behaviours
                                                                   er


                    world setting, approximately 80% of patients receiving       have been adopted to face the socio-economic restric-
                    rIX-FP are maintained on prophylaxis regimens with           tions including lockdown, social distancing and self-iso-
                                                             m



                    infusion frequency less than once a week, showing high-      lation, as well as the limited access to HCT. Results: The
                    er FIX levels, increased number of subjects with zero        participating patients were 204 from 24 different coun-
                                                 om




                    bleedings, zero median ABR, decreased chronic Joint          tries. The demographic and clinical characteristics are
                    pain and target joints and notably reduced FIX consump-      summarized in Table 1. Regarding the questions about
                    tion vs. previous treatment.                                 emotions and feelings in relation to the pandemic, the
                                              -c




                                                                                 sample of people interviewed replied as follows: 23.1%
                    Table 1.                                                     “not at all”, 45.6% “a little bit”, 16.6% “moderately”,
                                       on




                                                                                 10.8% “quite a bit”, 3.9% “extremely”. To question “Did
                                                                                 you find it difficult to manage the relationships with the
                                                                                 professionals?” the people interviewed replied: 48.5%
                                  N




                                                                                 that nothing had changed, 16.2% found no difficulty,
                                                                                 19.6% answered that it was difficult, 7.3% that it was
                                                                                 very difficult, 8.4% gave other answers. To question
                                                                                 about which aspect of the current situation has had the
                                                                                 greatest impact on their own life, the sample of people
                                                                                 interviewed replied as follows: 10.8% had “difficulties
                                                                                 related to the condition”, 8.3% “difficulties related to the
                                                                                 job, 11.8% “the obligation to stay at home”, 25.3%
                                                                                 “adapting to new habits and lifestyles”, 10.3% “obliga-
                                                                                 tion to use face masks, keep the distance and wash hands
                                                                                 frequently”, 12.7% “fear for my own and other people’s
                                                                                 health “, for 6.4% it was the economic impact, 5.4%
                                                                                 replied “the restrictions when traveling”, 7.8% “inability
                                                                                 to meet the loved ones”. To question about whether and
                                                                                 what kind of help the interviewee needed, 21.1% replied
                                                                                 that “the support of friends or family, even if virtual”,
                                                                                 47.1% replied that they needed more information; 28.4%
                                                                                 reported “psychological help”. 7.8% said they needed




                                                              [Hematology Reports 2021; 13:s3]                                              [page 101]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 102



         Poster




                    help in managing daily expenses. 15.7% reported need-           performed. Inhibitor patients: 4. Before switching to emi-
                    ing a greater presence of the HCT, 6.4% by the GP, and          cizumab: 3 on bypassing agents on demand; one patient
                    25.5% said they did not need help. About the check-ups          on rFVIII prophylaxis developed a high titer inhibitor.
                    and the planned activities at their HCT had been affected       Two patients received ITI. All patients switched to emi-
                    by the pandemic: for 40.2% there was no change; for             cizumab for a better control of the disease. No sponta-
                    24% there was a reduction in all activities; 27.4% were         neous bleeding were reported. A traumatic fracture of the
                    delayed only in programmable activities such as check-          homerus occurred, treated with 4 doses of rFVIIa
                    ups; 4.9% answered that the pandemic caused a post-             (90mcg/kg/dose) without any complication. One patient
                    ponement of the switch to a new drug, while 3.5% gave           underwent a colonoscopy without additional prophylaxis
                    other responses. Conclusions: The survey revealed that          (no biopsies performed). Two patients underwent dental
                    in a condition of social isolation, it is the experience of a   extraction: just one received prophylaxis with tranexamic
                    sense of uncertainty that prevails, which, although it is       acid; no hemorrhagic complications were observed.
                    not possible to eliminate completely, can however be            Conclusions: We confirm the efficacy and safety of emi-
                    managed, if continuity in relationships and support is          cizumab prophylaxis. No spontaneous bleedings
                    guaranteed even at a distance.                                  occurred. Two traumatic bleedings were easily managed
                                                                                    with rFVIIa or FVIII concentrate. Dental extractions in
                    Table 1.                                                        inhibitor patients were performed without rFVIIa thera-
                                                                                    py. No serious adverse events observed. The different
                                                                                    median age and follow-up between inhibitor and non-




                                                                                                            ly
                                                                                    inhibitor patients depends on timelines of regulatory
                                                                                    indications for prescription and previous participation in




                                                                                                     on
                                                                                    clinical studies. We observed an improvement in quality
                                                                                    of life of patients and their families.




                                                                                               e
                                                                                    Table 1.

                                                                                        us
                                                                                    al
                                                                            ci
                                                                     er


                      PO032
                                                               m



                    EMICIZUMAB PROPHYLAXIS: EXPERIENCE OF A
                    SINGLE CENTER
                                                   om




                    G. Maestrini, E. Baldacci, A. Ferretti, R. Mormile,
                    S. Olivieri, A. Pallotta, F. Viola, A. Chistolini,
                                                -c




                    C. Santoro
                                        on




                    Dipartimento di Medicina Traslazionale e di Precisione,         PO033
                    Istituto di Ematologia, Università “La Sapienza”, Roma,
                    Italy
                                   N




                                                                                    MANAGEMENT OF PATIENTS WITH BLEEDING
                    Background and Aims: The use of emicizumab as pro-              DISORDERS AND EDUCATIONAL NEEDS
                    phylaxis for severe HA, with or without inhibitor, is           REGARDING HEMOPHILIA A IN 23 ITALIAN
                    increasing in routine clinical practice. The aim of the         EMERGENCY DEPARTMENTS – RESULTS FROM A
                    study is to describe our center experience with emi-            SURVEY CONDUCTED IN PIEDMONT
                    cizumab prophylaxis in severe hemophilia A (HA)
                    patients without and with inhibitors. Methods: All the          B. Pollio1, I. Ricca1, C. Dainese2, F. Valeri2,
                    participants signed an informed consent; data were col-         A. Valpreda3, C. Linari3, A. Borchiellini2
                    lected from clinical charts. Results: 10 severe HA
                    patients received emicizumab prophylaxis (Table 1). We          1Centro di Riferimento Regionale Malattie Emorragiche
                    observed 1 drug-related adverse event, a relapsing orti-        e Trombotiche Ereditarie in Età Pediatrica, AOU Città
                    carioid reaction at the injection site. Non-inhibitor           della Salute e della Scienza -Ospedale Infantile Regina
                    patients: 6. Four patients were on rFVIII prophylaxis, 1        Margherita di Torino; 2Centro di Riferimento Regionale
                    was on demand therapy, 1 was never treated. Reason for          Malattie Trombotiche ed Emorragiche dell’aAdulto,
                    starting emicizumab: need to facilitate the therapy             Ematologia Universitaria, AOU Città della Salute e
                    administration in young children. Just one traumatic            della Scienza di Torino; 3Struttura Complessa di
                    tongue lesion occurred which was treated with one dose          Biochimica Clinica, AOUCittà della Salute e della
                    of rFVIII. None surgical or invasive procedures were            Scienza di Torino, Italy




         [page 102]                                         [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 103



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Background: Haemophilia and von Willebrand dis-               Background: Haemophilia A is a recessive, X-linked
                    ease are the bleeding disorders most frequently               inherited blood disorder caused by a deficiency of coag-
                    encountered in the emergency department (ED), that            ulation factor VIII (FVIII). The severity of haemophilia
                    are often the first point of contact for patients. evidence   is inversely related to the level of functional FVIII and
                    suggests that management in the ED is currently sub-          is categorized into mild, moderate, and severe disease,
                    optimal, mainly because the physicians have few               as follows: severe (factor level <1% of normal), moder-
                    opportunities to deal with this kind of patients. We car-     ate (level 1–5% of normal) and mild (level >5%).
                    ried out a survey to investigate the management of            Although haemophilia patients appear to have lower
                    patients with haemophilia A in Emergency                      mortality due to vascular diseases compared to the gen-
                    Departments (EDs), and to understand the training             eral population, the prevalence of cardiovascular risk
                    needs of the involved physicians. Methods: Overall, in        factors (i.e. arterial hypertension) in these patients is
                    Piedmont Region there are 32 EDs, and considering             similar to the prevalence in the general population.
                    that our survey was conducted on 21 physicians work-          Nonetheless, haemophilic patients have the same degree
                    ing in 23 Emergency Departments (EDs), this number            of atherosclerosis as the general population and the inci-
                    is representative of the Region’s reality. The interviews     dence of vascular thrombosis in these patients is increas-
                    were conducted through face-to-face meetings, includ-         ing, as life expectancy of these patients approaches
                    ing general aspects regarding the clinical characteris-       nowadays that of the general population. We report on a
                    tics and the management of patients, and self-evalua-         mesenteric arterial thrombosis in a mild haemophilia A
                    tion of knowledge and interest in receiving information       patient, with concomitant acquired and inherited throm-




                                                                                                          ly
                    about the disease. Results: In 2019, 131 patients with        bophilia. Case Report: A 51-year-old male patient was
                    haemophilia A were admitted (108 adults, 23 paedi-




                                                                                                   on
                                                                                  admitted because of severe abdominal pain and diarrhea.
                    atric). The best-known and most widely available and          Computer tomography (CT) revealed thrombotic occlu-
                    used treatments were plasma derivatives, followed by          sion of the superior mesenteric artery. The clinical histo-
                    first- and second-generation recombinant FVIII. More          ry of the patient included arterial hypertension, dyslipi-




                                                                                             e
                    recent recombinant and bypassing agents were less             demia, and smoking habit. Laboratory examination
                    known. Half of the interviewees considered their
                    knowledge of bleeding disorders in general and
                    haemophilia in particular to be “basic”, and only one
                                                                                      us
                                                                                  showed a prolonged activated partial thromboplastin
                                                                                  time (aPTT): 41,4sec (ratio: 1.4), with normal prothrom-
                                                                                  bin time (PT). Coagulation factor IX (FIX), FXI, and
                    third defined it as “good”; however, 86% expressed
                                                                                  al
                                                                                  FXII, and fibrinogen were normal. The FVIII level was
                    great interest in receiving information about the topic       33%. The patient reported the presence of heamophilia
                                                                          ci

                    (Figure 1). Conclusions: The survey confirms the              A in a family member. Molecular analysis showed the
                    needs related to the clinical management of rare inher-       presence, in hemizygosis, of the sequence variant
                                                                    er


                    ited clotting disorders in EDs. The physicians involved       c.1649G>A (p.Arg550His), in exon 11 of the F8 gene,
                    are keen to overcome this lack of knowledge, and prop-        present in other members of the same family. Lupus
                                                              m



                    er initiatives should be implemented.
                                                                                  anticoagulant was positive (1.76), anti-β2Glycoprotein I
                                                                                  antibodies IgG were positive (13672 GPL), anticardi-
                                                  om




                                                                                  olipin antibodies IgG were also present at high titres
                                                                                  (1487 GPL), suggesting a diagnosis of antiphospholipid
                                                                                  syndrome. The search for inherited thrombophilia
                                               -c




                                                                                  showed the presence of the FII G20210A variant. A re-
                                                                                  evaluation of antiphospholipid antibodies was planned
                                       on




                                                                                  at 3 months. The patient was treated with the association
                                                                                  of warfarin (target INR 2-3) and acetylsalicylic acid
                                                                                  (100 mg daily), without bleeding complications.
                                  N




                                                                                  Discussion: The management of adult haemophilic
                                                                                  patients with thrombotic events is often challenging
                    Figure 1.                                                     because of coexistence of the high bleeding risk and the
                                                                                  multiple co-morbidities and prothrombotic risk factors.
                                                                                  The levels of FVIII (mild hemophilia) and the coexis-
                    PO034                                                         tence of a pro-thrombotic state prevented bleeding com-
                                                                                  plications of the antithrombotic treatment in this patient.
                    MESENTERIC ARTERIAL THROMBOSIS IN A
                    PATIENT WITH HAEMOPHILIA A                                    PO035
                    A. Casoria1, M. Amitrano2, G. Antignani2, M. Nunziata2,
                    I. Puca2, F. Cannavacciuolo2, M. Mastroianni2,                PLASMA CONCENTRATION OF EMICIZUMAB IN
                    E. Cimino1, A. Tufano1                                        CHILDREN WITH HAEMOPHILIA A WITH AND
                                                                                  WITHOUT INHIBITORS
                    1Department   of Clinical Medicine and Surgery, Federico
                    II University Hospital, Naples, Italy; 2Internal Medicine     M. Leotta, A. Strangio, A. Ierardi, C. Spadafora,
                    Ward, San Giuseppe Moscati Hospital, Avellino, Italy          R.C. Santoro




                                                               [Hematology Reports 2021; 13:s3]                                             [page 103]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 104



         Poster




                    Hemostasis and Thrombosis Center, Azienda                    PO036
                    Ospedaliera Pugliese-Ciaccio, Catanzaro, Italy

                    Emicizumab is a bispecific, humanized, monoclonal            MILD HAEMOPHILIA A AND B: FROM BLEEDING
                    antibody mimicking the factor (F) VIII cofactor activi-      PHENOTYPE TO ILLNESS PERCEPTION
                    ty in mediating the generation of FXa by FIXa in
                    patients with hemophilia A (HA).. This subcutaneous          M. Mattana1, C. Rotolo1, R. Tomasello1, C. Cammarata1,
                    non-factor agent has been extensively approved for the       F. Romano1, A. Lucchesi2, G. Giordano3, S. Siragusa1,
                    prophylaxis of patients of HA patients with and with-        M. Napolitano1
                    out FVIII-inhibitors of all ages. Once weekly (QW)
                                                                                 1Department of Health Promotion, Mother and Child
                    administration of emicizumab increased plasma emi-
                    cizumab concentrations, and reached steady-state after       Care, Internal Medicine and Medical Specialties
                    4 weeks of weekly loading dose of 3 mg/kg body-              (ProMISE), University of Palermo, Palermo, Italy;
                                                                                 2IRCCS Istituto Romagnolo per lo studio dei tumori
                    weight followed by QW maintenance dose of 1.5mg/kg
                    bodyweight. Subsequently in patients without                 (IRST)”Dino Amadori”, Meldola (FC), Italy; 3Division
                    inhibitors, maintenance regimens were used which             of Internal Medicine, Hematology Service, Regional
                    included, as an alternative to the dosage of 1.5 mg / kg     Hospital “ACardarelli”, Campobasso, Italy
                    / week, dosages of 3 mg / kg every 2 weeks or 6 mg /
                    kg every 4 weeks. Maintenance doses allow,then, to           Background and Aims: Haemophilia is an X-linked




                                                                                                          ly
                    maintain constant plasma levels of emicizumab over           recessive disease, which mainly affects men. Disease
                    time and are not recommended changes in the dosage           severity depends on the plasma levels of factor VIII




                                                                                                  on
                    of the drug. Furthermore, no dosage adjustments are          (FVIII) in Haemophilia A (HA) or factor IX (FIX) in
                    required in relation at the age of the patient or any        Haemophilia (HB), mild haemophilia (MH) is defined by
                    comorbidities. Therefore, it is not necessary to deter-      FVIII or FIX plasma levels between 5% and 40%. MH is




                                                                                            e
                    mine the plasma concentrations of the drug over time,        often diagnosed accidentally due to excessive bleeding
                    unless there is suspicion of the formation of anti-drug
                    antibodies. Methods: We wanted to verify the plasma
                    concentrations of emicizumab and their variations in
                                                                                     us
                                                                                 secondary to trauma or surgery. Patients with MH do not
                                                                                 usually require prophylaxis and are treated on-demand,
                                                                                 they are less regular in follow-up visits and may under-
                    predetermined times (after the 4 loading doses, after 6      estimate their haemorrhagic risk. There is a paucity of
                                                                                 al
                    months, after 1 year and after 2 years) in 3 children        data focused on MH; clinical evidence and guidelines are
                                                                         ci

                    with hemophilia A. Two of the children, aged 8 and 6,        predominantly derived from severe haemophilia data.
                    respectively, suffering from haemophilia A with a high-      The current study aimed to describe clinical, laboratory
                                                                   er


                    titre inhibitor, were maintained at a dosage of 1.5 mg /     characteristics and quality of life and illness perception
                    kg / body weight. The third child, 3 years old, with         of a cohort of patients affected by MH; we have further-
                                                             m



                    severe haemophilia A without inhibitor, practiced            more evaluated differences, if any, between patients with
                    maintenance at a dosage of 3 mg / kg / body weight           FVIII:c and FIX:c levels below and above 10%.
                                                 om




                    every 2 weeks. Results: The median plasma concentra-         Methods: This retrospective study enrolled all consecu-
                    tion of emicizumab after loading doses was 52.3 μ /ml        tive patients affected by mild haemophilia A or B, evalu-
                    (50.5- 59.4). Dopo 6 mesi 60,5 μ /ml ( 48,7 - 84,4).         ated at our reference center from January 1, 2012, to
                    Dopo 12 mesi 59,7 μ /ml ( 51,2- 94). Dopo 24 mesi            November 30, 2021. Age, sex, FVIII and FIX levels,
                                              -c




                    65,6 μ /ml (42,3-89,5). The 24 month data is not avail-      spontaneous or provoked bleeding episodes were collect-
                    able for the patient without inhibitor. In our patients,
                                       on




                                                                                 ed. Patients were divided into two groups: A for patients
                    the median plasma concentration of emicizumab is             with factor levels less than or equal to 10%, B for patients
                    consistent with the literature data and remains constant     with factor levels higher than 10%. The Chi-square test
                                  N




                    over time, also confirming adherence to treatment. In a      was used for normally distributed continuous data analy-
                    single patient, with inhibitor, the values are consistent-   sis. A p-value <0.05 was considered statistically signifi-
                    ly higher (Table 1). Conclusions: We confirm that in         cant. Patients with MH filled also the SF36 and ED-5Q
                    our young patients, suffering from haemophilia A with        questionnaires, to assess health related quality of life and
                    and without inhibitor, the plasma levels of emicizumab       aspects of illness experience and perception and the
                    remain constant over time and always above the mini-         results expressed as a percentage. Results: Forty-two
                    mum levels reported in the literature.                       male patients with MH were analysed. The average age
                                                                                 was 42.6 +/- 23.4 years. Of these patients, 14 had FVIII
                    Table 1.
                                                                                 or FIX levels less than or equal to 10%; the remaining 28
                                                                                 had levels greater than 10%. Haemorrhagic manifesta-
                                                                                 tions and related therapies are shown in Figure 1.
                                                                                 Seventeen patients received short-term prophylaxis for
                                                                                 dental extraction and 12 for surgery. All the enrolled
                                                                                 patients were treated on demand except for two, under
                                                                                 continuous prophylaxis for recurrent bleeding. The
                                                                                 analysis revealed in-group A a statistically significant (p
                                                                                 <0.05) higher incidence of hemarthrosis and spontaneous
                                                                                 bleeding than in-group B. There were no statistically sig-




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 105



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    nificant differences regarding bleeding secondary to trau-   afterwards he was switched with Emicizumab prophy-
                    ma or surgery. SF36 and EQ5D were available for 20           laxis. In January 2022 the patient contracted a pau-
                    patients: 8 patients from group A and 12 from group B.       cisymptomatic Sars-CoV2 infection. In March 2022 the
                    SF36 questionnaire scored above average (50 SD +/-10)        patient presented acute onset of right hemiplegia and loss
                    in the area of physical function, and below average in the   of consciousness; trasported to the hospital, he presented
                    area of emotional aspects, with a 5/6 point difference.      low O2 saturation and Glasgow Coma Scale (GCS) 9 and
                    The impact of the disease on daily life is thus important,   was immediately intubated and moved into Intensive
                    and unrelated to Factor levels. The ED-5Q data are con-      Care Unit (ICU). The EEG performed in ICU showed
                    gruent with the SF36 data. Perceived health status, which    few slow and spiky/sharp potentials in the context of
                    mirror the state of disease awareness (Insight), was also    hypnic patterns with left prevalence. The brain MRI with
                    assessed, with results consistently above 70%.               angiographic sequences showed an uncertain thrombosis
                    Conclusions: The results of our analysis confirm that        of the right transverse sigmoid jugular venous axis: for
                    among patients with mild haemophilia, the most frequent      this reason he started antithrombotic prophylaxis with
                    bleeding events are secondary to trauma or surgery. Our      enoxaparine (100 UI/kg). After few hours the patient
                    analysis found that spontaneous bleeding and hemarthro-      recovered, thus he was extubated and when awaked he
                    sis are more common in patients with factor levels below     appeared in good general and neurological condition (no
                    10%, MH negatively impacts patients’ quality of life,        focal signs on neurological examination). He performed
                    especially for their emotional sphere.The haemophilia        another EEG where no epileptic anomalies were detected
                    community is called to act on patient education and          and another brain MRI showing an improvement of the




                                                                                                          ly
                    improvement of disease perception.                           previously described findings and, moreover, absence of




                                                                                                  on
                                                                                 recent onset areas of tissue suffering. Coagulation lab test
                                                                                 were normal, including d-dimer. According to these find-
                                                                                 ings, he discontinued antithrombotic therapy and started
                                                                                 valproic acid to prevent seizures. After 12 days patients




                                                                                            e
                                                                                 experienced a new and similar critical event with right
                                                                                     us
                                                                                 hemiplegia and loss of consciousness with a duration of
                                                                                 10 minutes without compromising respiratory activity, so
                                                                                 he didn’t need intubation. After the first critical episode,
                                                                                 al
                                                                                 emicizumab prophylaxis was stopped. Therefore, the
                                                                                 patient has undergone to a strict follow up (once a week)
                                                                         ci

                                                                                 by monitoring coagulation assays, including thrombotic
                                                                                 parameters (activated partial thromboplastin time, FVIII
                                                                   er


                                                                                 one-stage clotting assays, FVIII chromogenic assays, d-
                    Figure 1.                                                    dimer, anti-phospholipid antibodies). The patient has
                                                             m



                                                                                 resumed rFVIII-Fc prophylaxis (30 UI/kg) only after 25
                                                                                 days from last emicizumab infusion and continued with
                                                 om




                    PO037                                                        a weekly infusion for 4 weeks. After this period he
                                                                                 resumed regular frequency of prophylaxis three
                                                                                 times/week according to FVIII chromogenic assays with
                    A PARTICULAR CASE OF EPILEPSY IN A CHILD
                                              -c




                                                                                 a trough level of 1.9% according to pk assessment.
                    WITH HAEMOPHILIA AFTER SARS-COV2
                                                                                 Conclusions: The patient, despite having mild genetic
                                       on




                    INFECTION                                                    thrombophilia, did not experience thrombotic events dur-
                                                                                 ing prophylaxis with FVIII and also with emicizumab.
                    M.L. Tramontano1, G. Tesone1, M. Pandolfi2,                  We do not yet have elements to correlate with the previ-
                                  N




                    M. Della Corte2, E. Savastano2, M. Schiavulli1               ous SarsCoV2-infection but we suppose that this condi-
                    1Bleeding
                                                                                 tion, even if after about two months, may have played a
                               and Thrombosis Care Regional Reference            role in determining the onset of such serious episodes.
                    Center Santobono-Pausilipon Children’s Hospital, AORN,
                    Naples, Italy; 2Pediatric Neurology Unit Santobono-
                    Pausilipon Children’s Hospital, AORN, Naples, Italy          PO038
                    Background: Hemophilia A (HA) is an inherited bleed-
                    ing disorder that, if not properly treated, is associated    CLINICAL AND MOLECULAR CHARACTERIZATION
                    with debilitating joint damage due to recurrent              OF PATIENTS WITH INHERITED
                    hemarthroses as well as life-threatening bleeds including    DYSFIBRINOGENEMIA FOLLOWED IN A SINGLE
                    intracranial hemorrhage. The report describes an unusual     CENTER
                    case of epileptic seizure in a child affected by severe
                    hemophilia after sars Cov2 infection. Case Report: A 7       A. Strangio, A. Ierardi, M. Leotta, C. Spadafora,
                    years old male affected by Severe Hemophilia A.              R.C. Santoro
                    Heterozygous Prothrombin mutation (G20210A).
                    Autistic spectrum disorder. The patient started prophy-      Hemostasis and Thrombosis Center, Azienda
                    laxis with recombinant FVIII -Fc from 2016 to 2020,          Ospedaliera Pugliese-Ciaccio, Catanzaro, Italy




                                                              [Hematology Reports 2021; 13:s3]                                              [page 105]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 106



         Poster




                    Background and Aims: Fibrinogen is a 340 KDa gly-            Conclusions: Patients carrying the c.901 C>T mutation
                    coprotein that is encoded by three genes namely              in exon 8 in FGG gene had both thrombotic and hemor-
                    Fibrinogen Aα (FGA), Bβ (FGB), and γ (FGG).                  rhagic manifestations; patients with c.394_405 in exon
                    Inherited Dysfibrinogenemia is a rare functional fib-        4 in FGA gene only thrombotic episodes; patients with
                    rinogen disorder in which the fibrinogen protein is          c.160 T>G in the exon 2 in FGA gene only hemorrhag-
                    present but with a reduced function. The disorder is         es, all others were asymptomatic. Further studies to
                    characterized by a wide spectrum of clinical pheno-          assess      phenotype-genotype       correlation    of
                    types, ranging from asymptomatic to mild to severe           Dysfibrinogenemia are needed.
                    bleeding or thrombotic manifestations and recurrent
                    miscarriages. The aim of our study is to describe the
                    clinical phenotype in relation to genotype in our            PO039
                    cohort. Methods: We conducted a retrospective analy-
                    sis of 22 patients from 14 families with inherited dys-      OBSTETRICAL MANAGEMENT OF PATIENTS WITH
                    fibrinogenemia followed by our center. All patients          FACTOR XI DEFICIENCY: RETROSPECTIVE
                    were asked for consent to make a molecular diagnosis,
                                                                                 ANALYSIS OF A SINGLE CENTER
                    but consent was obtained from only 12 patients from 7
                    families. To assess the clinical phenotype, data was
                                                                                 A. Ierardi, A. Strangio, M. Leotta, C. Spadafora,
                    collected that included coagulation and molecular
                                                                                 R.C. Santoro
                    screening, gender, bleeding and thrombotic events.




                                                                                                        ly
                    Results: 8 patients are male and 14 female. 4 of the         Hemostasis and Thrombosis Center, Azienda Ospedaliera




                                                                                                 on
                    male and 8 female patients were asymptomatic. The            Pugliese-Ciaccio, Catanzaro, Italy
                    remaining male patients had experienced thrombotic
                    episodes, particularly ischemic hearth disease in 2          Background and Aims: Factor XI (FXI) deficiency
                    patient at the age of 42 and 48, respectively. Another




                                                                                           e
                                                                                 was first described in the 1950s. Managing patients
                    patient had an ischemic stroke at age 50 and the last
                    patient had idiopathic cerebral haemorrhage at age 40
                    and cerebral ischemia at age 65. 8 female were asymp-
                    tomatic. 2 women had post-partum haemorrhages and
                                                                                    us
                                                                                 with FXI deficiency presents several challenges,
                                                                                 including a lack of correlation of bleeding symptom
                                                                                 with FXI activity levelsThis study aimed to evaluated
                                                                                 pregnancy management of FXI deficiency patients.
                                                                                 al
                    two other women had hematomas after surgery, all of          Methods: We performed a retrospective chart review of
                    them before diagnosis. One woman had cerebral
                                                                         ci

                                                                                 patients with FXI deficiency who underwent childbirth
                    venous thrombosis and another had post-partum deep           or had miscarriages. Data on age, ethnicity, family or
                    vein thrombosis. Whole exone sequencing analysis
                                                                   er


                                                                                 personal history of bleeding, type of anesthesia and
                    which included testing of the fibrinogen genes FGA,          wheter used or not, any compications, and type and tim-
                    FGB and FGG, was performed in 12 patients. Five
                                                             m



                                                                                 ing of blood product or hemostatic agent administered
                    pathogenic mutations were found in the FGG gene:             in the peri-operative period were collected. Results: We
                    c.901 C>T in exon 1 in three cases; homozygosity for
                                                 om




                                                                                 identify 13 women with FXI deficiency who had 31
                    the mutation c.673 G>T in exon 7 in another case and         pregnancy ending in childbirth or miscarriage (21 vagi-
                    finally c.902 G>A in exon 8; in the latter case, a further   nal deliveries, 7 caesarean sections and 3 miscarriages).
                    associated mutation, of uncertain significance, was          At the moment of the event the mean age was 27 years
                                              -c




                    found c.401 G>A in exon 4. Seven pathogenic muta-            (20-37). 7/13 women, for a total of 16 pregnancies, did
                    tions were found in the FGA gene: c.394_405 in exon
                                       on




                                                                                 not know they had a FXI defect at the time of their first
                    4 in three cases; c.103 C>T in exon 2 in two cases and       pregnancy and one of them became aware of the defect
                    c.160 T>G in exon 2 in two cases (Table 1).                  after both of her deliveries. Their median level of FXI
                                  N




                                                                                 was 34% (25-37). One pregnancy resulted in miscar-
                    Tabella 1.                                                   riage, 2 caesarean sections under neuro-axial anesthe-
                                                                                 sia, 13 spontaneous vaginal deliveries. None of them
                                                                                 had a history of bleeding. One woman, with anamnesis
                                                                                 of heavy menstrual bleeding, had a first pregnancy with
                                                                                 partial placental abruption and concomitant haemor-
                                                                                 rhage at 20 weeks of gestation. On that occasion, a
                                                                                 Factor XI defect (1,4%) was diagnosed. The patient car-
                                                                                 ried the pregnancy to term with spontaneous delivery.
                                                                                 Subsequently the patient had 3 other pregnancies, two
                                                                                 completed with spontaneous birth, 1 abortion. In all
                                                                                 cases, FFP was administered, with no adverse events.
                                                                                 Other three women, without a history of bleeding and
                                                                                 with mild FXI defect (27%, 40% and 40% respectively)
                                                                                 had 6 pregnancies, 4 with vaginal delivery and two with
                                                                                 caesarean delivery (with general anesthesia). In none of
                                                                                 these cases were blood products or hemostatic agents
                                                                                 used. Finally, two women with severe FXI defect and




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 107



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    no history of bleeding (FXI 0.5 and 0.7 respectively)       ants at this location in patients with VWD. Methods:
                    had 5 pregnancies. The first had 3 caesarean deliveries     We identified 3 patients with p.Arg1205His, 2 with p.
                    and 1 miscarriage (Table 1). The second just one preg-      Arg1205Cys and 2 with p.Arg1205Leu. Laboratory
                    nancy carried out with spontaneous birth. In all cases      tests included FVIII, VWF:Ag and VWF:RCo at base-
                    both patients were prepared with FFP. No haemorrhagic       line and after a desmopressin test trial. The level of
                    or thrombotic events were reported. Conclusions: In         VWF propeptide was also assessed. Desmopressin was
                    our patients higher FXI levels correlate with a lower       administered subcutaneously at 0,3 µg/kg and
                    odds of bleeding. All of our patients with mild defects     FVIII/VWF measurements assayed 1 and 4 hours post-
                    have gone through childbirth without the need for           administration. Results: At baseline, patients with
                    replacement therapy. All the other patients were pre-       p.Arg1205His and p.Arg1205Leu showed very low
                    pared with the administration of FFP.                       FVIII and VWF measurements (FVIII:C 10-15 U/dL,
                                                                                VWF:Ag 7-11 U/dL and VWF:RCo 6-12 U/dL), while
                    Tabella 1.                                                  those with p.Arg1205Cys showed clearly higher levels
                                                                                (FVIII:C 34 – 45 U/dL, VWF:Ag 17 – 29 U/dL and
                                                                                VWF:RCo 21 – 28 U/dL). VWFpp/VWF:Ag ratio was
                                                                                >10 in all the patients with p.Arg1205His/Leu variants
                                                                                while those with p.Arg1205Cys had only a slight
                                                                                increase (VWFpp/VWF:Ag 2.9 – 4.4). After desmo-
                                                                                pressin, a similar pattern was observed in patients with




                                                                                                         ly
                                                                                p.Arg1205His/Leu variants, with rapid return to base-




                                                                                                 on
                                                                                line after 4 hours and an average VWF half-life of 2.1
                                                                                hours, while those with p.Arg1205Cys variant had lev-
                                                                                els 2-3 fold greater than baseline still after 4 hours, with
                                                                                a VWF half-life of 6-8 hours. Conclusions: Amino acid




                                                                                           e
                                                                                substitutions at p.Arg1205 location in VWF may induce
                                                                                    us
                                                                                variable decrease of VWF at baseline and different
                                                                                clearance after desmopressin. In particular, the substitu-
                                                                                tion with a cysteine residue seems to improve signifi-
                                                                                al
                                                                                cantly the laboratory phenotype, mitigating the
                                                                                increased clearance of mutant VWF, again highlighting
                                                                        ci

                                                                                the important role of cysteine residues in this multimer-
                                                                                ic molecule.
                                                                  er
                                                             m




                    PO040                                                       PO041
                                                 om




                    IMPACT ON VON WILLEBRAND FACTOR                             ORAL AND GYNAECOLOGICAL MANIFESTATIONS
                    SURVIVAL OF DIFFERENT AMINO ACID CHANGES                    IN TWO WOMEN WITH PLASMINOGEN
                                              -c




                    AT P.1205 IN D3 DOMAIN OF VWF MOLECULE                      DEFICIENCY

                                                                                L. Pieri1, S. Linari1, F. Demartis1, R. Fjerza1,
                                      on




                    L. Pieri1, S. Linari1, F. Demartis1, R. Fjerza1,
                    F. Salvianti1, M. Coppo1, M. Attanasio1,                    M.C. Di Stefano1, R. Marcucci2, G. Castaman1
                    M.C. Di Stefano1, E. Dalla Porta2, G. Castaman1
                                  N




                                                                                1Centre  for Bleeding Disorders and Coagulation,
                    1Centre  for Bleeding Disorders and Coagulation,            Department of Oncology, Azienda Ospedaliero-
                    Department of Oncology, Azienda Ospedaliero-                Universitaria Careggi, Florence, Italy; 2Department of
                    Universitaria Careggi, Florence, Italy; 2Department of      Clinical and Experimental Medicine, University of
                    Clinical and Experimental Medicine, University of           Florence, Florence, Italy
                    Florence, Florence, Italy
                                                                                Background: Plasminogen (PLG) deficiency is a rare
                    Background and Aims: Von Willebrand factor (VWF)            disorder associated with development of fibrin-rich
                    variant p.Arg1205His in D3 domain of VWF is associ-         pseudomembranes on mucosal surfaces, mainly affect-
                    ated with von Willebrand disease (VWD) Vicenza. This        ing the conjunctiva (ligneous conjunctivitis), gums (lig-
                    variant is by far that associated with the highest clear-   neous gingivitis), cervix and endometrium, ears,
                    ance of VWF, as demonstrated by very high ratio             nasopharynx, respiratory, gastrointestinal and geni-
                    between VWF propeptide and VWF antigen                      tourinary tracts. Abnormal wound healing and infertili-
                    (VWFpp/VWF:Ag) in circulation in several studies.           ty have also been reported. The estimated prevalence of
                    Little is known about the possible influence, if any, of    homozygous or compound heterozygotes is 1.6 affected
                    different amino acid substitution at the same location.     individuals per million population. Currently, there is
                    The aim of this study is to report the phenotypic           no specific replacement treatment approved for congen-
                    FVIII/VWF changes associated with amino acid vari-          ital PLG deficiency. We collected the medical history




                                                             [Hematology Reports 2021; 13:s3]                                              [page 107]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 108



         Poster




                    and describe therapeutic approach in two patients with       remains a matter of debate. Case Report: We report
                    PLG deficiency referred to our center. The laboratory        the case of a 79-year-old patient with type 2A VWD.
                    data included PLG activity measured with chromogenic         VWD was diagnosed at the age of 32 years because of
                    assay and genetic test. Case Report 1: A 35 year-old         prolonged bleedings after minor wounds. The labora-
                    woman with history of ligneous conjunctivitis and            tory evaluation showed FVIII:C 64.5% (normal range
                    recurrent pseudomembranes of the gingiva treated by          60-160%), VWF:Ag 37.3% (normal range 60-160%,)
                    surgical excisions with limited success. The gums            VWF:RCo 21.1% (normal range 60-160%) and
                    involvement led to multiple dental loss. The patient also    VWF:CB 19.1% (normal range 65-150%). In 2010,
                    presented vaginal and cervical pseudomembranes. The          the patient underwent a cholecystectomy and an exci-
                    PLG level was 19% (normal value 80-110%). The                sion of a benign lesion of the neck complicated by
                    genetic analysis showed a compound heterozygosity for        bleeding, which required hospitalization and infusion
                    aTrp616Cys and an Arg386* mutations. The                     of plasma-derived VWF/FVIII concentrate (Haemate
                    Trp616Cys is located at the catalytic domain of PLG          P®) for a few days. Moreover, the patient had a histo-
                    gene in exon 15, is reported with an allele frequency        ry of arterial hypertension, type 2 diabetes mellitus
                    0.001% without data on the clinical significance, while      and hypercholesterolemia. In 2015, following the
                    Arg368*, located in exon 10, results in a stop codon and     onset of exertion-induced chest pains, the patient was
                    it has not been reported previously. The patient per-        admitted to Padua University Hospital and resulted
                    formed a tooth extraction and implantology using pro-        positive to inducible myocardial ischemia. A coronary
                    phylaxis with fresh frozen plasma 25 mL/kg before the        angiography revealed a critical multi-vessels disease




                                                                                                        ly
                    procedure and after 24 hours (h), without complica-          involving the common arterial trunk, which required a
                    tions. The PLG level rised to 38% after 24 h and to 30%      multiple coronary artery bypass. Cardiac surgery was




                                                                                                 on
                    after 48 h. The patient is currently following an in vitro   successful, and no hemorrhagic complications
                    fertilization program for primary infertility. Case          occurred. After surgery, the patient started antiplatelet
                    Report 2: A 20 year-old woman presented with post            therapy with Cardioaspirin 100 mg. In 2016, under




                                                                                             e
                    coital bleeding and severe dysmenorrhea due to lig-          antiplatelet treatment, the patient underwent multiple
                    neous cervicitis and vaginal pseudomembranes. PLG
                    level was 16%. The genetic evaluation identified com-
                    pound heterozygosity for two novel unreported muta-
                                                                                     us
                                                                                 tooth extractions complicated by slight post-procedure
                                                                                 bleeding, despite the administration of a high VWF
                                                                                 levels FVIII concentrate (Fanhdi®). A pharmacokinet-
                                                                                 ics test with Wilfactin®, a highly purified plasma-
                                                                                 al
                    tions: a Lys38Glu and a Gly30Val substitutions, both in
                    critical domains for the protein function and estimated      derived VWF almost devoid of FVIII, was performed
                                                                         ci

                    as likely pathogenetic by the bioinformatic programs.        (Figure 1), and the patient started an on-demand regi-
                    Conclusions: PLG deficiency diagnosis is often               men because he refused prophylactic treatment. From
                                                                   er


                    delayed due to its rarity. Both patients underwent mul-      2016 to 2021, the patient suffered from recurrent epis-
                    tiple bioptic procedures before appropriate diagnosis.       taxis (up to 3-4 episodes a year); moreover, because of
                                                             m



                    The lack of specific therapy makes difficult the man-        the development of benign prostatic hypertrophy, and
                    agement of the clinical manifestations, that can be          urinary incontinence associated with many diverticu-
                                                 om




                    invalidating with consequent poor quality of life. The       la, a bladder catheter was placed. However, the patient
                    diagnosis can be easily performed by the PLG plasma          continued to refuse a prophylaxis regimen. In the last
                    level determination, once a clinical suspicion is made.      year, we recorded 4 episodes of epistaxis and 6
                                                                                 episodes of hematuria, of which at least 3 required
                                              -c




                    Genetic test could be useful for the diagnosis confirma-
                    tion and for a better understanding of the disorder, with    hospitalization. In October 2021, the patient agreed to
                                                                                 start prophylaxis with Wilfactin® at a dose of 2000 IU
                                       on




                    an assessment of structure/function relationships.
                                                                                 twice a week (less than 30 IU/kg). In the last six
                                                                                 months, only a single episode of bleeding (hematuria),
                                  N




                      PO042                                                      requiring an additional infusion of Wilfactin®, has
                                                                                 been reported. Conclusions: Prophylaxis with
                                                                                 Wilfactin® appear to be effective in preventing bleed-
                    EFFECTIVENESS OF PROPHYLAXIS USING                           ing in patients undergoing antiplatelet treatment.
                    WILFACTIN® IN PATIENTS WITH TYPE 2A VON
                    WILLEBRAND DISEASE AND ISCHEMIC HEART
                    DISEASE IN TREATMENT WITH ANTIPLATELET
                    THERAPY: A CASE REPORT

                    E. Zanon, C. Simion, E. Campello, L. Spiezia, P. Simioni

                    Haemophilia Center and General Internal Medicine,
                    Department of Medicine – DIMED, University-Hospital
                    of Padua Medical School, Padua, Italy

                    Backgound: Management of von Willebrand disease
                    (VWD) in patients with normal FVIII levels and car-
                    diovascular comorbidities treated with antiplatelets         Figure 1.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 109



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    PO043                                                         G.F. Guidetti1, I. Canobbio1, M. Brambilla3,
                                                                                  M. Camera3, S. Momi4, P. Gresele4, M. Torti1
                    TWO CASES OF HERMANSKY-PUDLAK                                 1Department
                    SYNDROME ASSOCIATED WITH NOVEL                                               of Biology and Biotechnology, University
                    MUTATIONS IN HPS3 AND HPS6 GENES                              of Pavia, Italy; 2University School for Advanced Studies
                                                                                  IUSS, Pavia, Italy; 3Centro Cardiologico Monzino
                    L. Pieri, M. Attanasio, M. Coppo, F. Salvianti, S. Linari,    IRCCS, Milano, Italy; 4Department of Medicine and
                    F. Demartis, R. Fjerza, M.C. Di Stefano, G. Castaman          Surgery, Division of Internal and Cardiovascular
                                                                                  Medicine, University of Perugia, Italy
                    Centre for Bleeding Disorders and Coagulation,
                    Department of Oncology, Azienda Ospedaliero-                  Background and Aim: Frailty represents a general state
                    Universitaria Careggi, Florence, Italy                        of excessive vulnerability frequently associated to aging
                                                                                  in humans, and significantly affects the quality of life.
                    Hermansky-Pudlak syndrome (HPS) is characterized by           Age-related frailty is more commonly associated to
                    oculocutaneous albinism, a bleeding diathesis, and, in        comorbidities such as obesity, diabetes, and Alzheimer’s
                    some individuals, pulmonary fibrosis, granulomatous           Disease (AD), but the underlying physiopathological
                    colitis, or immunodeficiency. HPS is a rare disorder with     mechanisms remain poorly understood (Chen et al., Clin
                    an estimated worldwide prevalence of 1-9 per 1,000,000
                                                                                  Interv Aging 2014; 9:433-441). It is generally recog-
                    individuals. The prevalence per subtype can differ due to
                                                                                  nized that a low grade chronic systemic inflammation




                                                                                                          ly
                    founder variants. The prevalence of HPS1-related HPS in
                    northwestern Puerto Rico is 1:1,800. A variable bleeding      plays a major role in the development of frailty (Soysal
                                                                                  et al., Ageing Res Rev 2016:31:1-8). Beside their role in




                                                                                                  on
                    diathesis associated with platelet function abnormalities
                    has been reported in some patients with the disorder. We      haemostasis and thrombosis, platelets act as true inflam-
                    report two novel cases of HPS in two children. The first      matory cells, and are implicated in some age-related
                    patient was born in 2019. At 3 months of age he devel-        pathologies, including AD (Catricalà et al., Immun




                                                                                             e
                    oped nystagmus with photophoby, no bleeding symptoms          Ageing 2012; 9:20). Therefore, platelets appear to be
                    were reported. Two novel heterozygous variants were
                    identified in HPS3 gene (c.1457G>A and c.1813G>T).
                    This gene codes for a protein of the complex BLOC-2
                                                                                      us
                                                                                  strategically located to link aging, frailty, and AD. We
                                                                                  have undertaken a longitudinal study to investigate the
                                                                                  correlation between platelet reactivity and the onset of
                    responsible for lysosome-associated organelle byosynthe-
                                                                                  al
                                                                                  frailty in a mouse model of AD. Materials and
                    sis. As expected, ADP and ATP release from platelets was      Methods: Transgenic APP23 mice, a consolidated
                    almost absent. The platelet count was normal and light
                                                                          ci

                                                                                  murine model for AD, were analysed during their lifes-
                    transmission aggregometry revealed only reduced aggre-        pan and experimental observations and analysis were
                                                                    er


                    gation after stimulation with epinephrine. The second         collected at 3, 9 and 18 months of age in comparison
                    patient was born in 2017 and soon after birth had nystag-     with control wild type (WT) of the same genetic back-
                    mus with foveal hypoplasia resulting in oculo-cutaneous
                                                              m



                                                                                  ground. Development of frailty was assessed according
                    albinism. No bleeding symptoms were reported.
                    Compound heterozygosity for two novel mutations has           to the methodology proposed by Whitehead et al. (J
                                                  om




                    been identified, both in the HPS6 gene which also codes       Gereontol 2014; 69:621-632) based on the non-invasive
                    for a protein of the complex BLOC-2 responsible for           evaluation of clinical signs of deterioration of phenotyp-
                    lysosome-associated organelle byosynthesis. The first         ic and physiological functions (i.e. tegument, ocular,
                                               -c




                    variant (c.1A>G) was identified in the asymptomatic           nasal respiratory systems, etc.). Platelet activation and
                    father and the second (c.210_211insGGGCC) in the              reactivity as well as platelet-leukocytes aggregates were
                                       on




                    asymptomatic mother. No release from platelet delta           evaluated by flow cytometry and lumiaggregometry.
                    granules was detected in the patient after stimulation with   Results: Clinical assessment of 31 specific clinical signs
                    ADP 2 and 10 microM and epinephrine, as a reduced ATP         of deterioration in mice allowed the calculation of a
                                  N




                    release after stimulation with arachidonic acid and colla-    cumulative frailty index that progressively increased
                    gen. Platelet count was normal, and light transmission        with age in both WT and APP23 mice demonstrating
                    aggregometry showed slight impaired aggregation after         that this approach is a valuable method to follow the
                    stimulation with ADP 2 microM and epinephrine. HPS is         progression of frailty in longitudinal studies on aged
                    a very heterogeneous disorder, although clinical oculo-       mice. In 18 months old mice, the frailty score for some,
                    cutaenous albinism is present in almost all patients. It is   but not all the specific evaluated parameters was higher
                    also confirmed that although low or absent delta granules     in APP23 mice compared to WT control animals of the
                    are detected, HPS3 and HPS6 variants are usually associ-
                    ated with no or mild bleeding tendency.                       same genotype, indicating that the development of AD is
                                                                                  associated to a stronger propensity to develop frailty.
                                                                                  Circulating platelets from aged APP23 mice displayed
                                                                                  an overall hyperactivation state compared to age-
                    PO044
                                                                                  matched controls, as demonstrated by a slightly higher
                                                                                  degree of integrin alpha(IIb)beta(3) activation and P-
                    BLOOD PLATELETS AT THE CROSSROAD                              selectin exposure. Aged APP23 platelets also revealed
                    BETWEEN ALZHEIMER’S DISEASE AND                               an evident hyperreactivity in terms of aggregation and
                    AGEING-RELATED FRAILTY                                        secretion upon stimulation with soluble agonists includ-
                                                                                  ing thrombin and convulxin. Importantly, the formation
                    M. Vismara1, S.M.G. Trivigno1,2, L. Galgano1,2,               of platelet-leukocyte aggregates was significantly




                                                               [Hematology Reports 2021; 13:s3]                                            [page 109]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 110



         Poster




                    enhanced in blood from aged APP23 mice compared to         the latters showed a significantly higher MFI
                    WT controls. Conclusion: These results show that age-      (1.11±0.44 in migraineurs vs 1.64±0.5 in ischemic PFO;
                    related frailty is more pronounced in a mouse model of     p<0.0001). Conversely TFPI expression was compara-
                    AD and is associated to hyperactivation and hyperreac-     ble in the 2 groups of patients thus resulting in a com-
                    tivy of circulating platelets. These observations point    parable platelet-associated TG. The number and the
                    toward a possible novel role of platelets in linking AD    MFI of ROSpos platelets was slightly increased in
                    with ageing-related frailty.                               ischemic PFO patients without migraine, suggesting a
                                                                               higher oxidative stress status compared to migraineurs.
                                                                               The total number of MVs was comparable between the
                      PO045                                                    2 groups of patients and the most abundant of them
                                                                               were from platelets. TF+-MVs were slightly higher in
                    INSIGHT INTO PROTHROMBOTIC PHENOTYPE                       ischemic patients (22.231±6671 in migraineurs vs
                                                                               23.182±5140 MVs/µl in ischemic PFO); PS+TF+-MVs
                    AND PLATELET ACTIVATION IN PATENT FORAMEN
                                                                               behaved similarly (4236±1556 in migraineurs vs
                    OVALE (PFO) PATIENTS WITH AND WITHOUT                      4777±125 MVs/µl in ischemic PFO). This resulted in a
                    MIGRAINE                                                   higher MV-associated TF-dependent TG in ischemic
                                                                               compared to PFO patients with migraine (p=0.0495).
                    P. Canzano1, D. Trabattoni1, M. Brambilla1,                Conclusions: This study shows that ischemic and
                    A. Becchetti1, M. Conti1, M. Camera1,2                     migraineur PFO patients share a similar platelet pro-




                                                                                                      ly
                    1Centro
                                                                               thrombotic phenotype. By contrast they differ in terms
                             Cardiologico Monzino IRCCS, Milan, Italy;




                                                                                               on
                    2Department
                                                                               of MV-associated TG, being that of ischemic patients
                                   of Pharmaceutical Sciences, Università      significantly higher than that of migraineurs.
                    degli Studi di Milano, Italy




                                                                                         e
                    Background and Aims: Patent foramen ovale (PFO) is         PO046
                    a interatrial communication with a prevalence of 25-
                    35%. Patients with cryptogenic stroke (CS) have a 2.3-
                    fold increased relative risk of having PFO, compared
                                                                                  us
                                                                               ADVANCES IN DIAGNOSIS OF HEREDITARY
                    with age-matched subjects with a clear-cause-stroke,       MACROTHROMBOCYTOPENIA: THE ROLE OF
                                                                               al
                    suggesting the PFO causative role of CS. Furthermore,      NEXT-GENERATION SEQUENCING
                                                                       ci

                    a strong relationship links PFO and migraine, a chronic
                    neurovascular disorder with a multifactorial aetiology.    P. De Bonis, A. De Laurenzo, E. Chinni, F. Cappucci,
                                                                  er


                    We recently showed that PFO patients with migraine         L. Fischetti, D. Colaizzo, E. Grandone, G.L. Tiscia
                    with aura (MHA) are characterized by a unique hyper-
                                                            m



                    active platelet state, featured by increased platelet      Unità di Emostasi e Trombosi, IRCCS Casa Sollievo
                    expression of Tissue Factor (TF) and phosphatidylser-      della Sofferenza, San Giovanni Rotondo (FG), Italy
                                                om




                    ine (PS). This procoagulant phenotype is completely
                    reverted by PFO closure with a significative regression    Background: About half of all cases of hereditary
                    of migraine symptoms, suggesting a biologic causative      macrothrombocytopenia are due to MYH9 (Myosin
                                                                               Heavy Chain 9) variations. However, as this condition
                                             -c




                    link between PFO and MHA. To date, there are no stud-
                    ies focused on platelet activation and procoagulant phe-   involves different stages of platelet development/differ-
                                      on




                    notype of CS PFO patients without migraine. Thus, aim      entiation and respective controlling genes can be
                    of this study was to perform a comparative analysis of     involved in the pathogenesis of the disease, the diagno-
                    platelet activation and prothrombotic potential between    sis may be challenging. On the other hand, molecular
                                  N




                    these patients and PFO patients with migraine.             diagnosis is needed, due to different management
                    Methods: 33 PFO with migraine and 27 ischemic PFO          strategies according the gene involved. Methods: We
                    without migraine patients on aspirin treatment were        describe two cases of macrothrombocytopenia
                    enrolled before PFO closure. Pselectinpos-, activated-     observed at Thrombosis and Hemostasis Centre of our
                    glycoproteinIIbIIIa (aGPIIbIIIa)pos-, TFpos-, Tissue       research Institute. Personal and family histories were
                    Factor Pathway inhibitor (TFPI) pos-, reactive oxygen      collected, with a focus on bleeding history. Laboratory
                    species (ROS)pos-platelets, platelet-leukocyte aggre-      investigations included complete blood count (BC) and
                    gates (PLA) and microvesicles (MVs) were evaluated         blood smear. Next-Generation Sequencing (NGS) was
                    by whole blood flow cytometry. Flow cytometry data         performed using an Illumina platform targeting a cus-
                    were reported as % of positive cells and levels of Mean    tom hereditary macrothrombocytopenia panel gene.
                    Fluorescence Intensity (MFI). TF-dependent Thrombin        Results: The first case here described is a 50-years-old
                    generation (TG) was assessed by Calibrated Automated       male with a previous diagnosis of immune thrombocy-
                    Thrombogram assay in the presence of TF and TFPI           topenia (ITP). He had no personal or family history of
                    neutralizing antibodies. Results: Expression of            spontaneous or post-traumatic bleedings (tooth extrac-
                    Pselectin and aGpIIb/IIIa, and the number of PLA, was      tions, minor surgery, etc.). He was diagnosed with ITP
                    similar in PFO patients with and without migraine. In      eight years before and treated with thrombopoietin
                    migraineurs, the number of TFpos platelets was compa-      (TPO) mimetics with partial response. Therefore, he
                    rable to ischemic PFO patients without migraine, but       underwent a comprehensive thrombocytopenia check




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 111



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    for ITP differential diagnosis. Blood smear examination       PO047
                    showed a platelets diameter of 6-7 µm (n.r. 2.0-3.0 µm)
                    (Figure 1); no leukocyte inclusions were observed. BC
                    revealed following data: platelet count 41x109/L [nor-        LOW C-MPL EXPRESSION AND IMPAIRED TPO
                    mal range (n.r.): 150-450], mean-platelet-volume              SIGNALING PATHWAYS IN ANKRD26-RELATED
                    (MPV) 13.6 fL (n.r.: 6.0-13.0). Hereditary macrothrom-        THROMBOCYTOPENIA: THERAPEUTIC
                    bocytopenia was, then, hypothesized. NGS demonstrat-          IMPLICATIONS.
                    ed the presence of heterozygosis in the MYH9 gene of
                    allele variant c.5629C>T, responsible for the novel           S. Barozzi1, F. Melazzini1,2, V. Bozzi1, F. Fumoso1,
                    p.Arg1877Trp amino acid exchange, supporting the              C. Zaninetti3, A. Pecci1,2
                    diagnosis of May-Hegglin disorder. The mature protein
                                                                                  1Medicina Generale, IRCCS Fondazione Policlinico San
                    shows a replacement of a very basic residue (arginine)
                    with a nonpolar residue (tryptophan), possibly impact-        Matteo, Pavia, Italy; 2Department of Internal Medicine,
                    ing myosin-9 structural features. Indeed, in silico anno-     University of Pavia, Italy; 3Institut für Immunologie und
                    tations indicated a p.Arg1877Trp deleterious effect on        Transfusionsmedizin, Greifswald, Germany
                    protein structure (Polyphen-2 score: 1.00; SIFT score:
                    0). Of note, according to the American College of             Background and Aims: ANKRD26-related thrombocy-
                    Medical genetics (ACMG) recommendations, this vari-           topenia (ANKRD26-RT) is an autosomal-dominant
                    ant is likely to be pathogenic, as it involves a mutational   thrombocytopenia associated with predisposition to




                                                                                                          ly
                    residue. The second case was a 57-years-old asympto-          myeloid malignancies. Patients present mild to severe
                    matic woman with two previous at term pregnancies in          thrombocytopenia with normal platelet size and function,




                                                                                                  on
                    absence of any bleeding complications. She was                mild bleeding tendency, and increased serum throm-
                    referred because of a failed response to TPO mimetics         bopoietin (TPO) concentrations. The disorder is caused
                    after an ITP diagnosis. The patient underwent a series of     by single nucleotide substitutions in the 5’UTR of the




                                                                                             e
                    diagnostic tests showing a platelet count of 20x109/L         ANKRD26 gene. A previous study showed that the vari-
                    and MPV value of 22.7 fL. Morphological examination
                    confirmed a larger platelet size [6-7 µm (n.r. 2-3 µm)].
                    However, clinical judgment guided toward the suspi-
                                                                                      us
                                                                                  ants interfere with the physiologic repression of
                                                                                  ANKRD26 during megakaryocyte differentiation, result-
                                                                                  ing in its persistent expression in late megakaryocytes:
                    cion of a congenital macrothrombocytopenia, mostly
                                                                                  al
                                                                                  this, in turn, associates with hyperactivation of the
                    because of the poor response to classical ITP therapies.      MAPK/ERK pathway and reduced proplatelet formation.
                                                                          ci

                    NGS investigation showed a heterozygous allele variant        We recently conducted a clinical trial on the efficacy of
                    within the ACTN1 gene, coding for actinin alpha 1. The        the TPO-mimetic Eltrombopag in inherited thrombocy-
                                                                    er


                    woman showed the allele variation c.1349G>A, caus-            topenias (ITs). The study showed a significant variability
                    ing p.Arg450His. The variation has been previously            in the response to the drug among ANKRD26-RT
                                                              m



                    described in subjects with a patient’s phenotype and to       patients; moreover, overall, ANKRD26-RT showed a
                    impair actinin alpha-1 dimerization and, in turn, platelet    lower extent of response compared to other forms of IT.
                                                  om




                    shape and internal organization. Conclusions:                 We therefore conducted a systematic analysis of the TPO
                    Massively sequencing technologies, such as the NGS, is        signaling pathways in the largest series of patients with
                    improving diagnostic approaches and enabling for              ANKRD26-RT analyzed for this aspect, including sub-
                    important clinical applications in the context of heredi-
                                               -c




                                                                                  jects who had received Eltrombopag within the clinical
                    tary thrombocytopenia, whose genetic determinants are         trial. Methods: We studied platelets from 11 ANKRD26-
                    rapidly expanding.
                                       on




                                                                                  RT patients, 7 of whom had participated in the trial. We
                                                                                  analyzed the phosphorylation state of the main kinases of
                                                                                  TPO signaling, STAT5, ERK and AKT, in basal condi-
                                  N




                                                                                  tions and after stimulation with TPO or TPO plus
                                                                                  Eltrombopag. Platelet expression of c-MPL, the receptor
                                                                                  for TPO, was measured by immunoblotting. Results: In
                                                                                  resting conditions, ANKRD26-RT platelets showed a sig-
                                                                                  nificantly higher activation of STAT5 and ERK com-
                                                                                  pared to control platelets (p=0.01). The analysis of the
                                                                                  same pathways following stimulation with TPO and
                                                                                  Eltrombopag showed a markedly reduced response to the
                                                                                  stimuli of patients’ platelets, expressed as the ratio
                                                                                  between induced and baseline phosphorylation. The acti-
                                                                                  vation of STAT5 in ANKRD26-RT platelets was reduced
                                                                                  by 80% following stimulation with TPO (p<0.005) and
                                                                                  84% after stimulation with TPO and Eltrombopag
                                                                                  (p<0.005) compared to control platelets. Similarly, ERK
                                                                                  activation was reduced by 51% (p<0.05) after TPO stim-
                                                                                  ulation and by 67% (p<0.005) when Eltrombopag was
                                                                                  added to TPO. We then explored whether these alter-
                    Figure 1.                                                     ations were associated with altered c-MPL expression.




                                                               [Hematology Reports 2021; 13:s3]                                            [page 111]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 112



         Poster




                    Indeed, c-MPL was found to be significantly reduced in         displayed defective delta granule content and were clas-
                    ANKRD26-RT platelets, with a mean decrease of 63%              sified as δ-storage pool disease (δ-SPD) (Figure 1). T-
                    (p<0.01) compared to controls. When we analyzed the            TAS compared favourably with lumi-aggregometry in
                    patients previously treated with Eltrombopag, we found         diagnosing the most severe forms of platelet function
                    a direct correlation between c-MPL expression and the          defects (i.e., δ-SPD) [test agreement (Lumi-LTA-vs-T-
                    extent of response to the drug in vivo (R2=0.73, p<0.01).      TAS) for the δ-SPD subgroup amounted to 80% and K
                    Conclusions: In ANKRD26-RT, the constitutive activa-           CHOEN to 0.695. T-TAS was less sensitive to the
                    tion of signaling downstream c-MPL is associated with a        milder defects [i.e., primary secretion defects (PSD)].
                    reduced expression of the receptor itself in platelets; this   Concerning patients on antiplatelet drugs, test agree-
                    results in a defective response to stimulation with c-MPL      ment (Lumi-aggregometry-vs-T-TAS) in detecting
                    ligands, such as TPO and Eltrombopag. Of note, we              patients, who were responders to the therapy amounted
                    found a direct relationship between the amount of c-MPL        to 54% and K CHOEN to 0.150. Conclusions: Results
                    expressed in platelets and the response to Eltrombopag in      indicate that T-TAS can diagnose the more severe forms
                    vivo, providing a plausible explanation for the variability    of platelet function defects (i.e., δ-SPD). There is limit-
                    of the response to the drug among ANKRD26-RT indi-             ed agreement T-TAS vs Lumi-aggregometry to identify
                    viduals. Finally, the reduced expression of c-MPL on           responders to antiplatelet drugs, but this poor agree-
                    platelet surface could result in a lower internalization of    ment is commonly shared by Lumi-aggregometry and
                    TPO, contributing to the markedly increased serum TPO          other devices owing to the lack of test specificity and
                    levels observed in ANKRD26-RT patients.                        prospective data from clinical trials linking platelet




                                                                                                           ly
                                                                                   function and therapeutic efficacy.




                                                                                                    on
                      PO048




                                                                                               e
                    FLOW-CHAMBER DEVICE, (T-TAS) TO DIAGNOSE
                    PATIENTS SUSPECTED OF HAVING PLATELET
                    FUNCTION DEFECTS                                                   us
                    S. La Marca1, A. Lecchi1, L. Padovan1, M. Boscarino1,
                                                                                   al
                    F. Peyvandi1,2, A. Tripodi1
                                                                           ci

                    1Fondazione   IRCCS Ca’ Granda Ospedale Maggiore
                                                                    er


                    Policlinico, Angelo Bianchi Bonomi Hemophilia and
                    Thrombosis Center and Fondazione Luigi Villa, Milano,
                                                               m



                    Italy; 2Università degli Studi di Milano, Department of
                    Pathophysiology and Transplantation, Milano, Italy
                                                  om




                    Background and Aims: Patients suspected of having
                    platelet defects represent a diagnostic challenge for the
                    clinical laboratory, mainly due to the complexity and
                                               -c




                    poor standardization of methods used for investigation.
                                       on




                    We provide herewith a comparative face-to-face results
                    evaluation for a new flow-based chip-equipped point-
                    of-care device, Total-Thrombus-Analysis-System (T-
                                  N




                    TAS), meant to measure the overall primary hemostatic
                    function vs the more complicated and time consuming
                    lumi-aggregometry and platelet delta granule measure-
                    ment. Patient population: Object of the study was a
                    group of patients (n=96) suspected of having platelet
                    defects. They displayed a mild-to-moderate bleeding            Figure 1.
                    diathesis of variable severity, characterized by mucocu-
                    taneous bleeding, menorrhagia, and excessive post-sur-
                    gical and post-traumatic blood loss. This cohort was           PO049
                    characterized as having the following parameters within
                    normal limits: platelet count and size; prothrombin and
                    activated partial thromboplastin time; VWF (antigen            PLATELET DISTRIBUTION WIDTH AS A
                    and ristocetin cofactor activity) and factor VIII activity.    MORTALITY RISK PREDICTOR: PROSPECTIVE
                    We also investigated a second group of patients (n=26)         FINDINGS IN AN ITALIAN ADULT GENERAL
                    referred for the evaluation of residual platelet hyperac-      POPULATION
                    tivity whilst on antiplatelet therapy. Results: Forty-
                    eight (PFDs) of 96 patients (BP) displayed abnormal            S. Costanzo1, B. Izzi1, A. Gialluisi1,2, A. De Curtis1,
                    platelet function by lumi-aggregometry and 10 of the 48        S. Magnacca3, T. Panzera1, A. Di Castelnuovo3,




         [page 112]                                        [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 113



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    F. Gianfagna2,3, M.B. Donati1, C. Cerletti1,
                    G. de Gaetano1, L. Iacoviello1,2,
                    for the Moli-sani Study Investigators
                    1Department   of Epidemiology and Prevention, IRCCS
                    NEUROMED, Pozzilli, IS, Italy; 2EPIMED Research
                    Center, Department of Medicine and Surgery, University
                    of Insubria, Varese, Italy; 3Mediterranea Cardiocentro,
                    Napoli, Italy

                    Background and Aims: Platelet Distribution Width
                    (PDW) is a marker of platelet size heterogeneity recog-      Figure 1.
                    nized as a diagnostic marker for diseases with impaired
                    megakaryocyte differentiation and thrombopoiesis, as it
                    represents a readout from the combination of processes       PO050
                    leading to platelet production and destruction. Recently,
                    PDW was also found to tag platelet activation variability,
                    both ex vivo and in vitro and to be a prognosis predictor    IMPORTANCE OF THE SCREENING FOR
                    of cardiovascular disease (CVD), cancer, diabetes and        VARIANTS CAUSING INHERITED




                                                                                                         ly
                    neurological disorders. We therefore evaluated the possi-    THROMBOCYTOPENIAS PREDISPOSING TO
                    ble relationship, at population level, between PDW val-      HEMATOLOGICAL MALIGNANCIES IN POTENTIAL




                                                                                                  on
                    ues and all-cause and cause-specific mortality. Methods:     BONE MARROW DONORS, A CASE REPORT
                    Longitudinal analysis was performed on 17,334 partici-
                    pants (52% women, mean age 55.6 ±12 years) from the          L. Bury1, E. Falcinelli1, A.M. Mezzasoma1, A. Carotti2,




                                                                                             e
                    Moli-sani study (enrollment 2005-2010), without a histo-     P. Gresele1
                    ry of haematological diseases and for whom baseline
                    PDW measurements were available. All-cause and
                    cause-specific mortality was validated by using the
                                                                                     us
                                                                                 1Department  of Medicine and Surgery, Section of
                                                                                 Internal and Cardiovascular Medicine, University of
                                                                                 al
                    Italian Death Registry. PDW measurements were catego-        Perugia, Perugia, Italy; 2Department of Medicine and
                    rized in tertiles, the first acting as the reference.        Surgery, Division of Hematology and Clinical
                                                                         ci

                    Multivariable Cox-proportional hazard model was used         Immunology, University of Perugia, Perugia, Italy
                    to estimate the association of PDW and mortality, using
                                                                   er


                    multiple imputation technique for missing data. Results:     Background and Aims: Inherited platelet disorders
                    In the whole population sample, the mean value of PDW        predisposing to hematological malignancies are autoso-
                                                             m



                    was 16.4 ±0.58 fL. Over a median follow-up of 11.6           mal dominant forms characterized by thrombocytope-
                    years (interquartile range 10.7-12.5), 1,535 deaths          nia, most often mild, with normal or slightly increased
                                                 om




                    (37.7% CVD and 36.5% cancer) were identified. As             platelet volume, a bleeding diathesis of widely variable
                    compared to those in the first tertile of PDW (14.6-16.0     severity and an enhanced risk of developing hematolog-
                    fL), individuals within the highest tertile (16.6-20.4 fL)   ical malignancies. Three disorders have been identified
                                              -c




                    had an increased risk of all-cause (HR: 1.20; 95% CI:        so far: familial platelet disorder with propensity to acute
                    1.04-1.37) and CVD mortality (HR: 1.29; 95% CI: 1.03-        myelogenous leukemia (FPD-AML), ETV6-related
                                       on




                    1.62). (Figure 1). In contrast, no association between       thrombocytopenia (ETV6-RT) and ANKRD26-RT.
                    PDW and cancer mortality was found. Additionally, all-       FPD-AML is caused by mutations of RUNX1, a tran-
                    cause and CVD mortality increased by 5% and 7%,              scription factor crucial for megakaryocyte maturation.
                                  N




                    respectively, for each increase in 1 standard deviation      About 45% of FPD-AML patients evolve to a hemato-
                    (0.58 fL) of PDW (HR: 1.05; 95% CI:1.00-1.10 and HR:         logical malignancy during their life, mostly to myeloid
                    1.07; 95% CI: 0.99-1.16, respectively). No difference in     leukemia or myelodysplastic syndromes (MDS), but
                    the association between PDW and all-cause and CVD            also to T-cell or B-cell lymphoproliferative disorders.
                    mortality was observed by subgroup analyses by gender,       Next-generation sequencing (NGS) has revolutionized
                    history of CVD/cancer and major risk factors of non-         the diagnostic approach to these disorders providing a
                    communicable diseases. However, subgroup analyses by         rapid molecular diagnosis but it has also raised discus-
                    age classes (35-65 years, ≥65 years) showed that the         sion about the ethical implications of discovering unex-
                    association of PDW with all-cause mortality was more         pected variants associated with leukemic risk. We
                    evident in the elderly (HR: 1.34; 95% CI: 1.14-1.58, p for   report a case showing the importance of the prompt
                    interaction=0.028, Figure 1). Similar results were found     identification of these variants. Methods: Platelet func-
                    when cancer mortality was investigated (age ≥65 years:       tion was studied by light transmission aggregometry
                    HR: 1.37; 95% CI: 1.01-1.85, p for interaction=0.020).       (LTA), flow cytometry and lumiaggregometry. Variants
                    Conclusions: A PDW-associated increase of all-cause          affecting genes involved in bleeding and platelet disor-
                    and CVD mortality risk, could be related to                  ders were assessed by NGS, performing targeted
                    accelerated/altered platelet activation, production or       sequencing of 91 TIER-1 genes. Germinal variants
                    destruction, all age-dependent processes leading to sev-     were confirmed by Sanger sequencing of DNA extract-
                    eral clinical conditions and death.                          ed from finger nails. Results: A patient came to our




                                                              [Hematology Reports 2021; 13:s3]                                             [page 113]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 114



         Poster




                    attention for mild thrombocytopenia found on the occa-       recommended for patients with polycythemia vera
                    sion of a blood donation. He did not refer hemorrhagic       (PV), a myeloproliferative neoplasm (MPN) with
                    manifestations. We confirmed very mild thrombocy-            increased risk of arterial and venous thromboses. Based
                    topenia (121x109/L) associated with defective platelet       on aspirin pharmacodynamics in essential thrombo-
                    aggregation in response to adrenaline, U46619 and col-       cythemia, a twice-daily regimen is recommended for
                    lagen and defective content and release of α and δ gran-     PV patients deemed at particularly high thrombotic risk.
                    ules. Expression of platelet glycoproteins was normal.       Methods: We investigated the effects of low-dose
                    NGS identified a rare variant in RUNX1, p.Gln262Ter,         aspirin on platelet cyclooxygenase activity and in vivo
                    classified as pathogenic in ClinVar and present in the       platelet activation in 49 PV patients, as assessed by
                    literature and was then diagnosed FPD-AML. Our               serum thromboxane (TX)B2 and urinary TXA2/TXB2
                    patient reported that his brother had been affected by       metabolite (TXM) measurements, respectively. A previ-
                    AML in the ‘90s and was treated by allogeneic                ously described pharmacokinetic-pharmacodynamic in
                    hematopoietic stem cells transplantation for which he        silico model was used to simulate the degree of platelet
                    was the donor. We therefore wondered if the brother          TXA2 inhibition by once-daily (qd) and twice-daily
                    with AML who received his bone marrow was also a             (bid) aspirin, and to predict the effect of missing an
                    carrier of the same genetic variant. Given that the broth-   aspirin dose during qd and bid regimens. Results:
                    er circulating white blood cells would necessarily pres-     Serum TXB2 averaged 8.2 [1.6-54.7] ng/ml and signifi-
                    ent the mutation, we analyzed DNA from his finger            cantly correlated with the platelet count (rho=0.39) and
                    nails and found the same RUNX1 variant. Platelet func-       urinary TXM (rho=0.52) in multivariable analysis.




                                                                                                         ly
                    tion tests revealed the same type of platelet function       One-third of aspirin-treated PV patients displayed less-




                                                                                                  on
                    defect observed in the bone marrow donor brother.            than-maximal platelet TXB2 inhibition, and were char-
                    Conclusions: The study of this family shows that the         acterized by significantly higher platelet counts and
                    brother of our propositus developed AML because car-         platelet-count corrected serum TXB2 than those with
                    rier of FPD-AML and he received hematopoietic stem           adequate inhibition. Eight PV patients were sampled




                                                                                            e
                    cell transplantation from our propositus carrier of FPD-     again after 12±4 months, and had reproducible serum
                    AML too. This poses the patient at risk of further
                    leukemic transformation. At the time of transplantation
                    (1990s) FPD-AML was still unknown but nowadays we
                                                                                     us
                                                                                 TXB2 and urinary TXM values. The in silico model pre-
                                                                                 dicted complete inhibition of platelet-derived TXB2 by
                                                                                 bid aspirin, a prediction verified in a PV patient with the
                                                                                 al
                    know that platelet disorders predisposing to hematolog-      highest TXB2 value while on aspirin qd and treated
                    ical malignancy are more common than expected,               short-term with a bid regimen. Conclusions: In conclu-
                                                                            ci

                    therefore: 1) after obtaining explicit informed consent,     sion, one in three PV patients on low-dose aspirin dis-
                    patients with suspected platelet disorders predisposing      play less-than-maximal inhibition of platelet TXA2 pro-
                                                                   er


                    to hematological malignancies should be genetically          duction. A personalized approach to antiplatelet therapy
                    screened for these disorders; 2)mutational screening of      can be guided by serum TXB2 measurements.
                                                             m



                    siblings should be promptly performed to avoid to
                    transplant patients with cells carrying the same genetic
                                                 om




                    variant.                                                     PO052

                                                                                 EFFICACY OF THROMBOPOIETIN RECEPTOR
                                              -c




                      PO051
                                                                                 AGONISTS IN PATIENTS WITH IMMUNE
                                                                                 THROMBOCYTOPENIA AND MEGAKARYOCYTES
                                       on




                    PLATELET THROMBOXANE INHIBITION BY                           WITH MYELODYSPLASTIC FEATURES
                    LOW-DOSE ASPIRIN IN POLYCYTHEMIA VERA:
                                  N




                    EX VIVO AND IN VIVO MEASUREMENTS AND IN                      R. Tomasello1, F. Romano1, M. Mattana1, C. Rotolo1,
                    SILICO SIMULATION                                            C. Cammarata1, A. Lucchesi2, G. Giordano3,
                                                                                 S. Siragusa1, M. Napolitano1
                    G. Petrucci1, A. Giaretta2, P. Ranalli3, V. Cavalca4,
                    A. Dragani3, B. Porro4, D. Hatem1, A. Habib5,                1Department  of Health Promotion, Mother and Child
                    E. Tremoli6, C. Patrono1, B. Rocca1                          Care, Internal Medicine and Medical Specialties
                                                                                 (ProMISE), University of Palermo, Palermo, Italy;
                    1Department    of Safety and Bioethics, Section of           2IRCCS Istituto Romagnolo per lo studio dei tumori

                    Pharmacology, Catholic University School of Medicine,        (IRST)”Dino Amadori”, Meldola (FC), Italy; 3Division
                    Rome, Italy; 2Department of Pathology, University of         of Internal Medicine, Hematology Service, Regional
                    Cambridge, Cambridge, UK; 3Department of                     Hospital “ACardarelli”, Campobasso, Italy
                    Hematology, SSpirito Hospital, Pescara, Italy; 4Centro
                    Cardiologico Monzino, IRCCS, Milano, Italy;                  Background and Aims: Immune Thrombocytopenia
                    5Department of Basic Medical Sciences, QU Health,            (ITP) is a bleeding disorder characterized by persist-
                    Qatar University, Doha, Qatar; 6Maria Cecilia Hospital,      ently low platelet counts (PLT). ITP is a diagnosis of
                    Cotignola, Italy                                             exclusion that may require bone marrow (BM) exami-
                                                                                 nation to rule out a unilinear myelodysplastic syn-
                    Background and Aims: Low-dose aspirin is currently           drome (MDS). BM histopathology of patients with ITP




         [page 114]                                       [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 115



                                                                       XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    usually shows megakaryocyte (MK) hyperplasia with          Table 1.
                    aggregation of cells to form diffuse clusters, sometimes
                    associated to mature lymphoid cells infiltration. In a
                    few cases, BM examination shows signs of MK dys-
                    plasia with no other criteria compatible with MDS.
                    There is currently no difference between the treatment
                    of patients with BM compatible with ITP and those
                    with isolated myelodysplastic features of MKs.
                    Methods: We performed a retrospective analysis that
                    included all patients diagnosed with ITP from 2011 to
                    2021, treated with thrombopoietin receptor agonists
                    (TPO-RAs) as 2nd (or further) line treatment, whose
                    BM examination (performed before TPO-RA adminis-
                    tration) showed signs of MK dysplasia. Treatment with      PO053
                    TPO-RAs started in this group of patients in a period
                    ranging from 2016 to 2021. Patients with a prior diag-     PLATELET FUNCTION IN PATIENTS WITH
                    nosis of MDS, HCV-related liver disease and autoim-        PTEN-MUTATIONS
                    mune diseases were excluded from the study. All
                    patients included that were not treated with TPO-RAs       S. Sorrentino1, A. Pasquarella2, R. Onesimo2, C. Leone2,




                                                                                                        ly
                    on first relapse ultimately developed resistance to        A. Curatola2, G. Zampino2,3, I. Lazzareschi2,3,
                    steroids (PLT persistently <50000/mmc under treat-         E. De Candia1,4




                                                                                                on
                    ment with maximum tolerable dose of steroid) or intol-
                    erance (to the minimum effective dose of steroid able      1Unità Malattie Emorragiche e Trombotiche, Fondazione
                    to maintain a PLT >50000/mmc), some of them were           Policlinico Universitario Agostino Gemelli IRCSS, Roma,




                                                                                          e
                    also unresponsive to Rituximab and splenectomy. We         Italy; 2UOC Pediatria, Fondazione Policlinico
                    evaluated response to TPO-RAs at 2 weeks, 3 months,
                    6 months and 1 year from start of treatment. Results:
                    We evaluated 16 patients of age at diagnosis ranging
                                                                                   us
                                                                               Universitario Agostino Gemelli IRCSS, Roma, Italy;
                                                                               3Dipartimento di scienze della Vita e della Sanità Pubblica,

                                                                               Università Cattolica del Sacro Cuore, Roma, Italy;
                                                                               al
                    from 40 to 83yo (median age 58.5y), M:F ratio of           4Dipartimento di Medicina e Chirurgia Traslazionale,
                    1.3:1, diagnosed with ITP who became unresponsive to       Università Cattolica del Sacro Cuore, Roma, Italy
                                                                       ci

                    steroids and were ultimately treated with either
                    Eltrombopag (ETP, n=8) or Romiplostim (RPS, n=8).          Background and Aims: Phosphatase and tensin
                                                                 er


                    In all patients, a BM examination showed myelodys-         homolog (PTEN) Hemartoma Tumor Syndrome (PHTS)
                    plastic features of MKs. All patients were treated with    is a group of very rare diseases due to mutations in PTEN
                                                            m



                    corticosteroids with or without high dose intravenous      gene and characterized by amartomatosis tumors. A pre-
                    immunoglobulins (IVIGs) on first line. Three patients      vious study in mouse models showed that PTEN is
                                                om




                    started TPO-RAs in 2nd line. Twelve patients started       involved in the negative regulation of collagen-induced
                    TPO-RAs in 3rd or further lines due to a loss of           platelet activation As a consequence, PTEN-deficient
                    response to previous immunosuppressive treatments.         mice had increased platelet aggregation to collagen com-
                                             -c




                    Two patients treated with ETP experienced sustained        pared to control mice (Weng, 2010). Due to a collabora-
                    response (PLT >100000/mmc for more than 3 months           tion with the Rare Diseases Center in our institution, we
                                      on




                    off therapy). Response at 2 weeks and 1 year were bet-     could perform a pilot study on PHTS patients with the
                    ter in the ETP group. BM aspiration and biopsy were        aim of evaluating platelet function in these patients, with
                    performed again on 2 patients out of 15 due to devel-      a special focus on platelet aggregation by low dose col-
                                 N




                    opment of other cytopenias, after 2 years and 1 year of    lagen. Methods: The study population included 12
                    treatment with TPO-RAs respectively, and showed            patients (with proven genetic diagnosis of PHTS and not
                    overt MDS (Table 1). Conclusions: Limited to the           in treatment with antiaggregant/anticoagulant drugs) and
                    small number of patients enrolled, this study demon-       11 controls (volunteer patients with no PHTS diagnosis).
                    strates the efficacy of long-term treatment with TPO-      Platelet function was investigated using: 1) PFA 100
                    RAs in patients with underlying MK myelodysplastic         System collagen/epinephrine (CEPI) and collagen/ADP
                    features in the BM observed at diagnosis. The overall      (CADP) test cartridges, 2) light transmission aggregom-
                    response to immunosuppressive treatments such as           etry (LTA) and platelet rich plsma (PRP) stimulated by
                    Rituximab or steroids appears to be short lived and        ADP (2 µM), collagen (2, 1, 0.5 µg/ml), epinephrine (5,
                    hard to maintain in the analyzed cohort, thus suggest-     2.5, 1.25, 0.6 µM). Maximal aggregation (MA, %) for all
                    ing a role of MK dysplasia in the mechanism of throm-      agonists and lag phase (LP, seconds) for collagen were
                    bocytopenia in addition to autoimmunity. Due to their      measured (Figure 1). Results: Platelet aggregation by
                    mechanism of action, TPO-RAs actually represent the        ADP 2 µM was not significantly different in PTEN
                    optimal treatment for these patients, as they can stimu-   patients compared to the control group (MA >75%, data
                    late thrombopoiesis while also inducing a degree of        not shown). Platelet aggregation by collagen 2 µg/ml, 1
                    immunotolerance. Furthermore, this study shows the         µg/ml and 0.5 µg/ml was >70% in both populations with
                    possibility of a sustained response in patients affected   a longer LP in PTEN patients (LP >70 sec), however no
                    by ITP with signs of MK dysplasia.                         statistically significant differences were found. Platelet




                                                             [Hematology Reports 2021; 13:s3]                                             [page 115]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 116



         Poster




                    aggregation by low dose epinephrine, i.e. 5, 2.5, 1.25 and   PO054
                    0.6 µM also was not different in PTEN patients com-
                    pared to controls. PFA-100 CEPI and CADP closure
                    times were prolonged in 67% and 50% of PTEN patients,        IDENTIFICATION OF A TUBB1 MISSENSE
                    respectively (normal values CEPI <140 seconds; CADP          MUTATION IN A CONGENITAL
                    <120 seconds). Conclusions: Although previous studies        MACROTHROMBOCYTOPENIA FAMILY BY
                    in animal models suggested that PTEN regulation of           CLINICAL EXOME SEQUENCING (CES)
                    PI3K/Akt pathway could negatively affect collagen-
                    induced platelet aggregation, we did not confirm             G. D’Andrea, A. Ariano, M. D’Apolito, G. Cordisco,
                    increased aggregation by low collagen concentrations in      I. Adipietro, G.P. Palladino, M.F. D’Ambrosio,
                    patients with PTEN-related diseases. Despite the small       M.A. Di Stefano, M. Curcetti, R. Santacroce,
                    number of patients analyzed, this pilot study is the first   M. Margaglione
                    study in human subjects affectd by these very rare dis-
                    eases. Our results suggest that PTEN could regulate col-     Genetica Medica, Dipartimento di Scienze Mediche e
                    lagen-induced aggregation to a lesser extent in humans       Sperimentali, Università di Foggia, Foggia, Italy
                    rather than in mice. Studies on a larger number of
                    patients and using a wider range of agonists concentra-      Background: Congenital macrothrombocytopenia is a
                    tions are needed to elucidate the role of PTEN in the reg-   heterogeneous group of rare disorders with an autoso-
                    ulation of platelet function.                                mal-dominant inheritance, characterized by increased




                                                                                                         ly
                                                                                 platelet size and a decreased number of circulating
                    Reference:                                                   platelets.Several genes are associated with macrothrom-




                                                                                                 on
                    Weng Z, Li D, Zhang L, Chen J, Ruan C, Chen G,               bocytopenia, in particulary, the genes implicated, encode
                       Gartner TK, Liu J. PTEN regulates collagen-               proteins that vary widely in function and include tran-
                       induced platelet activation. Blood. 2010 Oct              scription factors, proteins involved in cytoskeleton




                                                                                            e
                       7;116(14):2579-81. doi: 10.1182/blood-2010-03-            rearrangement and organization.One of less frequent
                       277236.
                                                                                     us
                                                                                 forms of macrothombocytopenia is determined by com-
                                                                                 ponent defects of the cytoskeleton that regulate pro-
                                                                                 platelet formation and morphology through microtubules
                                                                                 al
                                                                                 compounds of α/β-tubulin. Abnormal assembly and
                                                                                 incorporation of β1-Tubulin is caused by molecular
                                                                         ci

                                                                                 defects of TUBB1 gene.The aims of this study were to
                                                                                 characterize molecularly a family affected by
                                                                   er


                                                                                 macrothrombocytemia using a new sequencing method
                                                                                 (CES), then, to validate its importance in the molecular
                                                             m



                                                                                 diagnostics of genetically heterogeneous pathology.
                                                                                 Materials and Methods: This report describes a 15
                                                 om




                                                                                 year-old girl with macrothrombocytopenia, diagnosed
                                                                                 from the first years of the life and inherited from mother
                                                                                 family branch. Indeed,The mother and a maternal aunt
                                              -c




                                                                                 are platelets deficiency too,The young men, after
                                                                                 obtainig the parental consent informed, was subjected to
                                       on




                                                                                 a molecular investigation. DNA was extracted from
                                                                                 peripheral blood leukocytes and subsequently submitted
                                                                                 to an Clinical Exome Sequencing (CES) analysis by a kit
                                  N




                                                                                 DNAPrep with enrichment Illumina on platform
                                                                                 NextSeq. 550.The results, at first, were analyzed by an
                                                                                 Illumina Variant Interpret platform,then,to verify candi-
                                                                                 date mutations and examine their segregation among
                                                                                 family members, Sanger sequencing was performed
                                                                                 using standard methods on an ABI 3130 automated
                                                                                 sequence. Results: Molecular analysis revealed a het-
                                                                                 erozygous variant p.Gln191Pro in the exon 4 of TUBB1
                                                                                 gene with unknown clinical significance (VUS). Direct
                                                                                 Sanger sequencing confirmed the mutation both in the
                                                                                 patient and in the mother, but none in the maternal aunt
                                                                                 due to the DNA unavailability. In silico analysis as
                                                                                 Polyphen-2 and SIFT Web Server assign to the mutation
                                                                                 an pathogenic effect on β1-tubulin protein.The replace-
                                                                                 ment of a polar amino acid with a non-polar one consti-
                                                                                 tutes a destabilizing element of the tertiary structure of
                                                                                 protein and determines, probably, deficient functional
                    Figure 1.                                                    microtubules that might lead to defective proplatelet for-




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 117



                                                                            XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    mation and abnormal protrusion-like platelet release,            nia. We believe that this drug can serve as a potential
                    resulting in macrothrombocytopenia. Conclusions: Our             treatment option when a short period of platelet increase
                    results underline the usefulness of the Clinical Exome           is needed for clinical reasons other than invasive proce-
                    Sequencing in clinical practice as well as that it is an effi-   dures in patients with chronic liver disease complicated
                    cient method for a quick and a cheaper molecular analy-          by thrombocytopenia.
                    sis of pathogenic variants in inherited macrothombocy-
                    topenia or for most heterogeneous genetically patholo-
                    gies. In any case, family segregation and in vitro molec-        PO056
                    ular studies are indispensable to demonstrate the effec-
                    tive role of the mutation in the development of
                                                                                     CORRELATION BETWEEN BLEEDING SEVERITY
                    macrothrombocytopenia in the family we have analyzed.
                                                                                     AND LABORATORY TEST RESULTS IN PATIENTS
                                                                                     WITH INHERITED PLATELET FUNCTION
                    PO055                                                            DISORDERS (THE BAT-LAB SUBSTUDY)

                                                                                     E. Falcinelli1, M.C. Alessi2, L. Bury1, G. Podda3,
                    LUSUTROMBOPAG TREATMENT IN A PATIENT                             G. Guglielmini1, M. Fiore4, T. Sevivas5, N. Bermejo6,
                    WITH THROMBOCYTOPENIA CIRRHOSIS                                  E. De Candia7,8, M. Chitlur9, M. Lambert10,
                    RELATED PRIOR TO RADIOTHERAPY FOR                                L. Barcella11, P.G. Heller12,13, C. Falaise2, P. Gresele1




                                                                                                             ly
                    OROPHARYNGEAL SQUAMOUS CELL
                    CARCINOMA: A CASE REPORT                                         1Department of Medicine and Surgery, Section of Internal




                                                                                                      on
                                                                                     and Cardiovascular Medicine, University of Perugia,
                    F. Valeri1, C. Dainese1, M. Bardetta1, C. Sella1,                Italy; 2Centre for CardioVascular and Nutrition research
                    B. Bruno2, A. Borchiellini1                                      (C2VN), INSERM 1263, INRA 1260, Marseille, France;




                                                                                                e
                                                                                     3Medicina III, ASST Santi Paolo e Carlo, Dipartimento di
                    1CRR  Malattie Emorragiche e Trombotiche dell’Adulto,
                    Ematologia Universitaria, AOU Città della Salute e
                    della Scienza di Torino; 2Ematologia Universitaria,
                                                                                         us
                                                                                     Scienze della Salute, Università degli Studi di Milano;
                                                                                     4Laboratory of Hematology, University Hospital of

                                                                                     Bordeaux, Pessac, France; 5Servicio de Hematologia
                    Dipartimento di Oncologia, AOU Città della Salute e
                                                                                     al
                                                                                     Clinica- Centro Hospitalar e Universitario de Coimbra,
                    della Scienza di Torino, Italy                                   Coimbra, Portugal; 6Hospital San Pedro de Alcantara,
                                                                             ci

                                                                                     Caceres, Spain; 7Hemostasis and Thrombosis Unit,
                    Background: Lusutrombopag (Mulpleo®) is an orally                Fondazione Policlinico Agostino Gemelli IRCCS, Roma;
                                                                      er


                    bioavailable, small molecule thrombopoietin receptor             8Institute of Internal Medicine and Geriatrics, Università
                    agonist approved for the treatment of thrombocytopenia           Cattolica del Sacro Cuore, Roma; 9Children’s Hospital of
                                                                m



                    in adult patients with chronic liver disease who are             Michigan, Detroit, USA; 10Children’s Hospital of
                    scheduled to undergo a procedure. In placebo-controlled          Philadelphia, Philadelphia, USA; 11Immunohematology
                                                   om




                    phase 3 clinical trials, lusutrombopag (at the dose of 3-        and Transfusion Medicine (SIMT) & Hemostasis and
                    mg tablet once daily for up to 7 days) significantly             Thrombosis Center ASST Papa Giovanni XXIII, Bergamo,
                    increased the proportion of patients who did not require         Italy; 12Instituto de Investigaciones Médicas ALanari,
                                                -c




                    a platelet transfusion prior to the procedure or rescue          Facultad de Medicina, Universidad de Buenos Aires,
                    therapy for bleeding up to 7 days after the scheduled pro-       Buenos Aires, Argentina; 13Departamento Hematología
                                        on




                    cedure. In the present report, we discuss a case in which        Investigación, Consejo Nacional de Investigaciones
                    radiotherapy (who we considered comparable to proce-             Científicas y Tecnológicas (CONICET), Universidad de
                    dure) was performed reaching target platelet levels fol-         Buenos Aires, Instituto de Investigaciones Médicas
                                   N




                    lowing oral administration of lusutrombopag. Case                (IDIM), Buenos Aires, Argentina
                    Report: We report the case of a 69-year-old woman
                    affected by chronic liver disease HBV related who devel-         Background and Aims: The diagnostic approach to
                    oped an oropharyngeal squamous cell carcinoma. The               inherited platelet function disorders (IPFD) is based on
                    patient underwent first line chemotherapy (cisplatin             the bleeding history (ISTH-BAT) and on a streamlined
                    (CDDP) and 5-fluorouracil (5-FU) with a 30% reduction            panel of laboratory tests. Few studies have assessed the
                    dose due to a thrombocytopenia cirrosis related (basal           predictive value of platelet laboratory tests for bleeding
                    value 76 x 109/L) but this treatement was early stopped          manifestations in patients with IPFD, with inconclusive
                    because of worsening of thrombocytopenia (39 x 109/L).           results. This is an ad hoc designed substudy of the mul-
                    Oncologist decided then to shift treatement to radiother-        ticentric international ISTH BAT-validation study pro-
                    apy (RT), but platelet values were inadequate. 5 days            moted by the Platelet Physiology SSC of the ISTH
                    prior to radiotherapy, oral lusutrombopag was initiated at       (BAT-VAL) including patients with a well-defined diag-
                    a dosage of 3 mg/day (day 1) and was continued for 7             nosis of IPFD. Aim of the present study was to assess
                    days. Platelet count on day 5 (first day of RT) was 62 ×         whether there is an association between the bleeding
                    109/L and on day 21 (the last day of RT) was 70 × 109/L,         score (BS) and results of platelet function tests in
                    allowing the entire treatment. Conclusions: Use of               patients with IPFD. Methods: Participating centers were
                    lusutrombopag in this setting allowed our patient to safe-       asked to fill in a standardized form on laboratory results
                    ly perform radiotherapy despite initial thrombocytope-           of the platelet function studies performed to diagnose




                                                                 [Hematology Reports 2021; 13:s3]                                                [page 117]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 118



         Poster




                    the enrolled IPFD patients. Laboratory data included          1Magnetic  Resonance Center (CERM) and Department
                    bleeding time (BT), PFA-100® closure time, light trans-       of Chemistry “Ugo Schiff”, University of Florence, Sesto
                    mission aggregometry (LTA), alpha granule release and         Fiorentino, Italy; 2Consorzio Interuniversitario
                    platelet glycoprotein expression by flow cytometry.           Risonanze Magnetiche MetalloProteine (CIRMMP),
                    Results were classified as abnormal when out of the           Sesto Fiorentino, Italy; 3Department of Experimental
                    internal laboratory reference range. Results: 72 patients     and Clinical Medicine, University of Florence, Florence,
                    (37% of the initially enrolled IPFD population), with 16      Italy; 4IRCCS Fondazione Don Carlo Gnocchi,
                    different IPFD forms, were included. Median baseline          Florence, Italy; 5NEUROFARBA Department,
                    BS was 7.5 (IQR 2-12) (BS of healthy controls: 0, IQR         University of Florence, Florence, Italy
                    0-1). A LTA defect in response to at least one platelet
                    agonist was found in 71% individuals. The % of patients       Background and Aims: Atrial fibrillation (AF) is the
                    with high BS and defective LTA in response to ADP was         most common cardiac arrhythmia with a clinical rele-
                    significantly higher compared to that of patients with        vance observed in medical practice1. Long-term oral
                    normal BS and defective LTA (59.4±4.7% vs                     anticoagulation is the mainstay therapy for ischemic
                    35.5±4.8%, p<0.05). BT and PFA-100® closure time              stroke prevention in AF patients2. However, benefits of
                    were altered, respectively, in 64% and 88% of patients.       anticoagulation need to be carefully balanced against the
                    BT was significantly more prolonged in patients with a        risk of bleeding, with intracranial hemorrhage being the
                    high BS compared to those with a normal BS                    most dreaded scenario. Stroke risk in AF is not a homo-
                    (27.1±6.2min vs 15.1±10.6min, p<0.01), while PFA-             geneous phenomenon, currently decision making regard-




                                                                                                           ly
                    100® closure times was not different between patients         ing antithrombotic drug administration relays on avail-
                    with normal or high BS. A ROC curve analysis revealed         able stroke and bleeding risk stratification schemes (e.g.,




                                                                                                   on
                    moderate discriminative ability of the BT for pathologic      CHADS2VASC2, HAS-BLED scores) which are based
                    BS (AUC 0.795, p<0.001). Flow cytometry showed that           on clinical information3. However, their validity is still
                    27% of patients had abnormal αIIbβ3 expression and            controversial, thus the identification of novel stratifica-




                                                                                              e
                    39% defective αIIbβ3 activation (measured as PAC-1            tion approaches able to recognize patients at high risk of
                    binding). The % of patients with high BS and defective        bleeding is pivotal for AF patient management.
                    αIIbβ3 expression was significantly higher compared to
                    that of patients with normal BS and defective αIIbβ3
                                                                                      us
                                                                                  Methods: The Strat-AF study is a single center, longitu-
                                                                                  dinal observation study evaluating elderly AF patients on
                    (90% vs 10%, p<0.05). Moreover, the % of patients with        oral anticoagulation for thromboembolism prevention.
                                                                                  al
                    defective PAC-1 binding and pathologic BS was signif-         The goal of this project is to evaluate if biological mark-
                    icantly higher than that of patients with defective PAC-      ers, either clinical, circulating and neuroimaging-based,
                                                                          ci

                    1 binding but normal BS (66% vs 33%, p<0.05). A ROC           and their possible combinations, can improve the predic-
                                                                    er


                    curve analysis revealed a moderate discriminative abili-      tion of bleeding risk in AF patients under treatment with
                    ty of abnormal PAC-1 binding for pathologic BS (AUC           any type of oral anticoagulants4. Serum samples of 177
                    0.778, p<0.001). α-granules content was reduced in 29%        AF patients (mean age 78.2 years, 55.3% males, 70% on
                                                              m




                    of patients and was significantly more common among           DOAC treatment) were analyzed via high resolution 1H-
                    patients with a pathologic BS compared to those with a        NMR spectroscopy5. Data on 27 metabolites and 112
                                                  om




                    normal BS (80% vs 20%, p<0.05). Conclusions: This             lipoprotein-related fractions were obtained and analyzed
                    study shows that an abnormal ISTH-BAT BS is associ-           with multivariate and univariate statistical approaches.
                    ated with altered laboratory platelet function test results   Results: We characterized, for the first time to best of our
                                               -c




                    in IPFD. In particular, impairment of LTA in response to      knowledge, how different kinds of oral anticoagulation,
                    ADP, prolongation of the BT, reduced α-granules con-          Vitamin K and coumarin anticoagulants, affect the serum
                                       on




                    tent, reduced αIIbβ3 expression and decreased ADP-            metabolomic profiles of patients, obtaining a good dis-
                    induced PAC-1 binding were associated with increased          crimination between the two treatments (71% accuracy).
                    BS. Despite the relatively small patient number of this       Moreover, several statistically significant associations
                                  N




                    study suggests a predictive value of some platelet assays     (FDR        adjusted       p-value     <0.05)      between
                    for bleeding in patients with IPFD. An ad hoc designed        metabolomic/lipoproteomic features and clinical out-
                    prospective study on the predictive value of platelet         comes (i.e. memory disturbances, stroke events,
                    function tests for subsequent bleeding in patients with       CHADS2VASC2 score) emerged from the regression
                    IPFD is highly warranted.                                     analyses. Conclusions: Our results support the useful-
                                                                                  ness of NMR-based metabolomic approach in the AF set-
                                                                                  ting both for the description of the pharmacological
                      PO057                                                       effects on patients’ metabolism and for the improvement
                                                                                  of the risk stratification of AF patients.
                    RISK STRATIFICATION IN PATIENTS WITH ATRIAL                   References:
                    FIBRILLATION IN TREATMENT WITH ORAL                            1. Kornej, J., Börschel, C. S., Benjamin, E. J. &
                    ANTICOAGULANTS: AN NMR-BASED                                      Schnabel, R. B. Epidemiology of Atrial Fibrillation
                    METABOLOMICS AND LIPOPROTEOMICS STUDY                             in the 21st Century. Circulation Research 127, 4-20
                                                                                      (2020).
                    A. Vignoli1,2, F. Cesari3, L. Tenori1,2, E. Salvadori4,        2. Lau, Y. C. & Lip, G. Y. H. Which drug should we
                    B. Formelli5, A. Kura3, A.M. Gori3, B. Giusti3,                   use for stroke prevention in atrial fibrillation? Curr
                    R. Marcucci3, A. Poggesi5                                         Opin Cardiol 29, 293-300 (2014).




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                     3. Lip, G. Y. H. & Lane, D. A. Stroke prevention in          cient plasma used in the assay on the accuracy of the
                        atrial fibrillation: a systematic review. JAMA 313,       results. The use of porcine standard (when available)
                        1950-1962 (2015).                                         and FVIII deficient plasma with vWF is recommended.
                     4. Poggesi, A. et al. Role of Biological Markers for
                        Cerebral Bleeding Risk STRATification in Patients         Table 1.
                        with Atrial Fibrillation on Oral Anticoagulants for
                        Primary or Secondary Prevention of Ischemic
                        Stroke (Strat-AF Study): Study Design and
                        Methodology. Medicina (Kaunas) 55, E626 (2019).
                     5. Vignoli, A. et al. High-Throughput Metabolomics
                        by 1D NMR. Angew. Chem. Int. Ed. Engl. 58, 968-
                        994 (2019).


                    PO058

                    ANALYTICAL PERFORMANCE OF DIFFERENT
                    LABORATORY METHODS FOR MEASURING




                                                                                                         ly
                    SUSOCTOG-ALPHA




                                                                                                 on
                    C. Novembrino1, I. Quaglia2, M. Boscolo-Anzoletti1,
                    E. Galbiati1, A. Valpreda3
                                                                                  PO059




                                                                                             e
                    1Fondazione   IRCCS Ca’ Granda Ospedale Maggiore
                    Policlinico, Angelo Bianchi Bonomi Hemophilia and
                    Thrombosis Center, Milan, Italy; 2Istituto Clinico
                    Humanitas, Milan, Italy; 3Azienda Ospedaliera Città
                                                                                      us
                                                                                  A COMPARISON BETWEEN NON-ACTIVATED
                                                                                  THROMBOELASTOMETRY AND THROMBIN
                                                                                  GENERATION ASSAY FOR EMICIZUMAB
                    della Salute e della Scienza, Turin, Italy
                                                                                  al
                                                                                  MONITORING
                                                                          ci

                    Background and Aims: Acquired haemophilia A                   S. Arcudi1, E. Scalambrino2, M. Clerici2,
                    (AHA) is an often severe bleeding disorder caused by          C. Novembrino2, A. Tripodi2, F. Peyvandi1
                                                                    er


                    the development of antibodies (inhibitors) to factor VIII
                    (FVIII). Recombinant porcine factor VIII (rpFVIII,            1Department of Pathophysiology and Transplantation,
                                                              m



                    susoctocog alfa, Obizur®, Baxalta US Inc.-Takeda com-         Università degli Studi di Milano and Angelo Bianchi
                    pany- Bannockburn, IL, USA) is indicated for the treat-       Bonomi Hemophilia and Thrombosis Center and
                                                  om




                    ment of acute bleeding episodes in AHA, but there are         Fondazione Luigi Villa, Fondazione IRCCS Cà Granda
                    few data regarding the laboratory methods for properly        Ospedale Maggiore Policlinico, Milan, Italy; 2Angelo
                    monitoring the treatment. This study involving 3 differ-      Bianchi Bonomi Hemophilia and Thrombosis Center and
                    ent laboratories aimed to evaluate the analytical per-
                                               -c




                                                                                  Fondazione Luigi Villa, Fondazione IRCCS Cà Granda
                    formance of different assays for measuring rpFVIII.           Ospedale Maggiore Policlinico, Milan, Italy
                                       on




                    Methods: Five spiked samples at different rpFVIII con-
                    centrations (from 0.05 U/mL to 1.5 U/mL) were                 Background and Aims: The ability of coagulation
                    analysed on three distinct days, in triplicate every day,     assays to monitor severe hemophilia A (HA) patients on
                                  N




                    with nine different combinations of reagents (SynthasIL       emicizumab prophylaxis has been poorly investigated.
                    and SynthaFax -Werfen- for one-stage assay,                   Given this background, our study aims to report the
                    Chromogenix Coamatic FVIII for chromogenic assay),            results of non-activated rotational thromboelastometry
                    FVIII depleted plasmas (with or without von Willebrand        (NATEM) and thrombin generation assay (TGA) per-
                    factor –vWF-) and calibrators (HemosIL human calibra-         formed in patients on emicizumab prophylaxis and to
                    tor plasma, porcine calibrator diluted in FVIII deficient     correlate findings with plasma emicizumab concentra-
                    plasma with or without vWF). All the assays were per-         tion (EmiC). Methods: Consecutive HA patients on emi-
                    formed on ACL Top analysers (Werfen, Bedford, MA).            cizumab prophylaxis at steady state were enrolled in an
                    Intra- and inter assay and inter-laboratory Coefficient of    observational prospective study. TGA was assessed with
                    Variation (CV%) were calculate together with percent-         a homemade method upon addition to plasma of tissue-
                    age of recovery (% recovery) on the expected value.           factor (1pM) and synthetic phospholipids (1µM).
                    Results: The results of the three laboratories are report-    NATEM was measured for whole blood after addition of
                    ed in Table 1 as total inter-laboratory CV% (mean of          CaCl2 (100mM). Emicizumab plasma concentrations
                    CV% obtained for all the measures of the 5 samples in         (EmiC) were measured by the modified calibrated one
                    the three laboratories) and % recovery (mean of %             stage FVIII assay. We considered for TGA lag-time (LT),
                    recovery obtained for all the measures of the 5 samples       endogenous thrombin potential (ETP) and peak-throm-
                    in the three laboratories). Conclusions: The present          bin (PH); for NATEM, clotting-time (CT), clotting-firm-
                    study highlights the impact of the calibrator and the defi-   ness-time (CFT), time-to-maximum-clotting-firmness




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 120



         Poster




                    (MCF-t). Spearman’s Rho coefficient was used to assess        weight multimers (HMWM). Ristocetin-induced platelet
                    the correlation between variables. Results: Sixteen           agglutination assay (RIPA) or genetic analysis of von
                    patients on emicizumab prophylaxis (aged 17-64 years)         Willebrand factor (VWF) exon 28 is then performed to
                    were included in the study. Three were positive for FVIII     confirm diagnosis. Type 2B with HMWM (e.g.,
                    inhibitors. Results for NATEM and TGA are reported in         p.Pro1266Leu) might be missed at the first screening or
                    Table 1. When assessing the correlation with EmiC,            misdiagnosed as type 1/low-VWF and RIPA or genetic
                    NATEM-CT, NATEM-CT+CFT and NATEM-MCF-t                        analysis are rarely performed in these cases. Due to the
                    were inversely correlated with EmiC (CT rho=-0.536,           relative common presence of type 2B with HMWM vari-
                    p<0.05; CT+CFT rho=-0.515, p<0.05; MCF-t rho=-                ants in our type 2B population (25%), we developed a
                    0.561, p<0.05). TGA-ETP and TGA-PH were positively            screening assay to identify the type 2B variants using
                    correlated with EmiC (ETP rho=0.507, p<0.05; TGA-PH           patients’ frozen plasma. Methods: A wild-type recombi-
                    rho=0.643, p<0.01). Moreover, TGA and NATEM assays            nant glycoprotein Ib (rGPIb) was immobilized on a 96-
                    resulted to be inversely correlated (CT vs ETP, rho=-         wells ELISA plate using the 2D4 monoclonal antibody. A
                    0.771, p<0.001; CFT vs ETP, rho=-0.765, p<0.001;              normal pooled plasma (NP), type 2B controls and
                    CT+CFT vs ETP rho=-0.805, p<0.001). Conclusions:              patients’ plasma were normalized to the lowest VWF anti-
                    Our analysis of 16 patients on emicizumab prophylaxis         gen level. Each sample was seeded in presence of risto-
                    shows that both assays, NATEM and TGA, correlate with         cetin at 2 different concentrations (0.06 and 0.12 mg/mL;
                    the plasma concentration of emicizumab. Dose-adjust-          Figure 1) and incubated for 2 hours. VWF was detected
                    ment of emicizumab based on laboratory testing is cur-        with an anti-VWF HRP–labeled polyclonal antibody.




                                                                                                           ly
                    rently not required routinely. However, our preliminary       Seventy patients with VWF levels borderline or reduced,
                    data suggest that NATEM and TGA might be useful to




                                                                                                   on
                                                                                  at their first clinical evaluation due to bleeding diathesis,
                    help managing special situations requiring monitoring         were assessed versus a group of 14 type 2B patients.
                    with a global test, such as surgeries or concomitant treat-   Sanger sequencing of exon 28 was performed in patients
                    ment with bypassing agents.                                   with VWF increased binding to rGPIb. Results: We cal-




                                                                                              e
                                                                                  culated the mean values of the optical density (OD) ratios
                    Table 1.
                                                                                      us
                                                                                  between patient ‘s samples and NP, in comparison with
                                                                                  those obtained with the type 2B patients. Considering ris-
                                                                                  tocetin 0.06 mg/mL concentration, patients with VWF
                                                                                  al
                                                                                  increased affinity to GPIb shown a mean OD ratio higher
                                                                                  or equal to 1.87±0.70 versus mean OD ratio higher or
                                                                          ci

                                                                                  equal to 0.69±0.14 in patients with VWF normal affinity
                                                                                  to GPIb. Conclusions: This GPIb ELISA assay, that has
                                                                    er


                                                                                  the advantage to use frozen plasma, could help to identify
                                                                                  type 2B variants and can be used to select patients to be
                                                              m



                                                                                  investigated at molecular level.
                                                  om
                                               -c
                                       on




                      PO060
                                  N




                    A SCREENING ASSAY FOR TYPE 2B VON
                    WILLEBRAND FACTOR VARIANTS USING AN
                    ELISA ASSAY WITH RISTOCETIN AND A
                    RECOMBINANT WILD-TYPE GLYCOPROTEIN IB

                    P. Colpani1, L. Baronciani1, G. Cozzi1, M.T. Pagliari2,
                    A. Cairo1, E. Biguzzi1, F. Peyvandi1,2                        Figure 1.
                    1Fondazione IRCCS Ca’Granda Ospedale Maggiore

                    Policlinico, Angelo Bianchi Bonomi Hemophilia and             PO061
                    Thrombosis Center and Fondazione Luigi Villa, Milan,
                    Italy; 2Department of Pathophysiology and
                    Transplantation, Università degli Studi di Milano, Milan,     POPULATION PHARMACOKINETICS OF
                    Italy                                                         DIFFERENT CLOTTING FACTOR CONCENTRATES
                                                                                  IN FIFTY-SIX PATIENTS WITH SEVERE
                    Background and Aim: Type 2B patients are usually              HEMOPHILIA A: A SINGLE CENTER EXPERIENCE
                    detected using the first level assays for von Willebrand
                    disease (VWD), because of their loss of high molecular        S. Pasca1, E. Zanon2




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                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    1Biomedical  Sciences Department (DSB), Padua                  sometimes different from those obtained in the pivotal
                    University Hospital; 2Hemophilia Center, General               studies, therefore before starting a prophylaxis with any
                    Medicine, Padua University Hospital, Italy                     clotting factor concentrates, it is very useful to carry out
                                                                                   a PK evaluation, which together with an appropriate
                    Background and Aims: There are currently many factor           medical history, can guarantee the patient an adequate
                    VIII (FVIII) concentrates, either standard or extended         treatment for his needs and lifestyle.
                    half-life, available to clinicians that can certainly
                    improve the quality of life of patients with hemophilia A.
                    However, given the great inter and intra-individual vari-
                    ability, a correct pharmacokinetic evaluation is needed to
                    better find the most suitable drug for each patient.
                    Population pharmacokinetics (PopPK) make it possible
                    to easily obtain these data using a few samples and in a
                    short time. The aim of our study was to compare the PK
                    results obtained with different FVIII concentrates in dif-
                    ferent patients with hemophilia A. Methods: We have
                    analyzed, described, and compared all PopPK profiles of
                    different FVIII concentrates, standard (including recom-
                    binant FVIII single chain – rFVIII-sc) and extended half-




                                                                                                            ly
                    life, obtained in a group of severe hemophilia A patients
                    using the Web-Accessible Population Pharmacokinetics




                                                                                                    on
                    Service (WAPPS). For all products FVIII activity was
                    assessed using a one-stage assay (OSA) with ellagic acid
                    (Actin®FS). The rFVIII-sc values were multiplied by the




                                                                                               e
                    conversion factor 2.0. Results: Overall, we have collect-
                    ed 121 PopPK in 56 different patients with severe hemo-
                    philia A. 88 of which were obtained with standard half-
                    life (SHL) concentrates in 47 patients, while the remain-
                                                                                       us
                                                                                   al
                    ing 33 with extended half-life (EHL) products in 26 sub-
                    jects. 17 patients had both evaluations and switched from
                                                                           ci

                    SHLs to EHLs after PopPK assessment. Mean age at the
                    time of PopPK was similar between SHL and EHL
                                                                    er


                    groups, 23 years vs 26 years; and the mean dose of con-
                    centrate used in the two groups was 34.3 IU/kg and 35.0
                                                               m



                    IU/kg, respectively. Mean half-life was 1.38-fold higher
                    (Figure 1-c) with EHL products (16.8 hours) than SHL
                                                  om




                    (12.2 hours), with many differences between the differ-
                    ent FVIII concentrates analyzed. Figures 1-a and 1-b
                    show the various half-lives obtained with the different
                                               -c




                    concentrates, respectively with SHLs and EHLs. The 17
                    patients who underwent the switch had an improvement
                                       on




                    in the mean half-life of the drugs used which went from
                    12.5 to 17.1 hours, allowing them to reduce their infu-
                    sions by a mean of one third. No significant differences
                                  N




                    in blood group or in mean vWF:Ag/vWF:RCo were
                    observed between the different two groups, SHLs vs
                                                                                   Figure 1.
                    EHLs, or among patients treated with the different drugs.
                    Conclusions: Our study comprising over a hundred PK
                    analyzes, performed in 56 different patients, showed a
                    great PK variability existing between drugs. The OSA           PO062
                    used to evaluate PK profiles is usually considered accept-
                    able for all the tested drugs, therefore it did not signifi-   ANALITYCAL EVALUATION OF PRO II AND
                    cantly influence the data we obtained, although in any         PLATFORM POCT
                    case, great attention must always be paid to the assays
                    and reagents used when deciding to perform a PK assess-        C. Dellanoce, A. Zerbi, C. Zecca, N. Portesi,
                    ment. rFVIII-sc showed a mean half-life very close to          M. Di Gioia, O. Paoletti, R. Morandini, M. Tala, S. Testa
                    damoctocog-alfa, despite not being considered an EHL.
                    Among the other SHLs, the best result in terms of half-        Haemostasis and Thrombosis Center; Department of
                    life was obtained from the only tested plasma-derived          Laboratory Medicine, ASST Cremona, Italy
                    FVIII, while among the EHLs turoctocog-alfa-pegol
                    emerges with a half-life 1.25 times higher than those of       Background and Aims: AVK therapy is monitored
                    the other drugs with extended half-life. These results are     through PT-INR to ensure the therapeutic range stability.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 122



         Poster




                    Portable coagulation systems are an easy and immediate       Background and Aims: Prothrombin Time (PT INR) is
                    methods to perform PT-INR test on capillary blood. Many      the test used for monitoring Anti-Vitamin K
                    differences due to reagents, coagulometers and calibra-      Antagonists (VKA). The detection of the Clot
                    tions can allow different INR results. Agreement between     Waveform Analysis (CWA) was introduced for study-
                    portable monitors and reference systems should be always     ing the dynamic process of clot formation. The aim of
                    evaluated before their routine use for patient manage-       this study was to measure the PT 1st and 2nd deriva-
                    ment. Aim of the study was evaluate two commercial           tives (D) in anticoagulated patients for achieve informa-
                    portable monitors, Pro II and Platform, in comparison        tion on the clotting process. Methods: We studied a
                    with PT-INR provided by central laboratory reference         total of 780 patients (426 men and 354 women, median
                    system. Methods: PT-INR was performed on two                 age 75 years, range 13-100). PT ratio (RecombiPlasTin
                    portable monitors: Coagucheck Pro II (Roche                  2G, (Werfen), was performed using an ACL TOP 500
                    Diagnostics, Basel, Switzerland) Platform (LumiraDx,         (Werfen) which uses a turbidimetric method for clot
                    UK, Limited) which both use recombinant human throm-         detection. CWA detected the time at which maximum
                    boplastin with ISI=1.00. The local reference system was      velocity of clot formation was reached (1st D, as
                    STAR Max2 (Stago, France) a magneto-mechanical ana-          mAbs/s), and the time at which maximum acceleration
                    lyzer with rabbit thromboplastin (NeoPTimal, Stago           of clot formation was reached (2nd D, as mAbs/s2).
                    France, ISI=1.04). 126 AVK patients, with a stable PT-       Results: PT INR was not different between men and
                    INR ranging from 2.0 to 4.0, were evaluated. In the same     women (2.63, 0.95-9.46 and 2.99, 1.01-11.05 respec-
                    daily session venous blood samples were tested with the      tively, p=0.356) while a difference was found between




                                                                                                         ly
                    reference system and capillary PT-INR test was per-          both derivatives of men and women: 1st D 203.38,
                    formed on the two portable monitors. Linear regression




                                                                                                 on
                                                                                 45.24-693.57 and 223.57, 28.51-622.17 mAbs/s respec-
                    and Bland&Altman method were used for the statistical        tively (p<0.0001) and 2nd D (752.18, 161.17-5359.55
                    analysis. Results: The correlation among different sys-      and 818.68, 77.53-5049.58 mAbs/s2 respectively
                    tems was good. Coefficient of regression (R²) of             (p<0.006). A nonlinear regression showed a significant




                                                                                             e
                    NeoPTimal vs Pro II=0.914 and vs Platform=0.875; Pro II      correlation between PT INR and both 1st DER
                    vs Platform=0.844 (see Table 1). Differences in PT
                    INR≥±0.5 were observed in about 12% of the total popu-
                    lation, specifically 10.3% between NeoPTimal and
                                                                                     us
                                                                                 (R2=0.339) and 2nd D (R2=0. 501). A trend to a plateau
                                                                                 is present from INR from 3.0 to above 4.00 for the 2nd
                                                                                 D (Figure 1). Conclusions: 1st and 2nd D highlights a
                                                                                 al
                    Platform, 3.96 % between NeoPTimal and Pro II, 3.96 %        more reactive clot formation in women. CWA also
                    between the two POCT. Bland&Altman method showed             shows that the deep of anticoagulation did not change
                                                                         ci

                    a bias as follow: NeoPTimal vs Coagucheck Pro II=-0.3        much for INR above 3.0.
                    while the same thromboplastin vs Platform=2.6; between
                                                                   er


                    the two POCT=2.6. Conclusions: Results showed good
                    correlation between the different methods. Pro II showed
                                                             m



                    a better correlation and agreement compared to the labo-
                    ratory reference system. These differences could be due to
                                                 om




                    different thromboplastins, calibrations and reference sys-
                    tems. It is advisable that each center performs analytical
                    evaluations before introducing POCT in clinical practice.
                                              -c




                    Table 1.
                                       on
                                  N




                                                                                 Figure 1.



                      PO063                                                      PO064

                    CLOT WAVE ANALYSIS OF PROTHROMBIN TIME                       EVALUATION OF A POCT SYSTEM FOR TESTING
                    DURING ANTI-VITAMIN K ANTAGONISTS                            D-DIMER IN THE EXCLUSION OF
                                                                                 THROMBOEMBOLIC EVENTS
                    M.F. Ruberto1, F. Marongiu1, A. Mameli2, P. Schirru2,
                    S. Cornacchini2, D. Barcellona1                              A. Fassina, L. De Valentin, S. Luzi, R. Turrini, K. Proko,
                                                                                 L. Zardo
                    1Department   of Medical Sciences and Public Health,
                    University of Cagliari; 2Azienda Ospedaliero-                Laboratorio Analisi, ULSS 2, Marca Trevigiana, Treviso,
                    Universitaria di Cagliari, Italy                             Italy




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                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Background and Aims: D-Dimer is a protein fragment           atives occur in these cases too. In conclusion, POCT
                    detectable within 4-6 hours in blood, from the begin-        system is a valid alternative to the laboratory method, in
                    ning of the fibrinolysis process. D-Dimer test is an inte-   particularly for clear positive or negative results. For
                    gral part of validated algorithms for the diagnosis of       borderline results, confirmation by laboratory testing
                    deep-vein thrombosis (DVT) and pulmonary embolism            may be suggested.
                    (PE) avoiding invasive and expensive imaging tests.
                    The importance of D-Dimer test during COVID-19
                    pandemic is testified by the doubling of D-Dimer             PO065
                    requests. The aim of this study is to evaluate the per-
                    formance of a Point of Care Testing (POCT) for D-
                                                                                 PROTHROMBIN TIME (PT) AND ACTIVATED
                    Dimer determination compared to the laboratory assay.
                    Methods: The D-Dimer plasma concentration, obtained          PARTIAL THROMBOPLASTIN TIME (APTT)
                    from POCT (microfluidic immunofluorescent,                   TESTING: RAPID CENTRIFUGATION VERSUS
                    LumiraDx), was compared to the one resulted from the         CLASSICAL CENTRIFUGATION
                    laboratory method (turbidimeter, ACL-TOP, Werfen).
                    The results were represented in Bland-Altman Plot. 105       M. Gagliardi1, L. Capone2, P. Migliaccio2,
                    samples from Emergency Room (E.R.), hospital units           M.R. Cianciulli2, V. Marottoli3, A. Ciampa1
                    and external patients were analyzed for D-Dimer assay
                                                                                 1Haemostasis   Centre, S.GMoscati Hospital, Avellino;
                    on the reference instrument ranging from 184 to 4965




                                                                                                         ly
                                                                                 2Laboratory of Clinical Pathology, S.GMoscati Hospital,
                    ng/mL FEU. The evaluation was performed using sodi-
                    um citrate plasma samples collected for D-Dimer assay,       Avellino; 3Multimedica SrL, Napoli, and Mediterranea




                                                                                                 on
                    firstly on the laboratory instrument and then on POCT,       Diagnostica, Castellammare di Stabia (NA), Italy
                    within approximately 4 hours. Results: 37 of the 105
                    samples tested were negative with the reference method       Background and Aims: Prothrombin time (PT), and




                                                                                            e
                    and 68 were positive. Age-modified cut-off (500 ng/mL        activated partial thromboplastin time (APTT) are basic
                    FEU up to 50 years and 10 ng/mL more for each year of
                    age) was used, as recommended by qualified publica-
                    tions (Lippi et al., D-Dimer Testing for Suspected
                                                                                     us
                                                                                 tests performed for coagulation screening disorders,
                                                                                 and for monitoring patients on anticoagulant therapy. In
                                                                                 coagulation tests, sample processing and preparation is
                                                                                 one of the most critical steps affecting reporting times.
                                                                                 al
                    Venous Thromboembolism in the Emergency
                    Department, Clin Chem Lab Med 2014), to define pos-          Centrifugation is the pre-analytical phase that most lim-
                                                                         ci

                    itive or negative results. The agreement between refer-      its speed in sample processing. The aim of this work is
                    ence method and POCT was 88.6%, considering posi-            to demonstrate that it is possible to reduce the reporting
                                                                   er


                    tive-positive and negative-negative. POCT results evi-       time for coagulation tests by using a centrifugation with
                    denced 4 out of 105 false negatives and 8 out of 105         a higher centrifugal force and a lower rotation time that
                                                             m



                    false positives. In all the 4 false negative cases D-Dimer   one recommended by Laboratory Guidelines, obtaining
                    determinations were just below the age-adapted cut-off       a qualitatively suitable plasma. Methods and Patients:
                                                 om




                    and with small quantitative differences compared to the      Three hundred samples obtained from outpatients and
                    reference method. Sensitivity and specificity were           inpatients for PT and APTT estimation were included.
                    respectively 94.1% and 78.4%. Bland Altman Plot is           Overfilling samples, insufficient and lipemic filling
                                              -c




                    used to evaluate methods’ agreement. The difference          samples were excluded. Tube 1 was centrifuged in a
                    mean line is very close to zero (44 ng/mL), so there is      benchtop centrifuge (JOUAN B4i) at 1500 g for 20
                                       on




                    no clear tendency to underestimate or overestimate. In       minutes. Tube 2 was centrifuged with the same cen-
                    the Bland-Altman Plot points near the age-modified           trifuge at 3026 g for 4 minutes. No brakes were used.
                    cut-off are very close to the mean line showing mini-        The plasma obtained with the two methods was tested
                                  N




                    mum differences between methods; whereas, for con-           for PT and APTT using the automated method on the
                    centrations >500 ng/mL, points are far from the mean         ACL TOP 500 coagulometer (Werfen). To confirm the
                    line. Disagreement results only emerged near the age-        adequacy of platelet-poor plasma, platelet counts were
                    modified cut-off where the patient classification is dif-    performed on plasma obtained from 40 random samples
                    ficult using only D-Dimer results, so any important          out of 300 samples included. Residual platelets were
                    clinical impact can be excluded. Conclusions: Our            evaluated in accordance with CLSI recommendations,
                    evaluation shows a good agreement of POCT system             and the sample test results were less than 10 000/μl.
                    and reference method. POCT systems are particularly          Platelet count was performed on the automated hemato-
                    useful in E.R. and situations in which using laboratory      logical analyzer COULTER LH 780. A routine internal
                    test is difficult. In these contexts, the use of POCT is     quality control for the analyzer including platelet count
                    important in order to reduce Turn Around Time and            was performed daily, using BIORAD three-level con-
                    help early decision making for patient management. In        trols. Results: PT and APTT results obtained by con-
                    the determination of D-Dimer, high sensitivity is essen-     ventional centrifugation and rapid centrifugation were
                    tial to avoid false negative results, so that DVT or PE      compared. A comparison was also made among INR
                    potentially patients can be correctly screened.              values that were subtherapeutic (<2.0), INR values
                    Considering POCT as an exclusion test, its performance       within the therapeutic range (between 2.0 and 3.0), and
                    should be evaluated in selected patients already diag-       that ones above the therapeutic range (INR >3.0). The
                    nosed for DVT or PE, just to verify whether false neg-       data were analyzed using descriptive statistics, Student




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 124



         Poster




                    t-tests, correlation coefficient (r) and Bland-Altman          hood. CAMT is mostly caused by mutations in MPL,
                    graphs for comparison of methods (Figure 1). The mean          the gene encoding the receptor of thrombopoietin
                    values for PT and APTT for both conventional and               (THPO), a crucial cytokine regulating hematopoiesis.
                    rapid centrifugation were comparable, without statisti-        CAMT can be also due to mutations affecting the
                    cally significant differences. Conclusions: The present        THPO coding region. In a child with CAMT clinical
                    study has shown that by rapid centrifugation of sam-           picture, we identified the homozygous c.-323C>T sub-
                    ples, 3026 g for 4 minutes, it is possible to obtain PT        stitution, affecting a potential regulatory region of
                    and APTT values, almost identical to those values              THPO. Though mechanisms controlling the THPO
                    obtained by conventional centrifugation (1500 g for 20         transcription are not characterized, bioinformatics and
                    minutes). No statistically or clinically significant differ-   in vitro analysis showed that c.-323C>T prevents the
                    ence between PT and APTT values was observed for               binding of transcription factors ETS1 and STAT4 to the
                    either type of centrifugation. This means that reporting       putative THPO promoter, impairing THPO expression.
                    time for routine PT/APTT tests can be reduced of 16            Accordingly, in the proband the serum THPO concen-
                    minutes: this is a significant time reduction in perform-      tration indicates defective THPO production. Based on
                    ing clotting tests, and it can have a positive impact on       these findings, the patient was treated with the THPO-
                    diagnostic and therapeutic decisions in patients care,         mimetic agent eltrombopag, inducing a significant
                    particularly in those patients hospitalized in intensive       increase in platelet count and stable remission of bleed-
                    care settings such as trauma units and operating the-          ing symptoms. Herein, we report a novel pathogenic
                    atres. This will also reduce the waiting time of outpa-        variant responsible for CAMT and provide new insights




                                                                                                           ly
                    tients for monitoring oral anticoagulant therapy.              into the mechanisms regulating the transcription of the




                                                                                                   on
                                                                                   THPO gene.


                                                                                   PO067




                                                                                              e
                                                                                       us
                                                                                   MRP-4 (MULTIDRUG RESISTANCE PROTEIN-4)
                                                                                   PROTEIN EXPRESSION IN PATIENTS WITH ACUTE
                                                                                   CORONARY SYNDROME UNDER CHRONIC
                                                                                   al
                                                                                   ASPIRIN TREATMENT
                                                                           ci

                                                                                   E. Schiera1, M. Curreli1, V. Cesario2,
                                                                     er


                                                                                   F. De Felice2, G. Tocci3, F.M. Pulcinelli1
                                                               m



                                                                                   1Dipartimento di Medicina Sperimentale Sapienza

                    Figure 1.                                                      Università di Roma; 2UOC Cardiologia Interventistica,
                                                  om




                                                                                   AOSCamillo Forlanini; 3Dipartimento di Medicina
                                                                                   Clinica e Molecolare Sapienza Università di Roma, Italy
                      PO066
                                               -c




                                                                                   Background and Aims: Aspirin reduces cardiovascular
                                                                                   events. However, some patients under chronic aspirin
                    DEFECTIVE BINDING OF ETS1 AND STAT4 TO
                                       on




                                                                                   treatment present «High-on Aspirin Residual Platelet
                    THPO PROMOTER IN CONGENITAL                                    Reactivity» (HARPR) phenomenon. Platelet MRP-4
                    AMEGAKARYOCYTIC THROMBOCYTOPENIA                               overexpression correlate with HARPR, suggesting that
                                  N




                                                                                   MRP-4 overxpression is an additional CV risk factor.
                    V. Capaci1, E. Adam2, I. Bar-Joseph3, M. Faleschini1,          To evaluate platelets over-expression as an additional
                    A. Pecci4,5, A. Savoia1,6                                      CV risk factor, we performed a study in which we
                                                                                   investigated amounts of platelets MRP4 between the
                    1Institute
                             for Maternal and Child Health, IRCCS Burlo            ACS population and patients under chronic aspirin
                    Garofolo, Trieste, Italy; 2Department of Pediatric             treatment without ACS population. Methods: We ana-
                    Hematology, Oncology and Bone Marrow Transplant,               lyzed platelet protein expression in patients diagnosed
                    Sheba Medical Center, Tel Hashomer, Israel; 3The               with ACS on chronic aspirin therapy (100 mg/die) for at
                    Center for Cancer Research, Sheba Medical Center, Tel          least two months (SCA) and patients on chronic aspirin
                    Hashomer, Israel; 4Department of Internal Medicine,            therapy (100 mg/die) for at least two months, in the
                    IRCCS Policlinico San Matteo Foundation, Pavia, Italy;         absence of ACS (ASA). Between 1st December 2021
                    5University of Pavia, Pavia, Italy; 6Department of             and 1st march 2022 we enroled patients with ACS on
                    Medical Sciences, University of Trieste, Trieste, Italy        chronic aspirin therapy (100 mg/die) for at least two
                                                                                   months (N=51)) and patients on chronic aspirin therapy
                    Congenital amegakaryocytic thrombocytopenia                    (100 mg/die) for at least two months, in the absence of
                    (CAMT) is a recessive disorder characterized by severe         ACS (ASA; N=30) from San Camillo Interventional
                    reduction of megakaryocytes and platelets at birth,            Cardiology Unit and Platelet physiopatology Lab,
                    which evolves toward bone marrow aplasia in child-             Policlinico Umberto I respectively. Levels of platelets




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 125



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    MRP4 expression were evaluated with Western Blot            occasionally required. Thrombin generation assay
                    analysis comparing the densitometric values of protein      (TGA) can be used to monitor the global hemostatic
                    expression of each individual belonging ACS and ASA         response in emicizumab treated patients. Aims. To
                    populations. Data were expressed as percentage of the       measure emicizumab plasma levels and TGA in severe
                    densitometric mean (ImageJ, NIH) of each sample             hemophilia A (HA) patients without and with inhibitors.
                    compared to the average of the densitometric values         Methods: A modified one-stage FVIII clotting assay
                    obtained from three healthy volunteers (HV). Each gel       has been used to measure emicizumab concentration.
                    data were normalized using the same HV samples.             The assay is calibrated against emicizumab using a ded-
                    Results: In platelets obtained from ACS patients we         icated plasma emicizumab calibrator and two levels of
                    found a higher amount of MRP4 platelets expression          control. Thrombin generation was determined by
                    (50% higher) compared to those obtained in ASA              Calibrated Automated Thrombography (CAT) using
                    patients. In Figure 1 is reported a representative case     PPP-Reagent (phospholipids [4 uM], TF [5 pM]).
                    that show a higher platelet MRP4 level vs ASA patients      Results: 10 patients (non-inhibitor 6; inhibitor 4)
                    in a patient with acute ST segment elevation myocardial     received emicizumab. At start, 6 were without and 4
                    infarction secondary to a thrombus that stopped one         with inhibitor. No spontaneous bleedings occurred. Two
                    week before double antiplatelet treatment.                  patients experienced traumatic bleedings. One was
                    Conclusions: Reduced sensitivity to aspirin by MRP-4        treated with rFVIII concentrate (14th dose of life) and
                    could be one of the causes of higher risk for SCA           subsequently developed a high titer inhibitor. At the
                    events. In the context of precision medicine platelet       time of blood drawn he was considered an inhibitor




                                                                                                        ly
                    MRP4 levels may be a useful tool to identify patients       patient. Emicizumab plasma levels increase during
                                                                                loading phase; median concentration at steady state is




                                                                                                on
                    less sensible to aspirin treatment and at high CV risk
                    factor. In addition it could be important to identify       42.2µg/mL (25.3-75). Lag time is in normal range after
                    patients that required antiplatelet treatment variation     the first dose; ETP is variable among patients, almost
                    adding MRP4 inhibitor to aspirin treatment, so as to        always below normal range, and does not significantly




                                                                                           e
                    create a personalized treatment in order to reduced CV      differ between loading and steady state phase (Table 1).
                    complications.
                                                                                    us
                                                                                Comparing non inhibitor and inhibitor patients, at
                                                                                steady state median emicizumab concentration is 58.4
                                                                                µg/mL (46.1-70.7) and 34.7 µg/mL (25.3-75), respec-
                                                                                tively. In non-inhibitor patients, median ETP and peak
                                                                              al
                                                                                are higher than in inhibitor patients: ETP 1073.14
                                                                          ci

                                                                                nMxmin (562.17-1412.23), peak 92.67 nM (38.43-
                                                                                107.11) versus ETP 688.64 nMxmin (369.64-895.24),
                                                                  er


                                                                                peak 43.065 nM (22.99-56.66). Conclusions: We
                                                                                observed variability among patients as regards emi-
                                                            m



                                                                                cizumab concentration and TGA, but the drug was effi-
                                                                                cacious in all. It has to be explored if variables such as
                                                 om




                                                                                inhibitors, BMI, patients age, adherence to treatment,
                                                                                time period from the last infusion, influences emi-
                                                                                cizumab concentration and/or TGA.
                                              -c




                                                                                Table 1.
                                      on




                    Figure 1.
                                  N




                    PO068

                    EVALUATION OF EMICIZUMAB CONCENTRATION
                    AND THROMBIN GENERATION IN A COHORT OF
                    SEVERE HEMOPHILIA A PATIENTS WITH AND
                    WITHOUT INHIBITOR DURING EMICIZUMAB
                    PROPHYLAXIS                                                 PO069
                    M. Mormile, E. Baldacci, A. Ferretti, G. Maestrini,
                    F. Viola, A. Chistolini, C. Santoro                         EVALUATION OF THE ROLE OF NEW
                                                                                LABORATORY PARAMETERS IN THE
                    Dipartimento di Medicina Traslazionale e di Precisione,     MANAGEMENT AND THE PREVENTION OF
                    Istituto di Ematologia, Università “La Sapienza”, Roma,     RELAPSES IN PATIENTS WITH ACQUIRED TTP
                    Italy                                                       TREATED WITH CAPLACIZUMAB AND ANTI-CD20

                    Background and Aims: Although emicizumab is                 R. Tomasino1, M. Napolitano3, F. Bonura1,2, F. Schiralli1,
                    administered at fixed dose, its measurement could be        E. Zora1, M. Ciaccio1,2




                                                             [Hematology Reports 2021; 13:s3]                                            [page 125]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 126



         Poster




                    1Department    of Laboratory Medicine, University            have been subject to several relapses close in time. This
                    Hospital “Paolo Giaccone”, Palermo, Italy;                   could be due to the delay in the introduction of the
                    2Department of Biomedicine, Neurosciences and                Rituximab and the laboratory parameters may be able to
                    Advanced Diagnostics, Institute of Clinical                  give an objective explanation for the fact that the only
                    Biochemistry, Clinical Molecular Medicine and Clinical       therapy with PEX+Caplacizumab is not effective for
                    Laboratory Medicine, University of Palermo, Palermo,         prevent the relapse. The course of the disease in the
                    Italy; 3Department of Haematology, University Hospital       other three patients would confirm the hypothesis that
                    “Paolo Giaccone”, Palermo, Italy                             the closest administration overtime of anti-CD20 to
                                                                                 Caplacizumab favors a complete remission and does
                    Background: Acquired Thrombotic Thrombocytopenic             not predispose to further relapses, at least in the short
                    Purpura (aTTP) is an acute, severe, rare syndrome of         term. The preliminary data described show that the new
                    difficult diagnosis related to antibodies anti-              laboratory parameters are configured as excellent can-
                    ADAMTS13. In about one-third of patients, aTTP               didates to improve the clinical-diagnostic scores, mak-
                    relapses most often during the first two years. The aim      ing them more effective in preventing relapses.
                    of the current study was to evaluate, by clinical and lab-
                    oratory parameters, the response to the treatment of
                    relapsing forms of aTTP treated with Caplacizumab and        PO070
                    subsequent administration of anti-CD20 (Rituximab).
                    Case Report: A cohort of 5 patients affected by




                                                                                                          ly
                    relapsed aTTP was followed over a period of 18               DECOMPENSATED CIRRHOSIS WITH BACTERIAL
                    months. The response to treatment with Caplacizumab          INFECTIONS SHOWS A PECULIAR




                                                                                                  on
                    and anti-CD20 was assessed by laboratory parameters,         DERANGEMENT OF THE HEMOSTATIC BALANCE
                    including platelets count, ADAMTS13 Activity levels,
                    and titers of anti-ADAMTS13 antibodies, measured by          E. Campello1, A. Zanetto2, L. Spiezia1, C. Bulato1,




                                                                                            e
                    both chemiluminescence (CLIA) and immunoenzymat-             S. Gavasso1, G. Saggiorato1, S. Shalaby2, P. Burra2,
                                                                                 M. Senzolo2, P. Simioni1
                    ic techniques. The HemosIL AcuStar ADAMTS13
                    Activity Kit by IL Werfen was used for the evaluation
                    of the ADAMTS13 Activity in CLIA and titration of
                                                                                     us
                                                                                 1Department
                                                                                 2Department
                                                                                              of Medicine, University of Padova, Italy;
                                                                                                  of   Surgery,     Oncology,      and
                                                                                 al
                    neutralizing ADAMTS13 antibodies was performed by
                    applying the modified Bethesda Nijmegen protocol; for        Gastroenterology, University of Padova, Italy
                                                                         ci

                    the titration of total antibodies the ELISA technique
                    was used. The cohort consists of one patient at the clin-    Background and Aims: Bacterial infections (BIs) in
                                                                   er


                    ical onset and four patients with aTTP diagnosed in pre-     cirrhosis are associated with increased bleeding risk. To
                    vious years, relapsing, undergoing traditional therapies     assess the factors responsible for bleeding tendency in
                                                             m



                    (PEX, Rituximab and corticosteroids) plus                    BIs, we conducted a prospective study comparing all
                    Caplacizumab. All five patients received Caplacizumab        components of hemostasis (platelets, coagulation, and
                                                 om




                    during PEX and after its suspension, according to rec-       fibrinolysis) in hospitalized patients with decompensat-
                    ommended indication. Three of them were also treated         ed cirrhosis with vs. without BIs. Methods: Primary
                    with anti-CD20 during hospitalization at the end of          hemostasis assessment included whole blood platelet
                                                                                 aggregation and von Willebrand factor (VWF).
                                              -c




                    PEX, while the other two received anti-CD20 in the fol-
                    lowing months during follow-up for ADAMTS13                  Coagulation assessment included procoagulant factors
                                       on




                    Activity below 20%. Conclusions: ADAMTS13                    (fibrinogen, factor II, V, VII, VIII, IX, X, XI, XII, XIII),
                    Activity is the cornerstone of the correct diagnostic        natural anticoagulants (protein C, protein S, antithrom-
                    framing of aTTP, but it does not seem to be a valid pre-     bin) and thrombomodulin-modified thrombin genera-
                                  N




                    dictive indicator of relapse. For this purpose, the anti-    tion test. Fibrinolysis assessment included fibrinolytic
                    body titer, inversely proportional to the ADAMTS13           factors (plasminogen, t-PA, PAI-1, α2-AP, TAFIa/ai)
                    Activity, may be a more useful tool. Bethesda method         and plasmin-antiplasmin complex (PAP). Results:
                    by CLIA to show the inhibitory function of anti-             Eighty patients with decompensated cirrhosis were
                    ADAMTS13 autoantibodies is important to confirm the          included (40 with and 40 without BIs). Severity of cir-
                    diagnosis of immune-mediated TTP, even in cases              rhosis and platelet count were comparable between
                    where the antibodies are not neutralising, since their       groups. At baseline, patients with cirrhosis and BIs had
                    presence does not necessarily exert an inhibitory effect     a significantly lower whole blood platelet aggregation,
                    on ADAMTS13 Activity. The determination of                   consistent with impaired platelet function, without sig-
                    ADAMTS13 Activity, however, is recommended dur-              nificant differences in VWF. Regarding coagulation,
                    ing follow-up to apply preventive therapeutic protocols.     BIs were associated with reduced procoagulant factors
                    Data have been useful to the clinician to monitor            VII and XII, and a marked reduction of all natural anti-
                    patients and to define the effectiveness of the treatments   coagulants. Thrombomodulin-modified thrombin gen-
                    administered, especially in cases of multiple relapses.      eration, however, was comparable between groups.
                    Specifically, for the two patients who have benefited        Finally, although mixed potentially hypo-fibrinolytic
                    from treatment with Rituximab only in the months fol-        (low plasminogen) and hyper-fibrinolytic (high t-PA)
                    lowing the treatment with PEX+Caplacizumab for var-          changes were present in BIs, comparable levels of PAP
                    ious reasons, one might venture the hypothesis that they     were detected in both groups. Upon resolution of infec-




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 127



                                                                             XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    tion (n=29/40), platelet aggregation further deteriorated         3Pediatric Ophthalmology Unit, AMeyer Children’s
                    whereas coagulation and fibrinolysis factors returned to          Hospital, Florence, Italy; 4Ophthalmology Unit,
                    levels observed in patients without BIs (Table 1).                Department of Neuroscience, University of Padua,
                    Conclusions: In hospitalized patients with decompen-              Padua, Italy; 5Fondazione IRCCS Ca’ Granda,
                    sated cirrhosis, BIs are associated with reduced whole            Ospedale Maggiore Policlinico, Angelo Bianchi Bonomi
                    blood platelet aggregation and a marked decrease of all           Hemophilia and Thrombosis Center, Milan, Italy; 6UOC
                    natural anticoagulants, which may unbalance hemosta-              Ophthalmology, Bambino Gesù Pediatric Hospital,
                    sis and potentially increase the risk of bleeding and             IRCCS, Rome, Italy; 7Medical Affairs Group, Kedrion
                    thrombosis.                                                       SpA, Lucca, Italy; 8Clinical Research Group, Kedrion
                                                                                      SpA, Lucca, Italy
                    Table 1. Hemostatic alterations in patients with cirrhosis with
                    vs. bacterial infections at baseline.                             Background and Aims: Congenital plasminogen
                                                                                      (PLG) deficiency is an ultrarare autosomal-recessive
                                                                                      systemic disorder caused by mutations in PLG gene that
                                                                                      affects ~1.6 million people worldwide. It is character-
                                                                                      ized by severely impaired extravascular fibrinolysis
                                                                                      leading to woody (ligneous) fibrin-rich pseudomembra-
                                                                                      nous formation on mucous membranes, such as eyelids,
                                                                                      respiratory tract, oropharynx, gums, middle ear, female




                                                                                                              ly
                                                                                      reproductive tract, renal collecting system, central nerv-
                                                                                      ous system and skin. Ligneous conjunctivitis (LC)




                                                                                                      on
                                                                                      appears to be the most common clinical manifestation
                                                                                      (80%) and presents with redness of the conjunctivae
                                                                                      and progress to pseudomembranes’ formation on the




                                                                                                   e
                                                                                      palpebral surfaces that replace the normal mucosa and

                                                                                          us
                                                                                      if untreated, can result in corneal involvement and relat-
                                                                                      ed damages with visual impairment or blindness.
                                                                                      Topical treatment with corticosteroids, cyclosporine,
                                                                                      al
                                                                                      azathioprine, and heparin have been used with limited
                                                                                      success; systemic Fresh Frozen Plasma was found to be
                                                                              ci

                                                                                      effective, but it is hampered by the need for repeated
                                                                                      infusions; topical plasminogen concentrate has demon-
                                                                       er


                                                                                      strated efficacy in several case studies. Six subjects are
                                                                                      currently on home treatment in Italy with human plas-
                                                                 m



                                                                                      ma-derived plasminogen eye drops (Ked-hPLG;
                                                                                      Kedrion Biopharma, Italy) to prevent recurrence of LC.
                                                    om




                                                                                      This study describes these 6 subjects’ clinical course
                                                                                      before and after use of Ked-hPLG eye drops. Methods:
                                                                                      Review of medical records of the 6 subjects with con-
                                                 -c




                                                                                      firmed diagnosis of LC at four centers, as per the
                                                                                      648/96 Law in force in Italy. These subjects are on
                                        on




                                                                                      home treatment with Ked-hPLG eye drops based on the
                                                                                      phase II/III clinical trial (KB046) results. Two subjects
                                                                                      came from the KB046 study. Results: All subjects are
                                   N




                                                                                      female and their age range at initiation of treatment
                                                                                      were 4-71 years (median 7.5 years). All subjects under-
                                                                                      went between 1-50 surgical interventions (median 5)
                                                                                      for removal, peeling, or debridement of pseudomem-
                    PO071
                                                                                      branes before initiating Ked-hPLG eye drops treatment.
                                                                                      These surgical procedures were followed by recur-
                    THE USE OF HUMAN PLASMINOGEN EYE DROPS                            rences of pseudomembranes in all 6 subjects despite
                    IN SUBJECTS AFFECTED BY LIGNEOUS                                  several topical treatments, including steroids, antibi-
                    CONJUNCTIVITIS IN ITALY                                           otics, heparin, cyclosporine and FFP. The treatment
                                                                                      with Ked-hPLG eye drops started with two drops for
                    M.T. Sartori1, A. Bartuli2, R. Caputo3, F. Cortellessa2,          each affected eye 4-8 times/day as loading dose and
                    A. Leonardi4, S.M. Siboni5, P. Valente6, L. Pino7,                then continued with a maintenance regimen of 2 drops
                    A. Lotti Suffredini8, P. Mathew7, R. Crea7                        per eye 4 times/day. Subjects’ follow up range is 22-141
                                                                                      months (median 102 months). The results, as shown in
                    1Clinical Medicine I, Department of Medicine, Padua               Table 1, provide evidence of the effectiveness of long-
                    University Hospital, Padua, Italy; 2UOC Rare Diseases             term treatment with no post-operative recurrence in all
                    and Medical Genetics, University Pediatric Dpt.,                  subjects on treatment with Ked-hPLG eye drops. No
                    Bambino Gesù Pediatric Hospital, IRCCS, Rome, Italy;              major safety concerns, including decrease in efficacy or




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 128



         Poster




                    hypersensitivity reactions, have been reported so far        of FXI to FXIa, which then activates FIX, Kallikrein
                    (Apr/2022) confirming the favorable safety and tolera-       (PKa), in a dose and time dependent manner, can direct-
                    bility profile of Ked-hPLG eye drops. The 2 subjects         ly activate FIX to generate FIXa, resulting in thrombin
                    from the KB046 study developed transient anti-apro-          generation and clot formation independently of FXIa.
                    tinin antibodies and 1 of them also developed transient      Methods: A 71-year-old male patient (1) was admitted
                    anti-PLG antibodies. However, none of them required          to our hospital to undergo a surgical procedure. The pre-
                    modification, interruption or discontinuation of treat-      operative screening showed a prolonged aPTT ratio of
                    ment. Data on the other 4 subjects are not available.        2,94 with a normal reference range of 0,85 to 1,20,
                    Conclusions: Congenital PLG deficiency is an ultrarare       while the PT ratio (0,99) was normal. Some years ago,
                    disease probably underdiagnosed requiring more atten-        he received a radical prostatectomy and some dental
                    tion to identify symptomatic subjects that might benefit     avulsions, without any bleeding. Another 71-year-old
                    from newer therapeutic options. Our study confirms           male patient (2) presented to our haemostasis laboratory
                    long term effectiveness from the use of Ked-hPLG eye         for a preoperative screening before undergoing a
                    drops, and is well tolerated and effective in preventing     prostatectomy. The aPTT was prolonged with a ratio of
                    pseudomembrane recurrence in subjects with LC.               2,73 while, also in this case, the PT ratio (0,90) was nor-
                                                                                 mal. His past medical history was negative for minor
                    Table 1.                                                     and major bleeding events. Two brothers of this
                                                                                 patient,(aPTT of 4,32 and 4,41)had a prekallikrein clot-
                                                                                 ting activity of 6% and 5%, respectively. Results: By




                                                                                                          ly
                                                                                 mixing an aliquot of normal plasma with plasma of
                                                                                 each patient in equal proportions, aPTTs normalized




                                                                                                  on
                                                                                 from 2,94 and 2,73 to 1,01 and 1,03, respectively.
                                                                                 Clotting activities of FXII, FXI, FIX, FVIII and an
                                                                                 immunoassay of vWFAg were normal. After a preincu-




                                                                                            e
                                                                                 bation of 30 m’ aPTTs normalized from 2,94 and 2,73

                                                                                     us
                                                                                 to 1,11 and 1,07, respectively. Patient (1) showed a
                                                                                 prekallikrein clotting activity of 2% and patient (2) of
                                                                                 9%. The sensitive reagents were colloidal silica and
                                                                                 synthetic phospholipids (Hemosil APTT-SP), with an
                                                                                 al
                                                                                 aPTT ratio of 2,94 patient (1) and of 2,73 patient (2),
                                                                         ci

                                                                                 and silicon dioxide and vegetables phospholipids
                                                                                 (Pathromtin SL) with an aPTT ratio of 4,2 (patient 1)
                                                                   er


                                                                                 and 3,8 (patient 2), which normalized to 1,25 and to
                                                                                 1,22, respectively, after 30 m’ of preincubation, while
                                                             m



                                                                                 aPTTs performed with ellagic acid and synthetic phos-
                                                                                 pholipids (Synthafax) were normal (Table 1).
                                                 om




                                                                                 Table 1.

                      PO072
                                              -c
                                       on




                    PREKALLIKREIN DEFICIENCY: DESCRIPTION OF
                    TWO CASES
                                   N




                    M. Gagliardi1, L. Capone2, A. Uva2, M.R. Cianciulli2,
                    V. Marottoli3, A. Ciampa1                                    Conclusions: About all patients with prekallikrein defi-
                                                                                 ciency have a personal and family history negative for
                    1Haemostasis  Centre, S.GMoscati Hospital, Avellino          bleeding events. Some arterial thrombotic events
                    2Clinical Pathology Laboratory, S.GMoscati Hospital,         observed in some patients seem related to the increased
                    Avellino; 3Multimedica SrL, Napoli, and Mediterranea         incidence of arterial hypertension and endothelial dys-
                    Diagnostica, Castellammare di Stabia (NA), Italy             function, due to lack of bradykinin formation, normally
                                                                                 released from High Molecular Weight (HMW)
                    Background: Prekallikrein (PK) is a zymogen that is          Kininogen by kallikrein (PKa). Bradykinin causes
                    converted to kallikrein (PKa) by FXIIa. Prekallikrein        vasodilation inducing release of powerful vasodilators
                    deficiency is a rare autosomal recessive defect caused       such as prostacyclin, endothelium-derived hyperpolariz-
                    by KLKB1 mutations, whose main characteristic is an          ing factor (EDHF) and nitric oxide (NO), which regu-
                    isolated prolongation of the aPTT, which becomes nor-        lates endothelial homeostasis too. Bradykinin causes also
                    mal after prolonged preincubation, due to autoactiva-        release of tissue plasminogen activator, that regulates fib-
                    tion of FXII. This deficiency is not associated with         rinolysis. Notably, because PK and FXII reciprocally
                    bleeding tendency. Even though PK and FXII recipro-          activate, and because PKa contributes to thrombin gener-
                    cally activate each other, and FXIIa initiates the intrin-   ation also through activation of FIX independently of
                    sic pathway of the coagulation system via the cleavage       FXII and FXI, the development of new anticoagulants,




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 129



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    which act as inhibitors of FXIIa may contribute to reduce     increased up to 38.3% after 14 days of combined therapy
                    PKa generation, and consequently to reduced FIXa with         with Susoctacog alfa and steroids. After 4 weeks, rfVIII
                    subsequent reduced thrombin formation.                        administration was interrupted and the patient was dis-
                                                                                  charged after 32 days of corticosteroid therapy with no
                                                                                  other spontaneous bleeding events, a factor VIII dosage
                    PO073                                                         of 20% and an inhibitor’s titer reduction. Conclusions:
                                                                                  AHA has been associated with several clinical condi-
                                                                                  tions. We found 13 Sars-CoV-2 mRNA vaccination cases
                    ACQUIRED HEMOPHILIA A IN A 74-YEAR-OLD
                                                                                  associated with the onset of AHA, occurring after a min-
                    WOMAN AFTER THE THIRD DOSE OF M-RNA
                                                                                  imum of 7 to a maximum of 52 days after the vaccina-
                    SARS-COV-2 VACCINATION                                        tion. In our case, the first spontaneous bleeding occurred
                                                                                  30 days after the third dose with negative investigations
                    I. Costanzo, M. Mazzetti, M.M. Poggi, G. Emmi,                for infectious, autoimmune, or neoplastic diseases. Even
                    E. Silvestri, C. Tamburini, D. Prisco                         though the association still remains purely temporal and
                                                                                  pathogenetic mechanisms have only been hypothesized,
                    Medicina Interna Interdisciplinare, AOU Careggi, Italy        the temporal association, the exclusion of other possible
                                                                                  trigger causes and the similar cases reported in the recent
                    Background: Acquired hemophilia A (AHA) is a rare             literature suggest a possible connection between this case
                    and potentially life-threatening bleeding disorder charac-    and Sars-CoV-2 mRNA-based vaccine.




                                                                                                          ly
                    terized by circulant autoantibodies directed against factor
                    VIII, resulting in an isolated prolongation of activated




                                                                                                   on
                    partial thromboplastin time (aPTT). Different conditions
                                                                                  PO074
                    have been associated to AHA such as neoplastic and
                    autoimmune diseases, pregnancy, infections, and drugs.




                                                                                             e
                    Between 2021 and 2022 13 reports of AHA following             ACQUIRED FACTOR XI INHIBITOR IN A CHILD
                    Sars-CoV-2 mRNA vaccine have been reported. We
                    report a case of AHA arose with skin hematomas and
                    macroscopic hematuria in a 74-year-old woman about a
                                                                                      us
                                                                                  WITH MEMBRANOUS GLOMERULONEPHRITIS

                                                                                  V. Nosratian, L. Banov, A. Naim, J. Svahn, A.C. Molinari
                    month after the third dose of mRNA Sars-CoV-2 vacci-
                                                                                  al
                    nation. Case Report: A 74-year-old woman presented to         Centro Emostasi e Trombosi, IRCCS G.Gaslini Genova,
                                                                          ci

                    our hospital for persistent hematuria onset on 10th           Italy
                    February 2022. She had received three mRNA Pfizer
                                                                    er


                    SarS-CoV-2 vaccines, the last one on 12th December            Background: Acquired deficiency of coagulation fac-
                    2021, followed by spontaneous hematomas and macro-            tors is rare, and is mainly associated with autoimmune
                                                              m



                    scopic hematuria respectively arose one and two months        disorders or malignancies. We describe a case of
                    after the vaccination. Laboratory tests showed normocyt-      acquired factor XI deficiency in a child affected by
                                                  om




                    ic anemia with Hb 10.5 g/L and isolated aPTT prolonga-        autoimmune disorder. Case Report: A 21 months old
                    tion (53.6 s) with normal PT (11,5 s) and normal platelets    male with autoimmune thyroiditis showed prolonged
                    count (215k). The coagulation factor assay showed an          aPTT (3,92 ratio). Not incubated mixing tests showed
                    isolated factor VIII deficiency with activity of 6.3% and     partial aPTT correction. During subsequent follow up
                                               -c




                    inhibitor titer of 2.5 BU, which was not corrected by the     he always showed persistent prolongation of aPTT no
                                                                                  more corrected by mixing test; second level tests at
                                       on




                    dilution test. AHA with low anti-factor VIII antibody titer
                    was diagnosed. Initially, she was treated with factor aVII    local institution revealed severe deficiency of factor XI
                    Eptacog alfa and prednisone 75 mg/die. After three days,      (<1%) and positive Lupus Anticoagulant test (dRVVT).
                                  N




                    due to persistent hematuria, she was treated with the new     No Factor XI gene mutation was found. His parents had
                    recombinant VIII porcine factor Susoctacog alfa.              normal Factor XI activity. Family history was unre-
                    Bleeding interruption was obtained with the restoration       markable and personal history did not reveal any bleed.
                    of hemoglobin levels. We excluded an anti-phospholipid        At the age of 53-months due to generalized edema,
                    antibodies’ syndrome, other autoimmune diseases and a         increased body weight and nephrotic proteinuria he
                    parainfectious origin. A paraneoplastic form was ruled        received oral prednisone 60 mg/m2/die. Proteinuria
                    out: neoplastic markers assay and occult stool’s blood        decreased, but its persistence the child received intra-
                    research, abdominal ultrasound, total body CT and PET,        venous (IV) metil-prednisolone boluses 10 mg/kg/die
                    mammography and breast ultrasound exams were nega-            for three days and prednisone was tapered. He was
                    tive. The abdominal CT showed clots in the right renal        referred to our Institution to undergo kidney biopsy
                    pelvis; a cysto-uretero-endoscopy was performed with          (KB). Two weeks from the steroid IV boluses a milder
                    negative result for neoplastic lesions. The immunophe-        prolongation of aPTT was observed (1,33 ratio) and the
                    notype showed a slight increase in CD8+ and CD4+ acti-        factor XI activity resulted increased (33.24%).
                    vated lymphocytes, in absence of significant quantitative     Acquired FXI deficiency secondary to autoantibodies
                    changes in the lymphocyte and leukocyte subpopula-            against FXI was supposed. Given the hemorrhagic risk
                    tions. We measured the anti S IgG too, positive with high     associated with the KB we anyway decided to give oral
                    titer (12,800), as expected in a recent SarS-CoV-2 vacci-     tranexamic acid and low dose rFVIIa before and three
                    nation, with negative IgM and RNA. Factor VIII levels         hours after KB that was safely performed. Membranous




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 130



         Poster




                    glomerulonephritis was diagnosed. Serial aPTT, FXI          Care, Internal Medicine and Medical Specialties
                    assays and FXI inhibitor were performed. While severe       (ProMISE), University of Palermo, Palermo, Italy;
                    Factor XI deficiency was persistently documented low        2Division of Internal Medicine, Hematology Service,

                    titer FXI inhibitor (1.4 Nijmegen Bethesda Units) was       Regional Hospital “ACardarelli”, Campobasso, Italy;
                    detected only once. Thromboelastometric global coagu-       3IRCCS Istituto Romagnolo per lo Studio dei Tumori
                    lation test (Rotem) showed normal clot activation time      (IRST)”Dino Amadori”, Meldola (FC), Italy
                    and strength. Thrombin generation assay will be per-
                    formed to better document the bleeding risk.                Background: Idiopathic thrombotic thrombocytopenic
                    Proteinuria recurred two months after IV steroid bolus-     purpura (iTTP) is an acquired thrombotic microan-
                    es and combined therapy was started with tacrolimus         giopathy caused by inhibition of the ADAMTS13 pro-
                    and, till now, two doses of rituximab. Coagulation          tein (a disintegrin and metalloproteinase with a throm-
                    parameters remained unchanged. The patient was there-       bospondin type 1 motif, member 13) by an inhibitory
                    fore sent to outpatient follow up initially every 2 weeks   antibody. iTTP is a rare disease and recurrences are
                    and then every 3 months (Table 1). Conclusions: The         even rarer. The etiopathogenesis of iTTP relapses is still
                    bleeding phenotype in FXI deficiency does not corre-        not completely clear and available evidences are scarce.
                    late with the FXI activity level, with evidence of bleed-   Infections, often viral, seem frequently triggering fac-
                    ing reported in heterozygotes with mild deficiency (FXI     tors     of    iTTP      relapses.     Both     laboratory
                    levels 20%–60%). Most of patients with severe defi-         (ADAMTS13<10%) and clinical recurrences of iTTP
                    ciency do not bleed spontaneously but can bleed seri-       have been associated with paucisymptomatic or compli-




                                                                                                        ly
                    ously during or after surgery. This lack of correlation     cated episodes of CMV and HSV mononucleosis, tuber-
                    between bleeding risk and FXI activity poses a signifi-     culosis, urinary tract infections and SARSCoV2 infec-




                                                                                                 on
                    cant therapeutic challenge. Acquired coagulation            tion. Case Report: We here report a series of 4 patients
                    inhibitors are autoantibodies that bind to coagulation      with a history of iTTP, followed up at our Institute, with
                    factors and neutralize their activity or accelerate their   a history of clinically significant infections during clin-




                                                                                           e
                    clearance. They include inhibitors against coagulation      ical remission of iTTP. Case 1: 33-yo man diagnosed
                    factors I, II, V, VII, VIII (acquired hemophilia A), IX
                    (acquired hemophilia B), X, XI, and XIII. Acquired
                    deficiency of FXI having been reported with autoim-
                                                                                    us
                                                                                with iTTP in June 2016, treated with steroid and PEX
                                                                                with the achievement of clinical and laboratory remis-
                                                                                sion. On April 201,7 the patient had laryngitis success-
                                                                                al
                    mune disorders (systemic lupus erythematosus,               fully treated with antibiotic therapy, ADAMTS13 activ-
                    rheumatoid arthritis, Crohn disease), or malignancies       ity in July 2017 was 102%. In February 2018 the patient
                                                                        ci

                    (chronic myelomonocytic or lymphocytic leukemias,           had pharyngitis successfully treated with antibiotic
                    and gastrointestinal adenocarcinoma) but only one case      therapy, ADAMTS13 dosage on March 2018 was
                                                                  er


                    of FXI inhibitor is reported in a child affected by mem-    100%. Case 2: 55-yo man diagnosed with iTTP in May
                    branoproliferative glomerulonephritis. Inhibitors           2020, treated with Caplacizumab and PEX with the
                                                             m



                    should be suspected when coagulation factor(s) defi-        achievement of partial clinical remission. In October
                    ciency is found in autoimmune diseases.                     2020, the patient had a subcutaneous abscess resulting
                                                 om




                                                                                from an infection of hernia mesh, for which he was
                    Tabella 1.                                                  treated with antibiotic therapy and vacuum-assisted clo-
                                                                                sure therapy (VAC) with success. During hospitaliza-
                                              -c




                                                                                tion, the patient had a clinical (Hb 9.2g/dL and PLT
                                                                                34000/mmc) and laboratory (ADAMTS13 1.3%)
                                      on




                                                                                relapse of iTTP disease, treated with Caplacizumab +
                                                                                PEX and Rituximab, with the achievement of hemato-
                                                                                logical remission. Case 3: 43-yo woman diagnosed with
                                  N




                                                                                iTTP in March 1998, treated with steroid, immunosup-
                                                                                pressants and FFP with complete hematological remis-
                                                                                sion. At follow-up in November 2021 and January 2022
                                                                                ADAMTS13 was 5.1% and 7.7% respectively, with
                                                                                normal haematochemical parameters. In February 2022
                                                                                the patient had pneumonia successfully treated with
                      PO075                                                     antibiotic therapy. During the hospital stay, the range of
                                                                                ADAMTS 13 values already detected before the infec-
                                                                                tion remained within the last control visit (ADAMTS13
                    DO INFECTIONS POSE A SIGNIFICANT RISK OF                    was found to be 7.7%, in March 2022 it was 5.7%, and
                    RELAPSE OF IDIOPATHIC THROMBOCYTOPENIC                      in April 2022 it was 6.8%), without any other detectable
                    PURPURA? A CASE SERIES                                      abnormality. Case 4: 52-yo woman diagnosed with
                                                                                iTTP in September 2019, treated with steroids,
                    C. Rotolo1, M. Mattana1, C. Cammarata1, R. Tomasello1,      Rituximab, FFP and PEX with the achievement of
                    G. Giordano2, A. Lucchesi3, S. Siragusa1,                   hematological remission. In September 2021 the patient
                    M. Napolitano1                                              had paucisymptomatic SARSCoV2 infection, resolved
                                                                                without specific therapy, in the course of which the
                    1Department   of Health Promotion, Mother and Child         blood count and ADAMTS13 levels remained normal.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 131



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Conclusions: Limited to the extreme rarity of the dis-         FVIII (233 B.U.) excluded the susoctocog alfa option.
                    ease, the real-life data collected in our case series do not   The patient had several recurrent hematomas on the
                    show a marked tendency for iTTP patients to relapse            shoulders and back, requiring packed red cells and
                    within the course of localized or systemic infections.         treatment with rFVIIa 90 ug/kg/ 6-8 hours. After 6
                    This is in contrast to the available literature, according     hours of wash out, Emicizumab 3mg/kg subcutaneously
                    to which activation of the immune system could trigger         weekly for 3 doses and then 1,5 mg/kg/weekly was
                    the production of autoantibodies. The pathogenetic             started. The day after the first Emicizumab dose the
                    mechanisms of ADAMTS13 inhibitor and iTTP relapse              aPTT was reduced by 60% (from aPTT 91 m“ to 26
                    need further investigation.                                    m“). The patient did not require additional rFVIIa infu-
                                                                                   sions, the hematomas reduced and on Day 6 since the
                    Table 1.                                                       Emicizumab had been started, the patient was dis-
                                                                                   charged (Table 1). The patient is actually coming in for
                                                                                   weekly follow up. Conclusions: Acquired haemophilia
                                                                                   A can be very challenging, even for experienced clini-
                                                                                   cians. Common bypassing agents as activated pro-
                                                                                   thrombin complex concentrates (APCC) and rFVIIa or
                                                                                   recombinant porcine FVIII concentrates in AHA bleed-
                                                                                   ing patients require very close monitoring and hospital-
                                                                                   ization. We describe a treatment for a refractory patient




                                                                                                          ly
                                                                                   that had been hospitalized for a long time and that had
                                                                                   been discharged after just two weekly Emicizumab 3




                                                                                                   on
                                                                                   mg/kg s.c. injections, with reduced hematomas, aPTT
                                                                                   in range and no bleeding.
                    PO076




                                                                                               e
                                                                                   Table 1.
                    EMICIZUMAB IN ACQUIRED HAEMOPHILIA A

                    F. Demartis, L. Pieri, R. Fjerza, S. Linari, G. Castaman
                                                                                       us
                                                                                   al
                    Sodc Malattie Emorragiche, Azienda Ospedaliero
                                                                           ci

                    Universitaria Careggi, Firenze, Italy
                                                                                   PO077
                                                                     er


                    Background: Acquired haemophilia A (AHA) is a rare
                    bleeding disorder caused by autoantibodies to FVIII            UNCONVENTIONAL USE OF RECOMBINANT
                                                               m



                    resulting in low FVIII. Bleeding is common, ranging            PORCINE FVIII IN ACQUIRED HAEMOPHILIA:
                    from 70 to 90% of cases at diagnosis. Activated pro-           THREE CASE REPORTS
                                                  om




                    thrombin complex concentrates(APCC) and recombi-
                    nant activated FVII (rFVIIa) are usually required to           C. Sella1, M. Bardetta1, C. Dainese1, F. Valeri1,
                    control bleeding, while plasma-derived or recombinant          A. Valpreda1,4, D. Grimaldi3, P. Martinelli1, D. Stella2,
                                               -c




                    FVIII concentrates, are effective only with low inhibitor      B. Bruno2, A. Borchiellini1
                    titers (<5 Bethesda Units). In recent years the use of
                                       on




                    recombinant porcine FVIII concentrates is an option for        1Regional   Centre for Haemorrhagic and Thrombotic
                    patients without cross-reactivity. Immunosuppressive           Diseases, Haematology Unit, City of Health and Science
                    therapy with corticosteroids (1 mg/Kg), cyclophos-             University Hospital of Molinette, Turin, Italy;
                                  N




                    phamide (2 mg/Kg) and /or rituximab (375 mg/m2), is            2Haematology Unit, City of Health and Science
                    used for inhibitor eradication, however the time               University Hospital of Molinette, Turin, Italy;
                    required to eradicate could be significantly long in           3Haematology Unit, Santa Croce e Carle Hospital,
                    patients with very high titer at presentation, thus putting    Cuneo, Italy; 4Central Laboratory Baldi and Riberi, City
                    the patient at risk of bleeding becuse of lack of effective    of Health and Science University Hospital of Molinette,
                    prophylaxis meanwhile. Emicizumab is a bispecific              Turin, Italy
                    antibody recognizing activated FIX and Factor X, with
                    FVIIIa-cofactor activity which allows effective prophy-        Background: Acquired haemophilia A (AHA) is due to
                    laxis in patients with congeniotal hemophilia A with           autoantibodies which interfere with the activity of clot-
                    and without inhibitors. A few cases of AHA treated with        ting factor VIII (FVIII). AHA is frequently caused by
                    this agent have been recently reported. Case Report: A         underlying conditions like systemic rheumatic disease,
                    66 year old male with a history of treatment-refractory        malignancy, drugs and postpartum. Treatment of AHA
                    AHA (four rituximab weekly infusions [375 mg/m2];              consists of tempestive control of bleeding and inhibitor
                    cortison [1 mg/kg/day]) and multiple bleedings, treated        eradication with immunosuppressive therapy (IST).
                    with rhFVIIa (90ug/Kg/4-6-h) came to our attention.            Hemostatic treatments include recombinant activated
                    The patient was diagnosed two months earlier with high         factor VII (rFVIIa), activated prothrombin complex con-
                    inhibitor titre 2480 B.U.; aPTT 96 m“ and no detectable        centrate (aPCC), both known as bypassing agents, and
                    FVIII. The cross-reactivity for recombinant porcine            susoctocog-alfa, a recombinant porcine FVIII (rpFVIII).




                                                                [Hematology Reports 2021; 13:s3]                                           [page 131]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 132



         Poster




                    The last one has been recently licensed for management         despite the presence of porcine FVIII inhibitors. Indeed
                    of acute bleeding in AHA. Unlike treatment with bypass-        the presence of rpFVIII inhibitors titre and the effect of
                    ing agents, rpFVIII can be monitored to provide a suc-         rpFVIII do not have correlation, as seen in previous stud-
                    cessful haemostatic effect. This advantage is important in     ies. Furthermore we describe use of rpFVIII in ambula-
                    patient with high thrombotic risk or those who need to         tory setting with reduced dosage (100 U/kg) every other
                    undergo surgery. Nevertheless, as shown by the litera-         day to prevent relapse. This schedule represents a
                    ture, only two case reports illustrate the use of rpFVIII in   favourable cost-efficacy alternative in management for
                    perioperative setting (ocular and abdominal surgery).          patients with high risk of re-bleeding (risk depends on
                    Instead, in patients with high haemorrhagic risk, after        site and extension of previous bleeding or concomitant
                    resolution of acute bleeding, is reasonable the usage of       conditions).
                    rpFVIII in prophylaxis schedule; only one case has been
                    described using prophylaxis dosage in outpatients. Case
                    Report: We report three cases of AHA in which was used         PO078
                    susoctocog-alfa in non-conventional schedule (Table 1).

                    Table 1.                                                       THALIDOMIDE AS EFFECTIVE DRUG FOR THE
                                                                                   CONTROL OF GATROINTESTINAL BLEEDINGS IN
                                                                                   CONGENITAL AND ACQUIRED BLEEDING
                                                                                   DISORDERS




                                                                                                           ly
                                                                                   A. Ferretti1, M. Basso1, C. Santoro3, D. Feliciani4,




                                                                                                   on
                                                                                   M.E. Riccioni4, S. Sorrentino1, E. De Candia1,2
                                                                                   1Malattie Emorragiche e Trombotiche, Fondazione




                                                                                              e
                                                                                   Policlinico Universitario Agostino Gemelli-IRCSS,
                                                                                   Roma, Italy; 2Dipartimento di Medicina e Chirurgia
                                                                                       us
                                                                                   Traslazionale, Università Cattolica del Sacro Cuore,
                                                                                   Roma, Italy; 3Ematologia, Azienda Ospedaliera
                                                                                   Universitaria Policlinico Umberto I, Roma, Italy;
                                                                                   al
                                                                                   4Gastroenterologia,      Fondazione       Policlinico
                                                                           ci

                                                                                   Universitario Agostino Gemelli-IRCSS, Roma, Italy
                                                                     er


                    Case 1: A 70-year-old woman with myasthenia gravis,            Background and Aims: Gastrointestinal bleeding
                    Hashimoto thyroiditis and glaucoma was hospitalized for        (GIB), both upper and lower, can be a well-recognised
                                                               m



                    right upper limb hematoma in AHA. Initially treated with       complications of congenital and acquired forms of von
                    steroid than cyclophosphamide. During hospitalization          Willebrand disease (vWD) and Glanzmann
                                                  om




                    she underwent surgical intervention for diverticular           Thrombasthenia (GT), generally arising in older
                    bowel perforation performed under rFVIIa coverage. In          patients and being a challenge for clinicians. The aim of
                    post-operative, for elevated rFVIIa dependency, was            this study is to describe the experience of GIB manage-
                                               -c




                    administered rpFVIII without haemorrhagic complica-            ment in patients with congenital and acquired coagula-
                    tions. Case 2: A 79-year-old man with history of stomach       tion defects and platelet alterations. Methods: Between
                                       on




                    carcinoma, suspected urothelial neoplasia and polymyal-        2016-2020, 5 patients affected by congenital and
                    gia rheumatica arrived at our attention for haematuria         acquired form of vWD and GT and severe GIB [3F/2M;
                    and diffuse hematomas. He was diagnosed with AHA               median age 66 years (66-77)] have been diagnosed and
                                  N




                    and treated with rFVIIa, subsequently changed with             treated successfully with thalidomide 50 or 100 mg/die.
                    rpFVIII for inadequate response. He underwent                  Diagnosis was made with clinical/laboratory features.
                    ureteroscopy with biopsy and laser ablation in the nearby      Data were collected from clinical charts. Results:
                    of neoplastic lesion under rpFVIII coverage without            Gastrointestinal bleeding is, potentially, a fatal condi-
                    complication. In the meanwhile, he received four lines of      tion due to loss of large volumes of blood.
                    IST for persistent higher titre of inhibitors. Case 3: A 31-   Angiodysplasia is thought to be one of the most com-
                    year-old woman, 60 days later partum, was admitted to          mon gastrointestinal vascular malformations associated
                    emergency room for anemia and right lower limb                 to GIB. Generally, the therapy is represented: 1) in con-
                    hematoma. She started therapy with rFVIIa and steroids         genital vWD by prophylaxis with VWF/factor VIII con-
                    and she obtained immediate response. Therefore, she was        centrates, 2) in congenital GT by platelet tranfusions or
                    dismissed eight days later. She was hospitalized again 5       rFVIIa infusions in patients with platelet alloantibodies,
                    days later for new bleeding, and she was treated with          3) in acquired form variability of response to standard
                    rFVIIa, than replaced with rpFVIII. The last one was           therapy is reported. In some cases, these therapies were
                    continued in ambulatory setting at reduced dosage. She         not always successful. Possible rescue therapies such as
                    had no relapse of AHA. Conclusions: We report a case           octreotide, thalidomide, lenalidomide, and tamoxifen
                    of successful peri-procedural haemostatic coverage with        were described in a few case reports. It is well known
                    rpFVIII in prevention of bleeding and another one after        the thalidomide effect on angiogenesis counteraction by
                    abdominal surgery (in immediate post-operative), both          its effects on vascular endothelial growth factor




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 133



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    (VEGF) and basic fibroblastic growth factor (bFGF)           correlation between SarsCov2 vaccines and AHA was
                    and, in few cases has been reported a positive effect in     supposed. Methods: Data about vaccines (type, dose
                    treatment of patients affected by vWDs and GT with           and timing) and AHA diagnosis and management were
                    GIB. Notably, we described 5 cases with vWD type 2           retrospectively collected. Patients’ features were sum-
                    (1), GT (1) and acquired vWD associated to MGUS (3)          marized in Table 1. Results: Our small study cohort
                    with recurrent severe GIB successfully treated, with dif-    consists of 5 females with median age at diagnosis of
                    ferent posology of thalidomide. In particular, two           77 years (range 34-80). Median FVIII activity and
                    patients with congenital disease exhibited GIB at older      median inhibitor titer at diagnosis was 0.019 IU/ml
                    age, caused by angiodysplasia, in absence of severe          (range 0.01-0.08) and 30 Bethesda Units/ml (range
                    bleeding history. After few months by starting thalido-      2.6-100), respectively. In 2 patients, relevant autoim-
                    mide 50 mg/die, acute and severe GIB were stopped.           mune comorbidities were found (Polymyalgia
                    Three patients with acquired VWD and concomitant             rheumatica and Scleroderma currently off-therapy).
                    IgG K MGUS had recurrent hospitalizations for acute          SarsCov2 vaccine was performed in all patients before
                    and severe melena caused by angiodysplasia. Two of           AHA diagnosis (2/5 after Vaxzevira® II dose, 2/5 after
                    them were primarily treated with intravenously               Cominarty® II dose and 1/5 after booster). No patients
                    immunoglobulins with good response even for a small          experienced SarsCov2 infection before AHA diagno-
                    period of time. After that, thalidomide administration       sis; SarsCov2 nasal swab turned positive after 2 weeks
                    was started resulting in increased hemoglobin levels.        of hospitalization in one patient. Median time from
                    Thalidomide treatment is still ongoing in all the patients   vaccine to AHA diagnosis was 121 days (range 74-




                                                                                                        ly
                    with an improvement in the quality of life, obtaining a      222); 2/6 and 1/6 patients had cutaneous and muscular-
                    striking and stable reduction of GIB and of the require-     cutaneous bleeds at diagnosis respectively. Median




                                                                                                 on
                    ment of platelet and blood transfusions. Conclusions:        time from vaccine to diagnosis and from vaccine to
                    Although the exact mechanism by which thalidomide            clinical onset was 97 days and 56 days, respectively.
                    exerts antiangiogenic effects is not fully understood, we    Two Patients had no bleeds and AHA has been diag-




                                                                                            e
                    can describe its successfully use in this particular con-    nosed after routine blood exams. In these cases, AHA
                    test of patients with severe GIB. This preliminary eval-
                    uation needs to be carried out on a larger population to
                    better understand both good and side effects of thalido-
                                                                                     us
                                                                                 onset remains unknown and time from vaccine to diag-
                                                                                 nosis was longer (134 and 121 days). All patients failed
                                                                                 first line immunosuppressor therapy (steroid, pred-
                                                                                 al
                    mide in patients with vWD and GT with GIB consider-          nisone 1mg/kg/day) so a second line immunosuppres-
                    ing the possibility to standardize this treatment.           sor therapy (Rituximab® 375mg/sq weekly, 4 weeks)
                                                                         ci

                                                                                 was required with 5 complete responses; in particular,
                                                                                 3/5 patients didn’t achieve remission during steroids
                                                                   er


                    PO079                                                        and 2 patients relapsed after steroid discontinuation.
                                                                                 Recombinant porcine FVIII concentrate (Obizur®)
                                                             m



                                                                                 was used in 3/5 patients with very good bleed control.
                    INCREASED INCIDENCE OF ACQUIRED
                                                                                 Median follow-up is 5.8 months (range 3.8-6.4).
                                                 om




                    HAEMOPHILIA A DURING THE SARSCOV2                            Conclusions: Correlation between SarsCov2 vaccine
                    VACCINE CAMPAIGN: PATHOPHYSIOLOGICAL                         and AHA onset can not be demonstrated by our small
                    RELATIONSHIP OR INCREASED DIAGNOSIS                          sample size. However, without other triggers it is rea-
                                              -c




                    VIGILANCE?                                                   sonable to assume a possible role of sarscov2 immu-
                                                                                 nization in AHA pathophysiological process.
                                       on




                    J. Agnelli Giacchello, P. Sciancalepore, L. Contino,         Furthermore, a delayed diagnosis of AHA it is common
                    R. Santi                                                     due to low incidence, inadequate clinician awareness
                                                                                 and to large variety of the bleeding symptoms (asymp-
                                  N




                    SSD Emostasi e Trombosi, Ospedale S.SAntonio e Biagio        tomatic case vs life-threating bleeds). This trend could
                    e CArrigo, Alessandria, Italy                                explain the wide range in time from vaccine to diagno-
                                                                                 sis in our population. Efficacy and safety of Obizur®
                    Background and Aims: Acquired Hemophilia A                   and Rituximab® was confirmed.
                    (AHA) is a rare autoimmune disorder caused by
                    autoantibodies affecting clotting factor VIII activity       Table 1.
                    leading to a bleeding disorder. Population-based unse-
                    lected cohorts have calculated an incidence of acquired
                    factor VIII inhibitor of 1.3 to 1.5 cases per million pop-
                    ulation per year. An increased incidence of AHA from
                    October 2021 to March 2022 was observed in
                    Alessandria area: 5 new diagnoses in a population of
                    about 400.000 residents (approximately 20 times more
                    than expected). Previously, several reports of possible
                    correlation between vaccines and immune disorders
                    onset have been reported. In recent few months at least
                    10 cases of AHA post SarCcov2 vaccine were reported.
                    Considering the wide vaccine campaign, a possible




                                                              [Hematology Reports 2021; 13:s3]                                          [page 133]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 134



         Poster




                      PO080                                                     allow the patient to safely undergo surgery, avoiding or
                                                                                reducing hemorrhagic manifestations.
                                                                                We consider useful to evaluate the vWF-RCo/vWF-Ag
                    ACQUIRED TYPE 2 WILLEBRAND’S DISEASE IN                     ratio in all patients with TE, to limit the risks of anti-
                    PATIENTS WITH ESSENTIAL THROMBOCYTHEMIA                     thrombotic prophylaxis with aspirin as much as possi-
                                                                                ble. Finally, literature reports a higher incidence of
                    D. Dragonetti1, M. Becce1, S.P. Pascale2, S. Coluzzi2,      Willebrand type 2 in patients with the JAK2 mutation,
                    R. Nuccorini2, M. Giuseffi1, A.M. Calabrese1,               but both our patients have the Calreticulin type 2 muta-
                    A. Picerno1, A. Matturro2, A. Amendola2, I. Chitarrelli2,   tion, so we will further investigate all our TE patients to
                    R. Iula2, K. Ferrara2, D. Caputo2, M. Cimminiello2,         see if there may be a correlation with this type of muta-
                    N. Filardi2, D. Vertone2, M. Pizzuti2                       tion as well.
                    1Department of Clinical Laboratory; 2Department of          Table 1.
                    Hematology; Regional Hospital “San Carlo”, Potenza,
                    Italy

                    Background and Aims: Patients with Essential
                    Thrombocythemia (TE) can have both thrombotic and
                    hemorrhagic events. In a study of 809 patients, 65%




                                                                                                        ly
                    experienced thrombosis, 15% both thrombosis and
                    bleeding, and 1.4% only bleeding, that is often gas-




                                                                                                 on
                    trointestinal and muco-cutaneous, as occurs in middle-      PO081
                    severe Willebrand’s disease, and is promoted by sur-
                    gery. It is more common with marked thrombocytosis,         A CASE OF CONGENITAL GLANZMANN




                                                                                           e
                    but can also occur with platelets slightly above            THROMBASTHENIA WITH A NEW MUTATION IN
                    400,000. One of the causes of bleeding is a reduction in
                    vWF-RCo due to a decrease in high molecular weight
                    multimers, with normal antigenic concentration and
                                                                                    us
                                                                                ITGA2B GENE AND EFFECTIVE CONTROL OF
                                                                                GASTROINTESTINAL BLEEDING BY THALIDOMIDE
                    reduced vWF-RCo/vWF-Ag ratio. In these cases
                                                                                al
                                                                                S. Sorrentino1, D. Feliciani2, A. Ferretti1, A. Minucci3,
                    Desmopressin (DDAVP) can induce a dose-dependent            E. De Paolis3, P. Concolino3, E. De Candia1,4
                                                                        ci

                    increase in high molecular weight multimers of vWF
                    with improvement in vWF: Ag/vWF:RCo. After few              1Unità Operativa Malattie Emorragiche e Trombotiche,
                                                                  er


                    hours, there is clearance of multimers, whose rapidity      Area di Ematologia, Fondazione Policlinico
                    is correlated with platelet count. so it may be necessary   Universitario “Agostino Gemelli”, IRCCS, Rome, Italy;
                                                             m



                    to readminister DDAVP.We report the case of two             2UOC di Medicina Interna e Gastroenterologia,
                    patients, with TE, both with Calreticulin type ins 5        Dipartimento di Scienze Mediche e Chirurgiche,
                                                 om




                    mutation, not on cytoreductive therapy. The first patient   Fondazione Policlinico Universitario A. Gemelli IRCCS,
                    needed laparoscopic multiple myomectomy, the second         Rome, Italy; 3Molecular and Genomic Diagnostics Unit,
                    had bleeding after dental extraction. In both of them on    Fondazione Policlinico Universitario AGemelli IRCCS,
                    screening examinations there was a reduction of
                                              -c




                                                                                Rome, Italy; 4Dipartimento di Medicina e Chirurgia
                    vWF:RCo, with normal value of vWF:Ag, so we per-            Traslazionale, Università Cattolica Del Sacro Cuore,
                                      on




                    formed a test with DDAVP 0.3 µg/kg with improve-            Rome, Italy
                    ment of vWF:RCo. Methods: For the quantitative
                    determination of vWF: Ag and of vWF-RCo, we used            Background: Glanzmann Thrombasthenia (GT) is an
                                  N




                    two automated latex enhanced immunoassay in human           autosomal recessive disorder of platelet function
                    citrated plasma on IL Coagulation Systems. Results:         caused by quantitative or qualitative defects in inte-
                    The results obtained from the laboratory tests (Table 1),   grins αIIb and β3. The most common symptoms of GT
                    for the two patients showed a remarkable increase in        are spontaneous mucocutanous bleedings, unremitting
                    vWF-Ag concentrations, and in its activity, with an         bleeding after injury or surgery, menorrhagia in
                    increase in the ratio vWF-RCo / vWF: Ag and improve-        women. Spontaneous upper gastrointestinal bleeding
                    ment in PFA. The first patient underwent surgery after      (GIB), causing recurrent episodes of severe anemia
                    administration of DDAVP every 12 hours for 3 days,          and requiring hospitalization for invasive procedures
                    without hemorrhagic complications. The second               for bleeding control and red blood cells transfusions,
                    patient, is waiting for a new dental surgery.               may also occurr. Angiodysplasias sustaining GIB have
                    Conclusions: We confirm the evidence in published lit-      been described in GT patients. There is no specific
                    erature on the possibility of hemostatic alterations in     therapy to control GIBs, except for local treatments,
                    patients with TE due to the reduction of vWF-RCo (and       and systemic hemostatic therapy such as platelet trans-
                    that this phenomenon is possible even when platelet         fusions, antifibrinolytitcs, rFVIIa. We describe a case
                    values are less than one million) and underline that the    of GT with a new mutation in the ITGA2B gene who
                    administration of DDAVP can re-establish a proper,          showed a good GIB control under therapy with the
                    although transient, balance between vWF-RCo/vWF-            antiangionic drug thalidomide. Case Report: A 62-
                    Ag and optimize hemostatic mechanisms. This will            year-old woman was hospitalized in our hospital for




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 135



                                                                       XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    rectal bleeding and melena sustained by multiple gas-      PO082
                    trointestinal angiodysplasia (GIA) and active bleeding
                    at caecum. She reported history of spontaneous muco-
                    cutaneous hemorrhagic manifestations, nosebleeds,          PROPHYLAXIS WITH PDVWF IN A OBESE
                    gingivitis, frequent cutaneous bleeding, anemia since      WOMAN AFFECTED BY VWD AND MUTIFOCAL
                    childhood. The patient reported that an undefined          ANGIODYLASIA: A CHALLENGING SCENARIO
                    platelet dysfunction was diagnosed and platelet trans-
                    fusions were required during some bleeding episodes        R. Tomasello1, M. Mattana1, C. Rotolo1, C. Cammarata1,
                    in other hospitals. She also reported history of sys-      A. Lucchesi2, G. Giordano3, S. Siragusa1,
                    temic lupus erythematosus and fluctuating thrombocy-       M. Napolitano1
                    topenia responsive to corticosteroids in the last years.
                                                                               1Department  of Health Promotion, Mother and Child
                    Platelet function studies during hospitalization in our
                    institution showed severe reduction of platelet aggre-     Care, Internal Medicine and Medical Specialties
                    gation to all stimuli except to ristocetin together with   (ProMISE), University of Palermo, Palermo, Italy;
                                                                               2IRCCS Istituto Romagnolo per lo Studio dei Tumori
                    strong reduction of glicoproteins αIIb and IIIb expres-
                    sion (Figure 1) and TG was diagnosed. The genetic          (IRST) “Dino Amadori”, Meldola (FC), Italy; 3Division
                    test demonstrated a new undescribed homozygotic            of Internal Medicine, Hematology Service, Regional
                    mutation in the ITGA2B gene [c.1096C>T and                 Hospital “ACardarelli”, Campobasso, Italy
                    p.(Arg366*)]. Actively bleeding angiodysplasia was




                                                                                                      ly
                    demonstrated and treated by argon plasma coagulation       Background: von Willebrand disease (VWD) is the
                    on two different episodes at a distance of one month.      most common inherited coagulopathy worldiwide.




                                                                                               on
                    Due to progressive increase of GIB episodes requiring      VWD has a strong association with congenital
                    hospitalization in the last two years, the patient was     angiodysplasias of the ileum, vascular malformations
                    treated off-label with thalidomide 50 mg/day. The          with a tendency to cause intestinal haemorrhages result-




                                                                                         e
                    patient was stable for 6 months, after which a new         ing in melena. More than 90% of patients with ileal
                    hospitalization for GIA of cecal fundus was necessary.
                    The thalidomide posology was increased to 100
                    mg/day, but it was reduced to 50 mg/day after two
                                                                                  us
                                                                               angiodysplasia are eventually diagnosed with VWD.
                                                                               The management of a haemorrhagic coagulopathy asso-
                                                                               ciated with vascular malformations poses several chal-
                    months because of side effects such as drowsiness,         lenges to clinicians. Case Report: A 60-yo obese
                                                                               al
                    mental confusion and constipation. After 12 months of      patient with a known diagnosis of VWD type 1, in the
                                                                       ci

                    treatment, she is in good general clinical conditions,     past medical history treated on demand with plasma-
                    hemoglobin is stable and no new GIBs occurred.             derived coagulation factor VIII-rich Von Willebrand
                                                                 er


                    Conclusions: GIB is a recurrent symptom in some GT         factor (pdVWF/FVIII) for mucosal bleedings in
                    patients, requiring frequent hospitalizations and          September 2021 was admitted to the E.R. for diarrhoea
                                                            m



                    urgent medical therapy, thus severely affecting their      with melena and severe anemia. She was transfused
                    quality of life. Thalidomide is an antiangiogenic drug     with 2 pRBC and discharged. Due to persistence of
                                                om




                    that has been used for the control of GIB in patients      melena, she was admitted to the Department of Internal
                    with congenital and acquired vonWillebrand diseases        Medicine and started replacement therapy with
                    and in patients with hereditary hemorrhagic teleangec-     pdVWF/FVIII. During her hospitalization, following
                    tasia. One case of GT patient treated with thalidomide
                                             -c




                                                                               the onset of bilateral pain and oedema of the upper
                    has previously been described, resulting in a signifi-     limbs, superficial vein thrombosis (SVT) was diag-
                    cant decrease of blood transfusion, bleeding episodes
                                      on




                                                                               nosed by Ultrasound, FVIII:c levels resulted 195%. The
                    and hospitalizations (ref). We found a new homozy-         patient thus needed anticoagulant treatment with Low
                    gotic mutation in the ITGA2B gene. In addition, we         Molecular Weight Heparin (LMWH). During the hospi-
                                 N




                    confirm that gastrointestinal angiodysplasias may sus-     tal stay endoscopic evaluation of the gastrointestinal
                    tain recurrent GIB in GT patients and that thalidomide     tract were performed, videocapsular endoscopy
                    can be effective for the control of GIB in these           revealed multiple angiodysplasias at medial and distal
                    patients, although possible adverse effects may limit      ileal level. She was discharged after two weeks with
                    its use.                                                   resolution of melena. Concurrently, for the indication to
                                                                               LMWH treatment of SVT, prophylaxis with
                                                                               pdVWF/FVIII was administered. In November 2021,
                                                                               due to the recurrence of melena, the patient was admit-
                                                                               ted to the Gastroenterology unit where, due to severe
                                                                               sideropenic anemia, she was treated with pRBC and
                                                                               intravenous iron carboxymaltose, subcutaneous
                                                                               octeotride and pdVWF/FVIII were administered to con-
                                                                               trol bleeding. Surgical treatment of angiodypslasia was
                                                                               deemed too risky due to severe obesity. At discharge, a
                                                                               purified plasma derived vWF(pdvWF) concentrate pro-
                                                                               phylaxis was chosen to counterbalance the high throm-
                                                                               botic risk. Prophylaxis was effective and safe. In
                    Figure 1.                                                  February 2022, the patient was however again admitted




                                                             [Hematology Reports 2021; 13:s3]                                          [page 135]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 136



         Poster




                    to the E.R. for melena with severe anemia. Endoscopic          patients. A small number of patients with congenital
                    treatment of angiodysplasia was than planned. On April         fibrinogen deficiencies (CFD) are evaluated with these
                    2022, the endoscopic procedure was perfomed under              tools without definitive results. Aims: To compare the
                    treatment with pdvWF with a partial removal of the             results of the ISTH-BAT with the EN-RBD-BSS in
                    tumors without complications. Prophylaxis was then             patients with CFD. We further evaluated the correlation
                    tapered on FVIII:c, vWF:Ag and vWF:Ricof levels, in            between bleeding scores (BS) and fibrinogen levels.
                    absence of bleeding. Currently then patient receives           Methods: In total, 100 (55% females) Iranian patients
                    pdvWF twice weekly, mantaining stable haemoglobin              with CFD were included. Routine coagulation and fib-
                    levels, in absence of melena (Figure 1). Conclusions:          rinogen-specific tests including antigen (Fg:Ag) and its
                    The association between VWD and ileal angiodysplasia           functional activity (Fg:C) were performed. Patients
                    is not infrequent and difficult to manage, it carries a        were classified using Fg:C/Fg:Ag ratio and they were
                    high risk of severe anemia due to martial deficiency           divided into four categories of bleeding severity
                    caused by chronic haemorrhages. In the treatment of            (asymptomatic, grade I, grade II, grade III). The ISTH-
                    gastrointestinal bleeding in this patient with high            BAT and the EN-RBD-BSS were used to assess the BS
                    thrombosis risk, exposure to high doses of                     of all patients. Pearson’s correlation was performed
                    pdVWF/FVIII was associated with SVT and required a             using IBM-SPSS Statistics 26. Results: The mean age
                    difficult combination with LMWH, while prolonged               of patients were 18.5 (SD±15.9) years. The median
                    prophylaxis with pdVWF resulted safe and equally               (range) of the ISTH-BAT and EN-RBD-BSS were 4 (0-
                    effective in controlling bleeding. This finding may be         16) and 2.21 (-1.49-6.71), respectively. In general, a




                                                                                                          ly
                    related to the difference in FVIII:c levels reached dur-       moderate positive correlation (r=0.597, P<0.001) was
                    ing therapy with pdWVF/FVIII. Prophylaxis in these




                                                                                                   on
                                                                                   found between the ISTH-BAT score and EN-RBD-
                    patients is not independent from the specific therapy of       BSS. The correlation between Fg:Ag and the ISTH-
                    the vascular lesions, which is crucial to control bleed-       BAT scores was weak negative (r=-0.407, P<0.001),
                    ing. In the above reported case, a partial removal of the      comparable to the correlation between Fg:C and ISTH-




                                                                                             e
                    tumors led to the resolution of bleeding.                      BAT scores (r=-0.423, P<0.001). There was a weak


                                                                                   BSS
                                                                                      us
                                                                                   negative correlation between Fg:Ag and EN-RBD-BSS
                                                                                   (r=-0.466, P<0.001), and between Fg:C and EN-RBD-
                                                                                                scores         (r=-0.446,        P<0.001).
                                                                                   al
                                                                                   Hypodysfibrinogenemic patients had the strongest cor-
                                                                                   relation between the EN-RBD-BSS and ISTH-BAT (r=-
                                                                            ci

                                                                                   0.916, P=0.001). Excluding dysfibrinogenemic
                                                                                   patients, there was a significant association between
                                                                        er


                                                                                   bleeding severity and Fg:C (r=-0.402, P<0.001), and
                                                                                   Fg:Ag (r=-0.316, P=0.005). Conclusions: Based on the
                                                                  m



                                                                                   good correlation between the two systems, it appears
                                                                                   that in addition to the ISTH-BAT score, the EN-RBD-
                                                    om




                    Figure 1. FVIII:c according to therapy.
                                                                                   BSS could also be useful in CFD patients. The BS
                                                                                   showed a significant negative correlation with fibrino-
                                                                                   gen antigen/activity levels.
                                                 -c




                      PO083
                                         on




                                                                                   PO084
                    THE UTILITY OF BLEEDING ASSESSMENT TOOLS
                    IN FIBRINOGEN DEFICIENCIES
                                   N




                                                                                   GENDER DIFFERENCES IN THROMBOTIC AND
                    S. Mohsenian1, O.    Seidizadeh2,3,
                                                   R.       M.Palla3,   Jazebi4,   HEMORRHAGIC ISSUES AFTER COVID-19
                    A. Azarkeivan1, S. Moazezi4, M.R. Baghaeipour4,                VACCINATION
                    M. Menegatti2, F. Peyvandi2,3
                                                                                   E. Grandone1,2, S. Chiocca, S. Castelvecchio, R. Nappi
                    1Blood  Transfusion Research Center, High Institute for        on behalf of representatives for Gender Medicine of
                    Research and Education in Transfusion Medicine,                Scientific Hospitalization and Treatment Institutes–
                    Tehran, Iran; 2Fondazione IRCCS Ca’Granda Ospedale             Italian Ministry of Health
                    Maggiore Policlinico, Angelo Bianchi Bonomi
                                                                                   1Thrombosis and Haemostasis Unit, IRCCS “Casa
                    Hemophilia and Thrombosis Center and Fondazione
                    Luigi Villa, Milan, Italy; 3Department of                      Sollievo della Sofferenza”, San Giovanni Rotondo (FG);
                                                                                   2Ob/Gyn Department, University of Foggia, Foggia;
                    Pathophysiology and Transplantation, Università degli
                                                                                   3IEO, Istituto Europeo di Oncologia IRCCS, Milano;
                    Studi di Milano, Milan, Italy; 4Iranian Comprehensive
                                                                                   4IRCCS Policlinico San Donato, San Donato Milanese
                    Hemophilia Care Center, Tehran, Iran
                                                                                   (MI); 5IRCCS Istituto Ortopedico Rizzoli, Bologna;
                                                                                   6IRCCS Fondazione Policlinico S. Matteo, Pavia, Italy
                    Background: Bleeding-assessment-tools (BAT) or
                    bleeding-score-systems (BSS) are developed to address
                    the challenges associated with the evaluation of               Gender medicine deals with differences in approaching




         [page 136]                                           [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 137



                                                                           XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    diagnostic work-up and management according to gen-            ting in different waves of the pandemic. Methods:
                    der. Although the issue is relevant in every field of med-     Multicenter, prospective study of patients with COVID-
                    icine, it is often neglected. The recent pandemic due to       19 pneumonia admitted to Internal Medicine units. All
                    SARS-CoV-2 virus raises this matter even more urgent-          patients were screened for DVT of the lower limbs with
                    ly. In fact, available literature has suggested a higher       serial CUS. The primary end-point of the study was to
                    number of deaths among infected males than in females          assess the incidence of DVT diagnosed by CUS in the
                    and more side effects in women than in men recipients          different waves of SARS-CoV-2 pandemic. Results:
                    of certain vaccines against COVID-19. In this narrative        Two periods of time, named first wave and subsequent
                    review, we critically revised sex-disaggregated data on        waves were considered, and a total of 363 consecutive
                    thrombotic and bleeding events associated with vaccina-        patients with moderate-severe COVID-19 pneumonia
                    tion against COVID-19. Thrombotic complications are            were enrolled. The pooled incidence of DVT in the two
                    by far more frequently reported than bleeding events           cohorts was 8%; incidence of DVT was 13.5% (6.8%
                    after vaccination and are mostly observed in young             proximal, 6.7% distal) in the first wave, and 4.2%
                    women who are administered with viral-vectored vac-            (2.3% proximal, 1.9% distal) in the subsequent waves
                    cines. Bleeding complications are mainly reported as           (p=0.002). Patients enrolled in the first and the second
                    aggregated data, whereas thrombocytopenia is reported          wave had similar clinical characteristics, and thrombot-
                    to occur in the presence or absence of thrombotic com-         ic risk profile as assessed by PADUA score (mean score
                    plications. This information is relevant, as it underlines     6.14 vs 6.31; p=0.155); a significant difference in anti-
                    the need to differentiate thrombocytopenia occurring           coagulant regimen and initiation of thromboprophylax-




                                                                                                            ly
                    with- or without -thrombosis; indeed, management and           is at home (8.1% vs 25.1%; p<0.001) was observed.
                    prognosis are different according to the presence of




                                                                                                    on
                                                                                   Conclusions: The incidence of DVT, diagnosed
                    thrombotic events. Conflicting data are available on the       through a surveillance protocol by serial CUS of the
                    prevalence of bleeding events according to sex in recip-       lower limbs, in acutely ill patients with COVID-19
                    ients of viral-vectored vaccines. Lastly, clinical data on     pneumonia hospitalized outside the ICU showed a sig-




                                                                                               e
                    vaccination against SARS-CoV-2 during pregnancy                nificant reduction over time during the different waves
                    show that its safety-efficacy profile is high. However,
                    further data are needed and will be obtained from ongo-
                    ing trials. In conclusion, available literature on sex -dis-
                                                                                       us
                                                                                   of the pandemic.
                                                                                   al
                    aggregated data and on the possible role of hormones in        PO086
                    thrombotic and bleeding complications after vaccination
                                                                           ci

                    against COVID-19 is still largely lacking. In view of the
                                                                                   LABORATORY EVALUATION OF POST-COVID19
                    role played by platelets in the pathogenesis mechanism
                                                                     er


                    of both thrombotic and bleeding complications, future          PATIENTS WITH ACUTE VENOUS
                    studies should be focused on platelets and on global tests     THROMBOEMBOLISM DURING
                                                               m



                    as thrombin generation assays.                                 HOSPITALIZATION: THE BERGAMO EXPERIENCE
                                                  om




                                                                                   C. Ambaglio1, S.V. Benatti2, C.J. Tartari1,
                    PO085                                                          C. Giaccherini1, L. Russo1, M. Marchetti1,
                                                                                   A.M. Palladino3, F. Schieppati1, S. Venturelli2,
                                                                                   L. Barcella1, M. Rizzi2, A. Falanga1,3
                                               -c




                    TRENDS IN THE INCIDENCE OF LOWER LIMB
                    DEEP VEIN THROMBOSIS IN PATIENTS WITH                          1Division
                                       on




                                                                                              of Immunohematology and Transfusion
                    COVID-19 PNEUMONIA DURING THE EARLIER                          Medicine, Papa Giovanni XXIII Hospital, Bergamo;
                    AND LATER WAVES OF SARS-COV-2 PANDEMIC:                        2Division of Infectious Diseases, Papa Giovanni XXIII
                                  N




                    A MULTICENTER PROSPECTIVE STUDY                                Hospital, Bergamo; 3Medicine and Surgery, University
                                                                                   of Milan-Bicocca, Milan, Italy
                    L. Corbo1, F. Mani1, R. Marcucci2, G. Guazzini1,
                    L. Lastraioli1, F. Luise1, L. Maddaluni1, A. Milia1,           Background and Aims: In a cohort of COVID-19 sur-
                    L. Sammicheli1, A. Fortini3, F. Pieralli1                      vivors who experienced venous thromboembolism
                                                                                   (VTE) during the acute phase and followed up at out out-
                    1COVID-19 Intermediate    Care Unit, Careggi University        patient clinic, we evaluated whether alterations in coagu-
                    Hospital, Florence, Italy; 2Department of Clinical and         lation and inflammatory parameters were persisting after
                    Experimental Medicine, University of Florence and              hospital discharge. Methods: An outpatient service
                    Careggi Hospital, Florence, Italy; 3Internal Medicine          specifically dedicated to follow up COVID-19 survivors
                    COVID-19 Unit, San Giovanni di Dio Hospital,                   was set up at our institution. A retrospective analysis of a
                    Florence, Italy                                                cohort of 1,563 patients enrolled from May 2020 to
                                                                                   November 2020 was performed. Data were obtained on
                    Background and Aims: The aim of this study was to              the occurrence or not of acute COVID-19 associated
                    evaluate the trend of the incidence of deep vein throm-        VTE, the site of thrombosis, ongoing anticoagulant treat-
                    bosis (DVT) of the lower limbs, using serial compres-          ment and the results of laboratory tests performed at the
                    sion ultrasound (CUS) surveillance, in patients hospi-         first post-discharge assessment visit, including blood cell
                    talized with COVID-19 pneumonia in a non-ICU set-              count, D-dimer, fibrinogen and CRP. Statistical analysis




                                                                [Hematology Reports 2021; 13:s3]                                              [page 137]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 138



         Poster




                    was performed using SPSS v21 software. Results of lab-     markers sCD146 (Biocytex, France), s-thrombomod-
                    oratory tests were expressed as median values (5th-95th    ulin (sTM) and se-selectin (R&D Systems, MN, US) in
                    percentile). Results: Fifty-six out of 1,563 COVID-19      190 samples from 101 patients with laboratory proven
                    survivors [43M/13F, median age 62 years (40-80.5)          coronavirus 2 (SARS-CoV-2) infection accessing the
                    3.58%] experienced VTE during the acute phase as fol-      ER of the San Raffaele Hospital in March 2020. Control
                    lows: 9 isolated proximal DVT (4 in upper extremity), 45   samples were from 45 apparently healthy volunteers.
                    isolated PE (7 subsegmental PE, 27 segmental PE, 11        Eighteen patients were not hospitalized (NH); 46
                    central pulmonary artery thrombosis), 2 DVT + PE, with     patients were admitted to medical wards (MW), and 37
                    subsegmental and segmental PE representing 60.7% of        to intensive care units (ICU). Samples from 25 ICU
                    all VTE. The first outpatient evaluation was performed     patients (9 survivors and 16 non-survivors) were addi-
                    between 41 and 222 days post-admission (average 181).      tionally tested after a median interval of 7 days (T2).
                    Treatment of VTE consisted in LMWH (n=11), vitamin         Samples from 40 MW patients and 19 ICU patients (all
                    K antagonist (n=2) and DOAC (n=43). Only two patients      survivors) were tested at a median interval of 3 months
                    had already stopped anticoagulant therapy at the time of   (T3). The hospital’s EC approved the study, registered
                    laboratory evaluation and presented normal values of D-    on ClinicalTrials.gov (NCT04318366). Results: At
                    Dimer. Laboratory data analyses revealed that 13/56        baseline DD, TFPI, and sTM were higher and sCD146
                    patients had persistent D-dimer >500 ng/ml, 11/56 had      lower in patients than in controls (p<0.0001). Trends
                    D-dimer >age-adjusted cut-off. 23,6% of VTE subjects       according to the severity of infection were only
                    had increased levels of both D-Dimer (>500 ng/ml) and      observed for DD and sCD146 (Table 1). T2 samples




                                                                                                      ly
                    fibrinogen (>450 mg/dl), and 11% had elevated levels of    showed increased levels of sTM, se-selectin, and
                                                                               decreased levels of PC and PS (p≤0.01), with no signif-




                                                                                               on
                    CRP (>1 mg/dl), while no abnormalities were observed
                    in the platelet and leucocyte counts. CRP significantly    icant difference between survivors and non-survivors.
                    correlated with fibrinogen and D-Dimer levels (r=0.791,    T3 samples showed increased PC and PS and reduced
                    p<0.0001). No patients had VTE recurrence or bleeding      DD and TFPI (p<0.0001) compared to T1 samples.




                                                                                          e
                    complications at the time of first follow up visit.        Conclusions: Compared to changes in DD levels,
                    Conclusions: Anticoagulant treatment of COVID-19
                    VTE seems to be clinically safe and effective, nonethe-
                    less laboratory analyses suggest a persistence of pro-
                                                                                   us
                                                                               changes in endothelial markers and natural anticoagu-
                                                                               lants are relatively moderate in covid-19 patients. The
                                                                               later increase in PS and PC antigen levels suggests a
                                                                               rebound phenomenon.
                                                                               al
                    coagulant and inflammatory alterations in spite of ongo-
                    ing anticoagulants. Further observations are needed to
                                                                       ci

                                                                               Table 1.
                    establish on long-term persistence of these laboratory
                    features and long-term risk of VTE recurrence even after
                                                                 er


                    the end of therapy.
                                                            m




                      PO087
                                                om




                    NATURAL ANTICOAGULANTS AND ENDOTHELIAL
                    MARKERS IN COVID-19 PATIENTS:                              PO088
                                             -c




                    A LONGITUDINAL EVALUATION
                                                                               THROMBOTIC THROMBOCYTOPENIA FOLLOWING
                                      on




                    P. Della Valle, C. Tresoldi, N. Maugeri, L. Apruzzi,       SYSTEMIC INJECTION OF THE RECOMBINANT
                    S. Calvisi, M. Ripa, G.A. Ramirez, P. Rovere-Querini,      ADENOVIRAL VECTOR SIMIAN ADENOVIRUS Y25
                                 N




                    G. Landoni, A. Castagna, M. Tresoldi, G. Martino,          SEROTYPE (CHADY25): REPORT OF TWO CASES
                    F. Ciceri, A. Manfredi, A. D’Angelo, on behalf of          AND A PERSPECTIVE ON PATHOGENESIS
                    COVID-BioB Study Group
                                                                               C. Cammarata1, M. Mattana1, C. Rotolo1,
                    Scientific Institute Ospedale San Raffaele and Vita e      R. Tomasello1, A. Lucchesi2, G. Giordano3, S. Siragusa1,
                    Salute University, Milano, Italy                           M. Napolitano1

                    Background and Aims: A combination of cytokines,           1Department of Health Promotion, Mother and Child
                    activated complement, platelets, endothelial cells and     Care, Internal Medicine and Medical Specialties
                    microvescicles appear to sustain the coagulopathy in       (ProMISE), University of Palermo, Palermo, Italy;
                    severe covid-19 infection. We evaluated changes in nat-    2IRCCS Istituto Romagnolo per lo Studio dei Tumori
                    ural anticoagulants and selected endothelial markers in    (IRST)”Dino Amadori”, Meldola (FC), Italy; 3Division
                    patients with different severity of covid-19 infection     of Internal Medicine, Hematology Service, Regional
                    during the first wave of the pandemic. Methods: We         Hospital “A. Cardarelli”, Campobasso, Italy
                    retrospectively analyzed antigen levels of d-dimer
                    (DD), protein C (PC), total protein S (PS), total tissue   Background: Several cases of unusual deep vein throm-
                    factor pathway inhibitor (TFPI), antithrombin              botic (DVT) events and thrombocytopenia have been
                    (Diagnostica Stago, France) and of the endothelial         observed after vaccination with Vaxzevria® (ChAdOx-1




         [page 138]                                      [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 139



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    nCov19) from the end of 2020 to early 2021. Vaccine-          Table 1.
                    induced thrombotic thrombocytopenia (VITT) is more
                    common in young females without any relation to hor-
                    monal contraceptives or pre-existing coagulopathy.
                    Symptoms begin from 1 to 2 weeks after the administra-
                    tion of the first dose and are initially mild, simulating a
                    flu-like syndrome, until their worsening leads to hospital-
                    ization. This adverse effect has been shown to occur only
                    after the first dose of Vaxzevria®. Case Report: We here
                    report atypical site DVT associated with severe thrombo-
                    cytopenia after exposure to the first dose of Vaxzevria®.
                    Case 1: In March 2021, a 45-year-old woman, obese,
                    was admitted to the emergency area ten days after the
                    first dose of Vaxzevria® for abdominal pain unrespon-
                    sive to analgesics. Blood tests showed severe thrombocy-
                    topenia and increased D-dimer. A CT scan of the
                    abdomen showed extensive thrombosis of the spleno-
                    mesenteric-portal axis. Low molecular weight heparin
                    (LMWH) was initiated. After one week, treatment




                                                                                                           ly
                    switched to high molecular weight heparin (HMWH).
                    PLT transfusions were given to maintain PLT above




                                                                                                   on
                    30x109/L. Severe hematemesis with acute anemia
                    occurred about 36 hours after the beginning of HMWH;          PO089
                    endoscopy detected hemorrhagic foci in the mucous




                                                                                             e
                    membranes of the pharynx without detectable lesions.          HEPARIN-INDUCED THROMBOCYTOPENIA (HIT)
                    The latest blood tests suggested a pattern of disseminated
                    intravascular coagulation (DIC). A new CT scan con-
                    firmed DVT plus abundant abdominal hemorrhages.
                                                                                      us
                                                                                  AFTER SARS COV 2 INFECTION: A CASE REPORT

                                                                                  F. Pelucchini1, C. Pestelli2, A. Bertelli1, A.A. Rogolino1,
                                                                                  al
                    Therefore, therapy switched to Fondaparinux with high-        F. Cesari1, E. Sticchi1, B. Giusti1, F. Pieralli2, A.M. Gori1,
                    dose intravenous immunoglobulin (IVIGs), and off-label        R. Marcucci1
                                                                          ci

                    use of the anti-IL-6 antibody Tocilizumab. Despite treat-
                    ment, the patient died (Table 1). Case 2: Late in March       1Dept of Experimental and Clinical Medicine, University
                                                                    er


                    2021, a 61-year-old woman was admitted to our center          of Florence, Florence, Italy; 2COVID-19 Intermediate
                    after two weeks after the first dose of Vaxzevria®. The       Care Unit, Careggi University Hospital, Florence, Italy
                                                              m



                    echo-color-doppler showed popliteal and tibial DVT. A
                    CT scan showed left internal jugular vein and bilateral       Introduction: Heparin-induced thrombocytopenia (HIT)
                                                  om




                    pulmonary         embolisms.     Blood      tests    found    is a severe, life-threatening drug reaction associated with
                    moderate/severe thrombocytopenia and increased D-             decrease in platelet count and a high risk of thrombosis
                    dimer. The patient, therefore, started therapy with corti-    caused by platelet-activating antibodies against
                                               -c




                    costeroids, IVIGs, and Fondaparinux. Five days after          PF4/heparin complexes. The atypical clinical and thera-
                    treatment, she experienced a clinical-laboratory improve-     peutic context of the COVID-19 pandemic, with a broad-
                                       on




                    ment. The patient was discharged asymptomatic under           er indication for heparin treatment, could lead to a higher
                    treatment with direct oral anticoagulants (DOACs). In         prevalence of HIT. Here we report a 72 years old woman
                    both cases ADAMTS13 activity, evaluated immediately           operated on for severe aortic valve stenosis, treated with
                                  N




                    after patient admission was normal, thrombophilia             heparin, who, after contracting COVID-19, developed a
                    screening was negative, the anti-PF4-heparin complex          severe HIT. Methods and Results: A 72-year-old
                    immunoassay was negative, autoimmune assay was                woman with a history of hypertension, hiatus hernia,
                    unremarkable (Table 1). Conclusions: According to the         rheumatic polymyalgia and valvulopathy, was admitted
                    available literature on immune response to Adenovirus         for aortic valve replacement surgery. She started heparin
                    and Ad-induced thrombocytopenia, and the correlation          treatment the same day of hospitalization with a platelet
                    between estrogens and thrombotic risk, it can be assumed      count of 202x109/L. After 5 following surgery, platelet
                    that the Adenoviral vector based on Y25 serotype may          count dropped to 83 x109/L. 4T score was 4. Therefore,
                    cause a systemic inflammatory response characterized by       heparin was discontinued and test for anti PF4/heparin
                    a massive platelet activation leading to thrombotic           antibodies was carried out by a chemiluminescent assay
                    thrombocytopenia, especially in women as estrogens            (Acustar Werfen), obtaining a negative result (0.04
                    alone favor the instauration of a prothrombotic state. The    U/mL). As the immunological assay failed to demon-
                    largest studies conducted on VITT demonstrated the            strate the presence of antibodies anti PF4/heparin com-
                    presence of anti-PF4 antibodies in nearly all the patients    plex negative, the patient was again treated with heparin.
                    enrolled. In our 2-cases-experience the assay for the anti-   On the next day, warfarin was added to the therapy and
                    PF4-heparin complex was negative. Early recognition of        the patient tested positive for SARS-COV-2 on nasopha-
                    the symptoms related to disease onset has allowed             ryngeal swab, therefore she was transferred to the
                    prompt and successful management of the disease.              COVID department. The patient was asymptomatic for




                                                               [Hematology Reports 2021; 13:s3]                                                [page 139]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 140



         Poster




                    Covid-19 infection. The day after platelets dropped again      14 patients also had involvement of other districts (3 had
                    to 47 x109/L, so it was decided to stop the heparin treat-     concomitant proximal DVT, 3 had PE, and 10 had CRT,
                    ment and perform again the immunological assay for anti        of which 2 with concomitant PE). Death occurred in 35
                    PF4/heparin antibodies, which resulted positive (>128          (18.9%) patients. There was no relationship between
                    U/mL). Subsequently the HIPA (Heparin Induced                  overall VTE and death (OR 1.2, 95% CI 0.6 - 2.6).
                    Platelet Activation) functional test demonstrated that the     Conclusions: In our cohort we observed a high incidence
                    anti PF4/heparin antibodies detected by the immunolog-         of VTE events. This confirmed the strong relationship
                    ical assay were able to activate platelets. In the following   between COVID-19 and thrombosis, especially in the
                    days after heparin suspension the platelet count returned      critically ill patients. However, we observed no differ-
                    to normal values. One week later, a thrombosis from the        ences in the 30-day mortality rate between VTE and non-
                    basilica vein to the left axillary vein was found, probably    VTE patients. This observation may in part be due to the
                    originating from the peripheral venous catheter.               localization of the index event (mostly distal DVTs or
                    Discussion: We report the case of a 72-year-old woman          CRTs). Although VTE is a common complication of
                    who developed thrombocytopenia after cardiac surgery           COVID-19 infection, its impact on COVID- related mor-
                    with a negative test for PF4/heparin antibodies.               tality is still unclear.
                    Subsequently, concomitant with a positive swab for
                    SARS-COV-2, patient developed anti PF4/heparin anti-
                    bodies, able to functionally activate platelets, thus sug-
                    gesting that SARS-COV-2 infection triggered the forma-




                                                                                                           ly
                    tion of anti PF4/heparin antibodies leading to a clinical




                                                                                                   on
                    presentation of HIT.


                      PO090




                                                                                               e
                    VENOUS THROMBOEMBOLISM-RELATED
                    MORTALITY IN COVID-19 PATIENTS ADMITTED IN
                                                                                       us
                                                                                   Figure 1.
                    INTENSIVE CARE UNIT: A RETROSPECTIVE
                                                                                   al
                    SINGLE-CENTER ANALYSIS
                                                                           ci

                                                                                   PO091
                    G. Turatti, D. Ceccato, F. Boscaro, C. Tirrito,
                                                                      er


                    S. Ragazzo, G. Avruscio                                        ACUTE PORTAL VEIN THROMBOSIS TRIGGERED
                                                                                   BY SARS-COV2 INFECTION
                                                               m



                    U.O.CAngiologia, Azienda Ospedale Università di
                    Padova, Italy                                                  A. Dalla Salda1, A. Soriano2, R. Giacosa2, M. Iotti1,
                                                  om




                                                                                   A. Ghirarduzzi1
                    Background and Aims: The strong association between
                    COVID-19 and venous thromboembolism (VTE) has                  1Cardiovascular Medicine, Department of Internal
                                               -c




                    been widely described, and anticoagulant treatment             Medicine, USL-IRCCS Reggio Emilia; 2Gastroenterology
                    seems to improve overall survival. Otherwise, the effect       and Digestive Endoscopy Unit, USL-IRCCS Reggio
                                       on




                    on severe forms is scarce, and the impact of COVID-            Emilia, Italy
                    related VTE in intensive care unit (ICU)-patients in
                    terms of mortality remains unclear. Our aim was to inves-      Background: Portal vein thrombosis (PVT) is a rare
                                  N




                    tigate the presence and entity of this association in Padua    condition occurring in patients with hepatic cirrhosis,
                    ICU patients. Methods: We retrospectively revised ICU          solid or hematological malignancies or abdominal
                    patients admitted in our university hospital for COVID-        inflammation. Infection by SARS-COV-2 is associated
                    19-related respiratory failure, from January 2020 to April     with respiratory symptoms but also with various throm-
                    2022. All patients underwent a complete Doppler ultra-         botic complications ranging from deep vein thrombosis
                    sound (jugular veins, upper and lower extremities).            /pulmonary embolism to cases of PVT, secondary to a
                    Descriptive data are reported with means and standard          virus induced hypercoagulable state. Notably, patients
                    deviations (SD). Association between VTE and death             affected by hereditary thrombophilia can experience
                    was explored via odds ratio (OR) calculation with 95%          major thromboembolic events during SARS-COV2
                    confidence interval (CI), and a survival analysis was per-     infection, which can require prompt diagnosis and man-
                    formed. Results: We report the exploratory analysis of         agement. Case Report: A 48-year-old male presented at
                    185 patients. Mean age was 65 years and women were             Emergency Department with persistent epigastric pain.
                    slightly underrepresented (65, 35.1%). VTE occurred in         He had recently experienced fever, dry cough and diar-
                    87 (50.3%) patients. Of these, 56 (30.3%) patients had         rhea in relation to a mild symptomatic SARS-COV-2
                    deep vein thrombosis (DVT), respectively 44 distal and         infection and his naso-pharynx SARS-COV-2 swab had
                    15 proximal, 49 (26.5%) patients had catheter-related          resulted negative a week before the onset of abdominal
                    thrombosis (CRT), and 4 (4.6%) patients had pulmonary          symptoms. His medical history was characterized by pre-
                    embolism (PE). Among the 44 patients with distal DVT,          vious splenectomy secondary to an abdominal trauma




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 141



                                                                           XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    and mild gastroesophageal reflux. He had been previous-         Sperimentale e Forense, Università di Pavia, Pavia;
                    ly vaccinated against COVID-19 with one dose of vac-            2Angioematologia, Casa di Cura Lecco “Beato Luigi

                    cine. During Emergency Department visit an urgent               Talamoni”, Lecco, Italy
                    abdomen X-ray resulted normal and the liver ultrasound
                    examination revealed PVT. An urgent abdomen contrast-           Background: The etiology of cerebral venous sinus
                    enhanced computed tomography (CT) confirmed the                 thrombosis (CVST) involves hereditary and acquired
                    PVT showing intra- and extrahepatic portal vein throm-          factors. Acquired factors include brain tumors, brain
                    bosis (24 mm caliber), superior mesenteric vein and             trauma, infections, immune system diseases, pregnancy,
                    splenic vein thrombosis. On laboratory workup, a mild           postpartum complications, trauma, and drugs. Oral con-
                    increase in blood cell count (WBC) and reactive C pro-          traceptives are the most common drugs responsible for
                    tein (RCP) were present, aspartate aminotransferase were        causing CVST. Rapid anticoagulant therapy with low
                    slightly elevated. Medical therapy with low-weight              molecular weight heparin (LMWH) is the preferred
                    molecular heparin and pump protonic inhibitors was              anticoagulant treatment for CVST, and the disease gen-
                    promptly started. During hospital stay, an upper gastroin-      erally has a favourable outcome, with a complete func-
                    testinal endoscopy ruled out portal hypertension-related        tional recovery in about 75% of patients. In some cases,
                    esophageal varices. A thrombophilia workup finally              CVST develops in patients under high dosage progesta-
                    revealed heterozygous prothrombin G20210A mutation.             tive drugs taken for reasons different from contracep-
                    After warfarin bridging, he was discharged home and, a          tion. Recently, new syndrome of vaccine-induced
                    month later, he underwent an abdomen ultrasound exam-           immune thrombotic thrombocytopenia (VITT) has




                                                                                                            ly
                    ination, which confirmed extensive portal vein thrombo-         emerged as a rare side-effect of vaccination against
                    sis with low portal outflow and the presence of collateral      COVID-19. Cerebral venous thrombosis associated




                                                                                                    on
                    circles. He is actually on Warfarin treatment, and he will      with severe hemorrhage is the most common manifes-
                    go on anticoagulant therapy for a minimum of six                tation of this syndrome. The VITT syndrome seemed
                    months. To date, acute thrombotic complications have            more frequent to develop in young woman after the first




                                                                                               e
                    been frequently reported in association with SARS-              dose of vaccines utilizing a non-replicating recombi-
                    COV-2 infection but few data about PVT have been
                    reported in this setting. Nevertheless, the real incidence
                    of this event is perhaps higher than expected, especially
                                                                                        us
                                                                                    nant chimpanzee adenovirus to deliver the genetic
                                                                                    material of the spike protein to the recipient (Vaxzevria,
                                                                                    Oxford–AstraZeneca and Jcovden, Johnson &
                                                                                    al
                    in male patients. Inherited thrombophilia may play a role       Johnson/Janssen). Differently from “classic” CVST,
                    in increasing thrombotic risk in healthy and non-cirrhotic      early reports in which patients were treated with
                                                                            ci

                    patients and a comprehensive thrombophilia screening            heparins described clinical worsening, including death,
                    should be performed. With regard to therapy, the goal of        and early recommendations from health autorities were
                                                                     er


                    treatment is avoiding early and late complications of por-      to avoid heparin because of the resemblance of VITT to
                    tal hypertension, but evidences about the optimal antico-       HIT. Case Reports: We studied the autopsies of two
                                                               m



                    agulant regimen are lacking. Even if limited data are still     young women under estroprogestative treatment cases
                    available in non-cirrhotic patients, either direct oral anti-   of VITT on assignment from the Public Prosecutor
                                                   om




                    coagulants (DOAC) or warfarin can be used.                      Office at the Court. Patient T. F. autopsy diagnosis: tem-
                    Conclusions: Portal vein thrombosis represents a rare           poro-parieto-occipital cerebral hemorrhage following
                    complication of SARS-COV-2 infection. A prompt diag-            thrombosis of the dural venous sinuses (thrombosis of
                                                -c




                    nostic workup is necessary in patients with acute or            the right transverse venous sinus, sigmoid sinus, jugular
                    recent SARS-COV-2 infection complaining acute                   gulf and upper sagittal sinus), occurring 10 days after
                                        on




                    abdominal pain or abdominal symptoms in order to                administration of the first dose of Vaxzevria vaccine in
                    exclude/confirm splanchnic thrombosis. Anticoagulant            subject being treated with estroprogestins. Laboratory
                    treatment is the corner stone of treatment, but the dura-       tests: D-dimer 120,800 μg/L, platelets 35x109/L, pres-
                                   N




                    tion of therapy must be individualized on the clot resolu-      ence of anti-PF4 antibodies. These data allow to classi-
                    tion and recurrent thrombotic risk.                             fy the patient as belonging to the category of certain
                                                                                    VITT syndrome. Patient C.C. autopsy diagnosis: cere-
                                                                                    bral death following massive thrombosis of the venous
                    PO092                                                           sinuses and cortical veins, complicated by widespread
                                                                                    subarachnoid hemorrhage and extensive cerebral intra-
                                                                                    parenchymal infarction occurring a week after adminis-
                    VACCINE-ASSOCIATED CEREBRAL VENOUS                              tration of the first dose of Vaxzevria vaccine. The
                    SINUS THROMBOSIS (CVST) AS A                                    patient started combined estroprogestin with high doses
                    MANIFESTATION OF VACCINE-INDUCED IMMUNE                         didrogesterone and estradiol due to ovarian polycysto-
                    THROMBOTIC THROMBOCYTOPENIA (VITT) IN                           sis and menstrual irregularities. Laboratory tests: D-
                    TWO YOUNG WOMEN. ROLE OF ORAL                                   dimer 34,575 μg/L; platelets 22x109/L, presence of
                    CONTRACEPTIVES                                                  antibodies against PF4. These data allow to classify the
                                                                                    patient also as belonging to the category of VITT syn-
                    C. Franzetti1, L. Tajana1, F. Piovella2                         drome certain. In conclusion, the cases of thrombosis
                                                                                    observed to date during the world vaccination campaign
                    1Unità di Medicina Legale e Scienze Forensi “A.                 are very rare. It is possible to hypothesize that an
                    Fornari”, Dipartimento di Sanità Pubblica, Medicina             inflammatory reaction triggered by the viral vector vac-




                                                                [Hematology Reports 2021; 13:s3]                                             [page 141]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 142



         Poster




                    cine at the level of the endothelium, which seems to be      PO094
                    linked to the generation of antibodies against platelet
                    factor 4 (PF4) in response to the vaccine itself, overlaps
                    with an activation of coagulation factors with conse-        YOU HAVE TO THINK ABOUT IT:
                    quent local thrombosis determined by the presence of         HEMOPHAGOCYTIC LYMPHOHISTIOCYTOSIS
                    estroprogestin drugs.
                                                                                 N. Costantini

                      PO093
                                                                                 Ospedale Giovanni Paolo II, Olbia, Italy

                                                                                 Background: Fever of unknown origin is a challenge
                    ROLE OF THE PROLINE-RICH TYROSINE KINASE                     for internist, because it is particularly confounding and
                    PYK2 IN PLATELET ACTIVATION IN A MURINE                      the underlying disease of the patient not always can
                    MODEL OF ENDOTOXEMIA                                         explain it. There is a strong link between coagulation
                                                                                 and immune system. Case Report: A 78-year-old
                    L. Galgano1,2, M. Vismara2, S.M.G. Trivigno1,2,              woman with rheumatoid arthritis was hospitalised to the
                    S. Rustichelli2, G.F. Guidetti2, M. Torti2, I. Canobbio2     Department of Internal Medicine, in Olbia, in October
                                                                                 2021 for the evaluation of fever. It began one months
                    1Istituto Universitario di Studi Superiori IUSS, Pavia,
                                                                                 earlier, of 39-40°C, which occurred almost daily: she




                                                                                                        ly
                    Italy; 2Dipartimento di Biologia e Biotecnologie             had not joints manifestations and no other symptoms.
                    “Lazzaro Spallanzani”, Università di Pavia, Pavia, Italy     Abatacept, her agent for rheumatoid arthritis, was per-




                                                                                                 on
                                                                                 manently discontinued at admission to the hospital. The
                    Background and Aims: Endotoxemia induced by                  initial laboratory findings revealed: blood cultures were
                    lipopolysaccharide (LPS) in mice leads to thromboin-         always negative; the levels of C-reactive protein and the




                                                                                           e
                    flammation. It is accompanied by hypercoagulability,         erythrocyte sedimentation rate, serologic and immuno-
                    increased platelet and leukocyte activation, and dissem-
                    inated intravascular coagulation [1]. Pyk2 is a non-
                    receptor tyrosine kinase highly expressed in platelets
                                                                                    us
                                                                                 logical analyses were within reference values; elevation
                                                                                 of tumour markers (CA125 150 U/ml; CA15.3 49,1
                                                                                 U/ml), a progressive anemia (hemoglobin 8,3 g/dL),
                    where it controls adhesion, activation, aggregation,         reduction in platelet level up to 101×10³/microL,
                                                                                 al
                    thrombus formation, and platelet-leukocyte/neutrophil        leukopenia (2.9×10³/microl), hypertriglyceridemia
                                                                         ci

                    aggregate (PLA/PNA) formation [2,3]. In this work, the       (triglycerides level up to 563 mg/dL) and marked eleva-
                    role of Pyk2 in platelet and neutrophil activation was       tion in ferritin (>2000 ng/mL ), d-dimer (17129 ng/ml)
                                                                   er


                    investigated in a murine model of endotoxemia.               and lactate dehydrogenase (up to 1357 U/L) without
                    Materials and Methods: LPS (Escherichia coli,                evidence of thrombosis. There was an hypogamma-
                                                             m



                    O111:B4) was injected to induce endotoxemia in WT            globulinemia (IgG 442 mg/dl, IgM 25 mg/dl), hypofib-
                    and Pyk2-KO mice. After four hours, vital parameters         rinogenemia (up to 89 mg/dl), slightly elevated activity
                                                 om




                    were checked, and blood was collected. PLA and PNA           of aminotransferases, increased total bilirubin level and
                    formation, platelet integrin αIIbβ3 activation, P-selectin   beta 2 microglobulin. Echocardiography, gastroscopi
                    expression and neutrophil αMβ2 activation were analysed      and colonoscopy were all unremarkable. By using com-
                    by flow cytometry. Phosphorylation of signalling pro-
                                              -c




                                                                                 puted tomography of the chest and abdomen no lym-
                    teins was analysed ex vivo on isolated platelets in          phadenopathy was recorded, the size of spleen was nor-
                                       on




                    immunoblotting and aggregation was monitored in a            mal but there was a mural thickness in the sigmoid
                    Born lumiaggregometer. Results: LPS-injected Pyk2-           colon with increased density in adjacent adipose tissue:
                    KO mice show a reduced disease score, a more severe          however the intraoperative aspect was normal. Two
                                  N




                    thrombocytopenia, and a lower number of circulating          lines of empirical antibiotic terapie were used, but they
                    PLA/PNA because of a reduced P-selectin exposure on          were then discontinued as infectious workup was
                    platelets compared to WT mice. LPS injection increases       always negative. Bone marrow biopsy revealed also
                    phosphorylation of MAPK and Akt induced by                   reactive alterations. Patient was treated with oral
                    Thrombin Receptor Activating Peptide 4 (TRAP4) in            methylprednisolone, 40 mg per day, with rapid resolu-
                    WT platelets, which is abolished in Pyk2-KO platelets.       tion of fever; it came back on reducing the dose of the
                    No differences in term of aggregation, platelet αIIbβ3 and   steroid. On December the patient was transferred to
                    neutrophil αMβ2 activation were observed in the absence      another department of Internal Medicine, in Cagliari,
                    of Pyk2. Conclusions: Our results demonstrate that           where a new bone marrow biopsy was done, almost two
                    Pyk2 regulates MAPK and Akt phosphorylation, platelet        months after the first: it exhibits typical Chronic lym-
                    P-selectin expression and PLA/PNA formation in a             phocytic leukemia phenotype.The patient met 5 of 8
                    mouse model of endotoxemia, thus modulating the first        HLH-2004 criteria for diagnosis of acquired hemo-
                    trigger of thromboinflammation.                              phagocytic syndrome, including 1 clinical (fever) and 4
                                                                                 laboratory (cytopenia, hyperferritinemia, hypertriglyc-
                    References:                                                  eridemia and hypofibrinogenemia,high levels of soluble
                     1. Shannon, Platelets. 2015;26:302-8.                       IL-2 receptor) criteria. HLH patient had a clinical
                     2. Canobbio et al., Blood. 2013;121:648-57.                 response to systemic dexamethasone and she was
                     3. Momi et al., Haematologica. 2022; in press.              admitted to hematology. Conclusions: HLH is likely




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 143



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    under-recognized and may not be visible until late in        adhesion of monocytes to endothelial cells. Gene
                    disease progression; bone marrow biopsies performed          expression of Vascular and Intercellular adhesion mole-
                    early in the course of secondary disease may be normal,      cules 1 (VCAM-1, ICAM-1), E-selectin (SELE) and
                    so we have to think about it. We must not be satisfied       chemokine CC ligand-2 (CCL2) were evaluated by real
                    with the initial diagnosis. Secondary hemophagocytic         time PCR. VCAM-1 and ICAM-1 membrane exposure
                    lymphohistiocytosis is a potentially life-threatening        was analysed by flow cytometry. Nuclear factor kappa-
                    complication of rheumatic disease, therefore in our          light-chain-enhancer of activated B cells (NF-kB) phos-
                    patient we should not have looked for alternative diag-      phorylation and nuclear translocation were evaluated by
                    noses: instead HLH etiology was ascribed to haemato-         western blot and immunofluorescence staining, respec-
                    logical malignancy. D-dimers levels were highly incre-       tively. Nitric oxide bioavailability through cyclic
                    ate probably for the demonstrated fibrinolytic activity, a   guanosine monophosphate (NO/cGMP) levels was
                    coagulation disorders described in HLH.                      measured by ELISA. Cell migration was performed by
                                                                                 scratch assay and by measuring proteins involved in the
                                                                                 focal adhesion turn-over such as Paxillin and Vinculin.
                    PO095                                                        Vessel formation was evaluated by Matrigel tube for-
                                                                                 mation assay. Results: AM significantly reduced TNF-
                    ENDOTHELIAL CELLS FROM UMBILICAL CORD OF                     α stimulated monocyte-endothelium interaction and the
                                                                                 expression of VCAM-1, ICAM-1, SELE and CCL2 in
                    WOMEN AFFECTED BY GESTATIONAL DIABETES:
                                                                                 C- and GD-HUVECs. That was also reflected on a




                                                                                                        ly
                    AN IN VITRO VASCULAR MODEL TO STUDY THE
                                                                                 reduction on membrane exposure of VCAM-1 and
                    ANTI-INFLAMMATORY AND PRO-ANGIOGENIC




                                                                                                 on
                                                                                 ICAM-1 in both cell types upon AM stimulation. This
                    ROLE OF AMNIOTIC MEMBRANE IN DIABETIC                        was associated with a signiﬁcant decrease of phospho-
                    FOOT ULCER                                                   rylation of NF-kB and its cytoplasm-nucleus transloca-
                                                                                 tion. Finally, AM-increased NO bioavailability was




                                                                                             e
                    C. Pipino1, I. Cappellacci1, P. Di Tomo1,                    affected by eNOS inhibitor L-NAME, suggesting that
                    Á. Bernabé-García2, J. Stelling-Férez2,3,
                    N. Di Pietrantonio1, D. Mandatori1, M.P.A. Baldassarre4,
                    F.J. Nicolás2, A. Pandolfi1
                                                                                    us
                                                                                 AM activation of eNOS could be a possible way of reg-
                                                                                 ulating this phenomenon. Of note, C- and GD-HUVECs
                                                                                 migration capability was ameliorated after incubation
                                                                                 al
                    1Center
                                                                                 with AM in wound healing scratch assay. Moreover, the
                            for Advanced Studies and Technology-CAST (ex         increased number of focal adhesions involving both
                                                                         ci

                    CeSI-MeT), Department of Medical, Oral and                   Paxillin and Vinculin after AM treatment suggests a
                    Biotechnological Sciences, University G. d’Annunzio          rapid turnover of these structures supporting cell migra-
                                                                   er


                    Chieti-Pescara, StemTeCh group, Chieti, Italy;               tion improvement. Furthermore, several angiogenic
                    2Regeneration, Molecular Oncology and TGFß, IMIB-
                                                                                 parameters, analyzed following Matrigel tube forma-
                                                             m



                    Arrixaca, Murcia, Spain; 3Department of Nutrition and        tion assay, were improved in AM stimulated GD-
                    Food Technology, UCAM, Murcia, Spain; 4Center for            HUVECs, thus promoting better interconnected net-
                                                 om




                    Advanced Studies and Technology-CAST (ex CeSI-MeT),          work areas. Conclusions: Overall, these results indicate
                    Department of Medicine and Aging Sciences, University        that in our cell model AM is effective in reducing
                    Gd’Annunzio Chieti-Pescara, Chieti, Italy                    hyperglycemia and improving vasculogenesis, possibly
                                              -c




                                                                                 through the modulation of NO bioavailability, which
                    Background and Aims: Diabetic foot ulcer (DFU) is a          plays a key role in the vascular homeostasis balance.
                                       on




                    severe complication of diabetes which affects 15% of         This study suggests that AM improves chronic wound
                    patients with advanced stages of the disease. Despite        healing by both enhancing angiogenesis and decreasing
                    improvements in the treatment, DFU remains the main          inflammation, thus supporting its clinical application
                                  N




                    cause of lower limb amputation. One of the most rele-        for diabetic foot ulcers.
                    vant diabetic wounds healing issues is the reduced
                    peripheral blood flow, the diminished neovasculariza-
                    tion and chronic inflammation. Amniotic membrane             PO096
                    (AM) has shown promising results in the treatment of
                    DFU. Here, the potential role of AM on endothelial
                    cells isolated from umbilical cords of gestational dia-      INTRACARDIAC THROMBOSIS IN A PATIENT
                    betes affected women (GD-HUVECs) has been investi-           WITH PRIMARY ANTIPHOSPHOLIPID
                    gated. Indeed, GD-HUVECs, considering their epige-           SYNDROME[/B]
                    netic memory and pro-inflammatory phenotype
                    induced by in vivo hyperglycemia during pregnancy,           E. Marrone1, A.M. Cerbone2, D. D’Auria1, F. Gallucci1,
                    could be a proper model to investigate the AM promot-        C. Romano1, A. Tufano2, P. Morella1, G. Di Minno2
                    ing mechanisms in repairing diabetic chronic non-heal-
                                                                                 1InternalMedicine Unit 3, Cardarelli Hospital, Naples,
                    ing wounds. Methods: For Control (C)- and GD-
                    HUVECs, stimulated with low doses of the pro-inflam-         Italy; 2Department of Clinical Medicine and Surgery,
                    matory Tumour Necrosis Factor-α (TNF-α, 1ng/ml), in          Federico II University Hospital, Naples, Italy
                    basal condition and after 24h incubation with AM,
                    monocytes adhesion assay was performed to assess the         Background: Antiphospholipid syndrome (APS) is a




                                                              [Hematology Reports 2021; 13:s3]                                           [page 143]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 144



         Poster




                    systemic autoimmune disorder characterized by venous          Radiologiche ed Ematologiche, Università Cattolica,
                    and arterial thrombosis, recurrent abortion, and throm-       Fondazione Policlinico Universitario AGemelli IRCCS,
                    bocytopenia. Cardiac involvement in APS may be pre-           Roma; 2Servizio Malattie Emorragiche e Trombotiche,
                    sented as right or left ventricular dysfunction, pul-         Fondazione Policlinico Universitario AGemelli IRCCS,
                    monary hypertension, accelerated atherosclerosis,             Roma, Italy
                    occlusive arterial disease (angina, myocardial infarc-
                    tion), valve abnormalities or less frequently as intracar-    Background and Aims: Measurement of D-dimer (D-
                    dial thrombosis and myxoma. Few cases of intracardiac         D) can vary according to different instruments and
                    thrombosis associated with primary APS are described          testing methods; moreover, results can be falsely
                    and the management of these patients is challenging. We       increased by heterophilic antibodies, which exhibit
                    describe a case of intracardiac thrombosis in a young         multispecific reactivity. D-D is persistently increased
                    patient with primary APS. Case Report: A 28-year-old          in 15-20% of pts after COVID-19; conversely, D-D
                    female patient was admitted to our hospital because of        after SARS-CoV-2 vaccination was reported normal.
                    swelling and pain of the right leg. Venous Doppler ultra-     We report a series of pts with D-D increased after
                    sound showed femoral, popliteal and great saphenous           SARS-CoV-2 vaccination. Methods: We investigated
                    vein thrombosis. CT scan showed pulmonary embolism            16 pts (M/F 7/9, median age 64, range 32-76) referred
                    (right pulmonary artery). It was negative for active          for D-D >500 ng/ml (median 1502, range 550-5802)
                    malignancy. Anticoagulant treatment with Fondaparinux         after SARS-CoV-2 vaccination and/or COVID-19
                    (7.5 mg daily) was started. She had never smoked, did         infection; in 11 D-D was >1000 ng/ml. Vaccination




                                                                                                        ly
                    not take any medication, in particular oral contracep-        was adenoviral (Ad) vector-based in 4 pts, mRNA-
                                                                                  based in 10, and heterologous Ad vector-/mRNA-




                                                                                                 on
                    tives. She was not pregnant. Anticoagulant protein S,
                    protein C, and Antithrombin were normal. Factor V             based in 1. Three pts had COVID-19 before vaccina-
                    Leiden and prothrombin G20210A mutation were nega-            tion; one was pregnant and had COVID-19 without
                    tive. A prolonged activated partial thromboplastin time       being vaccinated. Eleven pts were asymptomatic. Five




                                                                                            e
                    (aPTT) (Ratio 4.11, with normal values 0.7-1.3), along        have had vascular complications after vaccination; in
                    with a positive lupus anticoagulant and high titer of anti-
                    cardiolipin (aCL) IgG antibodies at 44 GPL-U/mL (nor-
                    mal range <10 GPL-U/mL) and aβ2 glycoprotein I
                                                                                     us
                                                                                  3 the association with the Pfizer vaccine was doubtful
                                                                                  (deep vein thrombosis=2, ischemic stroke=1), whereas
                                                                                  in 2 the association with the AstraZeneca vaccine was
                                                                                  al
                    (aβ2GPI) IgG antibodies at 2038 U/mL (normal range            likely (hemorrhagic leukoencephalitis=1, vaccine-
                    ≤20 U/mL) suggested for APS. During hospitalization,          induced thrombotic thrombocytopenia with cerebral
                                                                          ci

                    the patient underwent a transthoracic echocardiogram          vein thrombosis=1) (Table 1). Three different assays
                    that revealed the presence of a cardiac mass in the right     measured D-D levels: Innovance (Siemens
                                                                    er


                    atrium, mobile and attached to the interatrial septum.        Healthineers, Erlangen, Germany), Liatest D-Di Plus
                    Differential diagnosis was between thrombi and myxo-          (Stago, Asnières, France), and AcuStar D-D (Werfen,
                                                              m



                    ma. A comprehensive transesophageal echocardiograph-          Milan, Italy). Fibrin and fibrinogen degradation prod-
                    ic examination and cardiac magnetic resonance imaging         ucts (FDPs) were measured by an immunoturbidimet-
                                                  om




                    confirmed a thrombosis in right atrium. Consultation          ric assay (Stago). All tests were performed in plasma
                    with cardiac surgeons ruled out surgical treatment. She       samples not treated and treated with a heterophilic
                    was discharged with oral anticoagulant therapy with           blocking tube reagent (HBT, Scantibodies Laboratory
                                               -c




                    warfarin maintaining an international normalized ratio        Inc, USA); moreover, the Liatest D-Di Plus declares a
                    (INR) between 3 and 3,5. Repeated blood tests after 12        blocking agent in the buffer. Results: D-D using the
                                       on




                    weeks confirmed the diagnosis of APS. At 3-6-12               Siemens assay with HBT diminished from a median of
                    months of follow-up echocardiograms showed a pro-             1467 ng/mL (range 550-5802) to 1043 ng/mL(range
                    gressive reduction of thrombosis, without cardiac com-        190-6098) (p=0.002). Only 2 pts had D-D <500 ng/mL
                                  N




                    plication. Long-term treatment with warfarin and acetil-      after HBT (No. 1 and 5). The Stago assay induced a
                    salicilic acid was recommended. Conclusions: Cardiac          decrease of D-D in comparison with Siemens either
                    manifestations of APS may be associated with increased        without HBT (median 602 ng/mL, range 165-3880,
                    cardiovascular mortality. A prompt and correct identifi-      p<0.0001 vs. Siemens HBT-) and with HBT (median
                    cation of APS is needed.                                      484 ng/mL, range 33-4219, p<0.0001 vs. Siemens
                                                                                  HBT+). The Werfen assay showed in 13 pts a median
                                                                                  D-D of 481 ng/mL (range 140-4679) without HBT
                      PO097                                                       (p=0.0009 vs. Siemens HBT-) and 454 ng/mL (range
                                                                                  154-4515) with HBT (p=0.01 vs. Siemens HBT+). The
                                                                                  Stago and Werfen assays with no HBT showed D-D
                    D-DIMER LEVELS AFTER SARS-COV-2                               <500 ng/mL in 7/16 and 7/13 pts, respectively. HBT
                    VACCINATION CAN BE FALSELY ELEVATED                           induced no difference between the values obtained by
                    WITHOUT MIRRORING TRUE                                        Stago (p=0.66) or Werfen (p=0.19). FDPs were normal
                    HYPERCOAGULABILITY                                            (<5 µg/mL) in 14/16 pts; 9 had D-D <500 ng/mL using
                                                                                  an HBT or a comparative system, and 5 had elevated
                    V. De Stefano1, S. Lancellotti2                               D-D by all systems but without increased FDP. The
                                                                                  remaining 2 pts had elevated D-D accompanied by
                    1Sezione   di Ematologia, Dipartimento di Scienze             FDPs double the normal (No. 3 and 16): both cases




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 145



                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    had a clinical condition possibly associated with true     Background and Aims: Acute immune-mediated
                    hypercoagulability (aortic prosthesis in pt. No. 3 and     thrombotic thrombocytopenic purpura (iTTP) is a rare
                    pregnancy at term in pt. No. 16). Notably, the patient     but life-threatening thrombotic microangiopathy. In
                    with VITT after adjustment for age had normal D-D          2018 caplacizumab, an anti-von Willebrand factor
                    and FDPs. Conclusions: D-D measurement can be not          nanobody, was approved by the European Medicines
                    reliable after SARS-CoV-2 vaccination, and falsely         Agency for the treatment of acute iTTP. To date, data
                    elevated levels can be demonstrated in half of pts by      stemming from real-world experience are limited. This
                    blocking heterophilic antibodies. However, in all pts      study aims to describe the efficacy and safety endpoints
                    true hypercoagulability is not demonstrable. Such          of caplacizumab treatment in Italian iTTP patients from
                    results should prompt to use of confirmatory laborato-     a real-world clinical setting. Methods: In this observa-
                    ry systems to avoid undue anxiety and unuseful and         tional retrospective study, 26 patients from the Milan
                    costly examinations in asymptomatic pts with falsely       TTP Registry receiving caplacizumab treatment for an
                    elevated D-D after SARS-CoV-2 vaccination.                 acute iTTP episode between 2018 and August 2021
                                                                               were enrolled. Clinical signs/symptoms at the time of
                    Table 1.                                                   onset were categorized as follows: systemic (fatigue,
                                                                               fever, abdominal pain, headache, vomiting, jaundice);
                                                                               neurological (ischemic stroke, transient ischemic
                                                                               attack, epileptic seizures, coma, personality disorders,
                                                                               focal neurological signs); hemorrhagic (skin bleeds,




                                                                                                      ly
                                                                               mucosal bleeds, hematuria, meno-metrorrhagia, gas-
                                                                               trointestinal hemorrhages); cardiovascular (acute coro-




                                                                                               on
                                                                               nary syndrome, myocardial infarction, electrocardio-
                                                                               graphic ischemic changes); renal (acute renal failure,
                                                                               anuria, need for dialysis, emoglobinuria, proteinuria,




                                                                                         e
                                                                               oliguria). Clinical response was defined as the sus-
                    PO098                                                         us
                                                                               tained normalization of platelet count (≥150,000/µL)
                                                                               for at least two consecutive days. Exacerbation was
                                                                               defined as a platelet count decrease (to less than
                    CAPLACIZUMAB FOR THE MANAGEMENT OF
                                                                               al
                                                                               150,000/µL) occurring after a clinical response and
                    ACUTE IMMUNE-MEDIATED THROMBOTIC                           before a clinical remission within 30 days from stop-
                                                                        ci

                    THROMBOCYTOPENIC PURPURA: REAL WORLD                       ping PEX or caplacizumab. Partial ADAMTS13 remis-
                    DATA FROM THE MILAN TTP REGISTRY                           sion was defined when ADAMTS13 activity was
                                                                  er


                                                                               ≥20%. Total ADAMTS13 remission was defined as
                    P. Agosti1, P. De Leo2, M. Capecchi2,3, A. Artoni2,        ADAMTS13 activity ≥45%. Results: The main patient
                                                             m



                    I. Mancini1, M. Biganzoli2, B. Ferrari2, S. Gattillo4,     characteristics and acute event features are shown in
                    S.M. Trisolini5, E. Rinaldi6, G. Podda7, L. Prezioso8,     Table 1. All patients were treated with plasma-
                                                 om




                    S. Prassede9, L. Facchini10, D. Caramazza11,               exchange (PEX), corticosteroids and caplacizumab,
                    G. Tolomelli12, F. Peyvandi1,2                             except for one patient with mild thrombocytopenia
                                                                               (113000/µL) treated with steroids and caplacizumab.
                                              -c




                    1Università   degli Studi di Milano, Department of         Rituximab was used in 16/24 patients (66.7%).
                    Pathophysiology and Transplantation, and Fondazione        Caplacizumab was started a median of 1 day after PEX
                                      on




                    Luigi Villa, Milan, Italy; 2Fondazione IRCCS Ca’           initiation (Table 1). Thirty percent of patients started
                    Granda Ospedale Maggiore Policlinico, Angelo               caplacizumab 7 days or more after PEX start to treat
                    Bianchi Bonomi Hemophilia and Thrombosis Center,           refractory TTP or exacerbation. All patients achieved
                                  N




                    Milan, Italy; 3Università degli Studi di Milano,           clinical remission. In patients treated with caplacizum-
                    Department of Biomedical Sciences for Health, Milan,       ab within 24 hours after PEX start, the median time to
                    Italy; 4Immuno-Hematology and Transfusion Medicine         platelet count normalization was 3.5 days (3.0-4.3),
                    Unit, San Raffaele Hospital, Milan, Italy; 5Hematology,    with a median of 6 days of PEX (4.3-7.8) to achieve
                    Department of Translational and Precision Medicine,        clinical response. One patient experienced acute TTP
                    “Sapienza” University of Rome, Rome, Italy;                exacerbation 11 days after stopping caplacizumab. Two
                    6Hematology Unit, APerrino Hospital, Brindisi, Italy;      caplacizumab-related adverse events leading to drug
                    7U.OMedicina III, ASST Santi Paolo e Carlo,                discontinuation were observed, an episode of hemopty-
                    Dipartimento di Scienze della Salute, Università degli     sis treated with a plasma-derived von Willebrand fac-
                    Studi di Milano, Milan, Italy; 8Haematology and BMT        tor/factor VIII-containing concentrate and artery
                    Unit, Azienda Ospedaliero-Universitaria di Parma,          embolization, and a gastrointestinal bleeding requiring
                    Parma, Italy; 9Ospedale Civile “Spirito Santo,”            blood transfusion in a patient with several conditions
                    Pescara, Italy; 10Hematology Unit, Azienda Unità           predisposing to bleeding. Conclusions: With a median
                    Sanitaria Locale-IRCCS di Reggio Emilia, Reggio            of 4 days to achieve platelet count normalization,
                    Emilia, Italy; 11Hematology, University Hospital           caplacizumab proved effective and safe for the treat-
                    “Ospedale di Circolo e Fondazione Macchi” - ASST           ment of acute iTTP in the real-world clinical setting of
                    Sette Laghi, Varese, Italy; 12Unit of Hematology, Rimini   the Milan TTP Registry, consistently with the results of
                    Hospital “Infermi”, Rimini, Italy                          randomized clinical trials.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 146



         Poster




                    Table 1.                                                  Among patients with lymphoma, the most common
                                                                              type was diffuse large B-cell lymphoma (n=191), fol-
                                                                              lowed by mantle cell lymphoma (n=19), peripheral T
                                                                              cell lymphoma (n=10), Burkitt lymphoma (n=4), and
                                                                              primary mediastinal large B-cell lymphoma (n=3).
                                                                              Results: Among 227 patients with diagnosis of
                                                                              aggressive lymphoma, the incidence of VTE was
                                                                              19.8% (n=45) in a median follow up of months 28.87
                                                                              (range, 0.23-177.18). Only about half of patients
                                                                              (55.5%) were treated with thromboprophylaxis. VTE
                                                                              events occurred in the older patients (pV=0.029), with
                                                                              concomitant HIV infection (pV=0.031), and with
                                                                              spleen involvement (pV=0.022). At diagnosis, higher
                                                                              level of LDH (pV=0.042), leukocytosis (pV=0.004)
                                                                              and higher neutrophil-to-lymphocyte ratio (NLR)
                                                                              (pV=0.022) are risk factor of development of VTE.
                                                                              While the VTE did not significantly affect the overall
                                                                              survival, it caused increased morbidity with an inci-
                                                                              dence of recurrent VTE of 9%. During a follow-up




                                                                                                     ly
                                                                              period of 23.09 month (range,1.01-135.95), 46 of 231




                                                                                              on
                                                                              patients (19.9%) with diagnosis of multiple myeloma
                                                                              experienced a VTE. Twenty-three (50%) events were
                                                                              TVP, followed by EP in 28% and TVS in 22% of
                                                                              patients. There was no effect of clinical and laboratory




                                                                                        e
                                                                              features at diagnosis on the occurrence of thrombosis,
                                                                                 us
                                                                              with the exception of older age (pV=0.027), bone dis-
                                                                              ease (pV=0.002), and the presence of central venous
                                                                              catheter (pV<0.001). Interesting, treatment of myelo-
                                                                              al
                                                                              ma patients with Lenalidomide-based therapy,
                                                                              Carfizomib-based therapy and monoclonal antibody-
                                                                      ci

                      PO099                                                   based therapy increase the risk of VTE (HR 2.55
                                                                              (1.31-4.95), pV=0.005; HR 6.1 (2.8-13.6), pV<0.001;
                                                                er


                    THE INCIDENCE AND RISK FACTORS OF                         HR 2.36 (1.07-5.19), pV=0.037; respectively).
                    THROMBOSIS IN AGGRESSIVE LYMPHOMA AND                     Conclusions: VTE is a frequent complication in
                                                           m




                    MYELOMA PATIENTS: A 6-YEAR SINGLE-CENTER                  patients with aggressive lymphoma and multiple
                    EXPERIENCE                                                myeloma. In our cohort of aggressive lymphoma
                                                om




                                                                              patients, the new inflammatory status markers NLR is
                    E. Morsia, E. Torre, F. Marrara, S. Morè, S. Cacciagiù,   an independent risk factor for VTE. However, in
                    V.M. Manieri, G. Guidi, M. Offidani, A. Olivieri,         patients affected by multiple myeloma the therapeutic
                                             -c




                    S. Rupoli                                                 approach seems play a crucial role in development of
                                                                              thrombosis. Emerging clinical and therapeutic risk
                                      on




                    Clinica di Ematologia, Ospedali Riuniti di Ancona,Italy   factors were found, but further studies are needed to
                                                                              develop a predictive model for identifying patients
                                                                              with lymphoma and myeloma who are at high risk for
                                 N




                    Background and Aims: The risk of venous throm-
                    boembolism (VTE) in patients with cancer is higher        developing thrombosis and for planning a tailored pro-
                    compared to healthy people. Recent studies demon-         phylactic strategy.
                    strated a high incidence of VTE in patients with hema-
                    tologic malignancies as well as in patients with solid
                    cancers. Although some, very well validated scores        PO100
                    delineate the risk of VTE by cancer subtype and other
                    risk factors, hematologic malignancies are less repre-    COMPARISON OF STANDARD VS. LOW DOSE
                    sented in these models. Moreover, this setting of         DIRECT ORAL ANTICOAGULANTS FOR THE
                    patients represents a peculiar entity that undergoes      SECONDARY PROPHYLAXIS OF VENOUS
                    therapy that can be thrombogenic. The aim of our          THROMBOEMBOLISM IN PATIENTS AFFECTED BY
                    study was to analyze VTE incidence and factors relat-     HEMATOLOGIC MALIGNANCIES
                    ed to thrombosis risk, in a retrospective cohort of
                    patients with aggressive lymphoma and multiple            F. Malfona, A. Serrao, G.M. Assanto, S. Olivieri,
                    myeloma. Methods: We reviewed the medical records         S. Ligia, C. Santoro, A. Chistolini
                    of 227 patients diagnosed with aggressive lymphoma
                    and 231 patients with multiple myeloma who referred       Hematology, Department of Translational and Precision
                    to our institute from January 2015-December 2020.         Medicine, Sapienza University, Rome, Italy




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                                                                      XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Background: Venous thromboembolism (VTE) is one           PO101
                    serious complication in patients with hematologic
                    malignancies due to the release of procoagulant fac-
                                                                              THE SAFETY AND EFFICACY OF DIRECT ORAL
                    tors by neoplastic cells, central venous catheter and
                    chemotherapy. Considering that hematologic malig-         ANTICOAGULANTS FOR ATRIAL FIBRILLATION IN
                    nancies represent a non-transient VTE risk factor, the    PATIENTS AFFECTED BY HEMATOLOGIC
                    treatment and secondary prophylaxis of VTE are an         MALIGNANCIES: A SINGLE-CENTER EXPERIENCE
                    important step in the therapeutic approach of hemato-
                    logic patients. Although direct oral anticoagulants       F. Malfona, A. Serrao, G.M. Assanto, S. Ligia,
                    (DOACs) demonstrated their efficacy and safety in the     E. Baldacci, C. Santoro, G.O. Chavez, A. Chistolini
                    treatment of acute phase of VTE cancer related, there
                    are few data in the setting of the secondary prophylax-   Hematology, Department of Translational and Precision
                    is. Aims: The aim of our study was to evaluate the        Medicine, Sapienza University, Rome, Italy
                    efficacy and safety of DOACs in the VTE secondary
                    prophylaxis in hematologic patients, comparing full       Background and Aims: Pre-existing atrial fibrillation
                    and low dose of DOACs. Methods: This retrospective        (AF) is not uncommon in cancer patients receiving
                    monocentric study collected data of patients affected     active therapy, and the risk of AF increases during the
                    by hematologic malignancies with VTE (pulmonary           period adjacent to diagnosis and during tratment.
                    embolism and deep vein thrombosis of typical sites)       Managing anticoagulation in onco-hematological




                                                                                                     ly
                    treated with DOACs. We divided the patients in 2          patients with atrial fibrillation is a clinical challenge
                                                                              considering the risk of bleeding and thrombosis with




                                                                                              on
                    groups according to DOACs dose performed after the
                                                                              limited data, mainly concerning low molecular weight
                    first 6 months of VTE treatment: one group continue
                                                                              heparin. The aim of our study was to evaluate the effi-
                    with standard dose and the other group shifted to apix-
                                                                              cacy and safety of direct oral anticoagulants (DOACs)
                    aban 2.5 BID or rivaroxaban 10 QD. Results: We




                                                                                        e
                                                                              in patients affected by hematologic malignancies with
                    studied 55 patients (Table 1). After a median time of 6
                    months from a first VTE, 29 patients continued with
                    DOAC at full dose, 26 patients shifted to DOACs at
                    reduced dose. After a median time of 24 months
                                                                                 us
                                                                              AF. Methods: A single-center retrospective cohort
                                                                              study was performed by reviewing medical records of
                                                                              patients with a confirmed diagnosis of hematologic
                                                                              al
                                                                              malignancy who presented a pre-existing AF or devel-
                    (range 19-32) no thrombotic recurrences were              oped AF during the clinical course and treated with
                    observed in the two groups. We reported 4 minor
                                                                      ci

                                                                              DOACs. Oral anticoagulation was continued until a
                    bleeding events: 3 in the group treated with full dose    platelet count ≥50x109/l. Results: We included 97
                                                                er


                    and 1 in the other group. We did not report any statis-   patients (Table 1). DOACs therapy was performed for
                    tically significant differences in terms of DOACs effi-   a median time of 25 months (range 10-108 months).
                    cacy and safety between the two groups. Conclusions:
                                                           m



                                                                              We did not observe any thrombotic adverse event dur-
                    Our experience supports the use of DOACs also at          ing the follow-up. We observed 14 (14.4%) bleeding
                                                      om




                    reduced dosage in the VTE secondary prophylaxis in        complications: 1 major, 12 clinically relevant non
                    patients affected by hematologic malignancies.            major and 1 minor hemorrhagic event. The major
                                                                              complication was a severe gastrointestinal bleeding in
                                                   -c




                                                                              a patient affected by non-Hodgkin lymphoma during
                    Table 1. Patients’ characteristics.
                                                                              local disease progression: in this case, DOAC was
                                          on




                                                                              definitively discontinued. The CRNM bleedings were
                                                                              5 gastrointestinal haemorrhages and 7 hematomas: in
                                                                              three of the above cases DOAC was temporarily
                                    N




                                                                              replaced with LMWH (median time 8 days) because
                                                                              of thrombocytopenia with a platelet count between
                                                                              100X109/L and 50X109/L; in one case DOAC was
                                                                              definitively discontinued in one patient with a platelet
                                                                              count <20X109/L. The minor event was a mild epis-
                                                                              taxis, for which no DOAC withdrawal was needed. In
                                                                              9 cases DOAC was temporary discontinued for a
                                                                              median time of 21 days (range 5-28): 7 for severe tran-
                                                                              sient thrombocytopenia and 2 for chemotherapy-relat-
                                                                              ed coagulopathy. Four/14 (28.6%) bleeding complica-
                                                                              tions (1 major and 3 CRNM) were reported during the
                                                                              concomitant administration of BTK inhibitor ibruti-
                                                                              nib. No statistically correlation was found between
                                                                              bleeding complications and sex (p=0.129), age
                                                                              (p=0.264), type of DOAC (p=0.527), hematologic dis-
                                                                              ease (p=0.729) stage of the hematologic disease
                                                                              (p=0.405). Conclusions: Our study evaluated the effi-
                                                                              cacy and safety of DOACs in patients with AF and




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 148



         Poster




                    hematologic neoplasms. In this setting, in light of the     1Clinicadi Ematologia, Ospedali Riuniti di Ancona;
                    recognized increase of bleeding risk for concomitant        2ClinicaNeurologica, Ospedali Riuniti di Ancona;
                    thrombocytopenia, coagulopathy and cancer treat-            3Medicina di Laboratorio, Ospedali Riuniti Ancona,

                    ment, clinical surveillance is required even in patients    Italy
                    at high thromboembolic risk. In our study, we confirm
                    the higher incidence of hemorrhagic adverse events          Background and Aims: Cerebral vein thrombosis
                    versus thrombotic complications. At univariate analy-       (CVT), including thrombosis of cerebral veins and major
                    sis, none of the variables evaluated were statistically     dural sinuses, is a rare disorder accounting for <1% of all
                    significant, even considering the different type of anti-   strokes. It is three times more common in younger women
                    cancer therapies: these results could be related to our     as compared to men. CVT can appear with a variety of
                    small sample size. In patients with hematologic malig-      symptoms including neurological deficits and can
                    nancies and AF, specific monitoring modalities should       progress to hemorrhage. Patients affected by myeloprolif-
                    be considered. In addition to careful clinical examina-     erative neoplasms (MPNs) including polycythemia vera,
                    tion for bleeding signs and regular monitoring of liver     essential thrombocythemia, and primary myelofibrosis
                    and renal function, repetitive full blood counts should     are prone to develop thrombosis, especially in unusual
                    be evaluated. Our data support the efficacy of DOACs        sites, either at diagnosis or during follow-up. In these
                    in the treatment of AF in patients with hematologic         patients the prevalence of CVT is 1% and can be the first
                    malignancies but suggest caution in case of thrombo-        clinical manifestation. The aim of this study is to analyze,
                    cytopenia and disease progression.                          in a retrospective cohort of CVT patients, the prevalence,




                                                                                                         ly
                                                                                clinical features and risk factors for thrombosis.
                    Table 1. Patients’ characteristics.                         Additionally, this study shows the clinical outcome of




                                                                                                 on
                                                                                CVT, with a specific focus on CVT in MPN patients.
                                                                                Methods: The patient cohort included 38 patients of our
                                                                                center who experienced CVT during a 20-year period




                                                                                           e
                                                                                (January 2002-December 2021). Diagnosis of CVT was

                                                                                    us
                                                                                established with CT or MRI. MPN are defined according
                                                                                to WHO 2016 criteria. Data regarding demographic, clin-
                                                                                ical and radiological features were retrospectively collect-
                                                                                al
                                                                                ed from medical records. Results: Our cohort includes 38
                                                                                patients with CVT (median age 42 years, range 20-79;
                                                                          ci

                                                                                65% females) in median follow up of 21.35 months
                                                                                (range, 9,13-255,38). Comprehensive thrombophilia test-
                                                                   er


                                                                                ing was performed in most cases (94%), showing pres-
                                                                                ence of prothrombin G20210A (10.5%), factor V Leiden
                                                             m



                                                                                (7.9%), and hyperhomocysteinemia (26.3%) as the three
                                                                                most common abnormalities. Other risk factors observed
                                                      om




                                                                                were oral contraceptive and mastoiditis in 31.6% and
                                                                                15.8% of patients, respectively. Notably, one patient
                                                                                received the ChAdOx1-S vaccine 10 days prior to CVT.
                                                   -c




                                                                                VTE events occurred most frequently in multiple sites,
                                                                                involving transverse, sigmoid and sagittal sinuses in 65%
                                          on




                                                                                of cases. The most common presenting symptom of CVT
                                                                                was headache (81.6%) followed by visual loss (36.8%).
                                                                                Post diagnosis, 34 patients received antithrombotic treat-
                                    N




                                                                                ment with vitamin K antagonists (VKA). Twenty-five
                                                                                patients (66%) achieved CVT recanalization within a
                                                                                median time of 5.6 months from the appearance of the
                                                                                thrombosis and 5 patients (13%) had recurrent venous
                                                                                thrombosis. However, CNS hemorrhage occurred in three
                                                                                patients (7.9%) and menorrhagia in one. In total, 6
                                                                                patients with CVT (median age 34 years, range 21-50)
                                                                                were also diagnosed with MPN JAK2 mutated, which in
                                                                                half cases occurred simultaneously with the thrombosis.
                      PO102
                                                                                All these patients were treated with hydroxyurea plus
                                                                                VKA. Two patients had prior VTE in median 11.5 years
                    THE CLINICAL PATTERNS AND ETIOLOGICAL                       before MPN while in two patients a recurrent event
                    FEATURES OF CEREBRAL VEIN THROMBOSIS IN                     occurred. Except for smoke (pV=0.048) and provoked
                    SINGLE-CENTER STUDY                                         thrombosis due to mastoiditis (pV=0.046), no significant
                                                                                difference was found in term of thrombophilia abnormal-
                    E. Torre1, E. Morsia1, F. Marrara1, A. Fiorentini1,         ities and other risk factors for thrombosis comparing CVT
                    A.R. Scortechini1, M. Silvestrini2, L. Da Lio3,             with and without MPN. Moreover, patients with MPN
                    A. Olivieri1, S. Rupoli1                                    and CVT presented at time of thrombosis with higher




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 149



                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    platelet (pV=0.001) and hematocrit (pV=0.022) levels,          1Medicina  Interna, Ospedale di Borgo San Lorenzo,
                    compared to CVT patients without MPN. Finally, within          Firenze; 2Fondazione Arianna Anticoagulazione,
                    this setting of patients factor V Leiden and prothrombin       Bologna; 3Dipartimento di Medicina Sperimentale e
                    G20210A were absent. Conclusions: Our data suggest             Clinica, Università degli Studi di Firenze; 4Dipartimento
                    the clinical importance of ensuring, using JAK2 mutation       di Medicina di Laboratorio, Centro Emostasi e trombosi,
                    screening, the absence of underlying MPN especially in         ASST Cremona, Cremona; 5UO Laboratorio Analisi
                    young patients with unprovoked CVT (without smoke              Distretto Ovest, ULSS 8Berica, Arzignano (VI); 6UO
                    attitude) and blood tests abnormalities.                       Medicina Interna, Presidio Ospedaliero Francesco
                                                                                   Lastaria di Lucera - ASL Foggia, Lucera (FG);
                                                                                   7Department of Clinical, Internal, Anesthesiological and

                    PO103                                                          Cardiovascular Sciences, Sapienza University of Rome,
                                                                                   Roma; 8SOD Malattie Aterotrombotiche, Azienda
                    THROMBOSIS DURING ANTICOAGULANT                                Ospedaliero Universitaria-Careggi, Firenze, Italy
                    THERAPY. THINK OUTSIDE THE BOX
                                                                                   Background and Aims: Direct oral anticoagulants
                                                                                   (DOACs) have been marketed in 2 doses. When used
                    D. Tonello, L. Filippi, L. Cecchetto, M. Marchetti
                                                                                   for stroke prevention in atrial fibrillation (AF) the
                                                                                   dosage of each drug should be chosen following the
                    UOC Medicina Generale, Ospedale Alto Vicentino,
                                                                                   European Medicines Agency indications. However,




                                                                                                          ly
                    Azienda Ulss 7-Pedemontana (VI), Regione Veneto, Italy
                                                                                   inappropriate dose prescrition have been reported with
                                                                                   a frequency ranging from 10% to 57% of cases. The




                                                                                                   on
                    Male, 76-year-old, in November 2016 develops unpro-
                                                                                   purpose of the study is to evaluate the appropriateness
                    voked common femoral DVT during therapy with apixa-
                    ban for atrial fibrillation. Blood tests, neoplastic markers   of the DOACs dosage and the occurrence of ischemic
                                                                                   and haemorrhagic events and death in relation to dose




                                                                                               e
                    and thrombophilia are negative. CT scans and endoscop-
                    ic tests are negative for occult neoplasms. Subsequent         prescription in AF patients enrolled in the observation-
                    checks show no recanalization of DVT. Edema in the
                    lower limb does not regress. In November 2018, throm-
                    bosis shows expansive growth and internal vascularity.
                                                                                      us
                                                                                   al, prospective START2-Register – Survey on
                                                                                   anTicoagulated pAtients RegisTer – an independent,,
                                                                                   ongoing database aimed at recording the history of
                                                                                   al
                    MR angiography and PET-scan confirm a neoplastic               patients starting anticoagulant treatment whatever the
                    intravascular lesion. The histology concludes for a            indication and the drug. Methods: We investigated the
                                                                           ci

                    venous leiomyosarcoma. Male, 64 years old, in August           clinical characteristics of patients receiving therapy
                    2020, develops unprovoked iliac-femoral DVT. Blood             with full, appropriate and inappropriate dosing, and
                                                                     er


                    tests, neoplastic markers and thrombophilia are negative.      with reduced, appropriate and inappropriate dosing for
                    CT scan excludes occult neoplasms or pulmonary                 all the available drugs. Thrombotic events, major and
                                                               m




                    embolism. Discharged on rivaroxaban therapy. In May            clinically relevant not major bleeding events and death
                    2021 there is no recanalization of DVT, but sharp              occurred in the follow-up were recorded. Results: The
                                                  om




                    increase in the edema of the thigh which poses clinical        population cohort included 2106 patients treated with
                    suspicion of recurrence of thrombosis. Blood tests, neo-       DOACs, 53.5% of them were on full dose. Compared to
                    plastic markers, thrombophilia again negative. New CT          patients on full dose, patients on low dose have a higher
                                               -c




                    scans are unchanged. Endoscopic tests for cancer are           frailty profile, with increased number of associated dis-
                    negative. In June 2021 thrombosis shows expansive              eases and more were frequently treated with associated
                                       on




                    growth and arterial vascularization. MRI and PET scan          antiplatelet drugs. The full dose is appropriate in 98.1%
                    confirm the suspicion of intravascular neoplasia.              of cases while the low dose is appropriate in 63.1% of
                    Histological examination diagnoses a synovial sarcoma.         cases. Inappropriated reduced dose prescrition was
                                  N




                    Conclusions: Intravascular malignant tumors can mimic          found in 19.7% of patients on edoxaban, 20.1% of
                    a DVT. Expansive growth, arterial vascularization and          patients on dabigatran, 39.7% of patients on rivaroxa-
                    resistance to anticoagulant therapy can guide us in the        ban, and 63.1% of patients on apixaban. Instead, the
                    diagnostic suspicion.                                          appropriateness of full dose prescription was similar
                                                                                   between the different drugs. In the follow-up there is no
                                                                                   statistically significant difference between thrombotic
                    PO104
                                                                                   and haemorrhagic events in patients treated with appro-
                                                                                   prited or not approprited full and low dose. Instead,
                                                                                   patients treated with inappropriate dosage showed high-
                    ORAL ANTICOAGULANT THERAPY IN ATRIAL                           er mortality rate [Relative Risk 1.6 (1.1-2.3), p=0.01].
                    FIBRILLATION. ANALYSIS OF THE DOSING                           Conclusions: In clinical practice, the use of off-label
                    APPROPRIATENESS OF DIRECT ORAL                                 dosing DOACs is quite common, especially for low
                    ANTICOAGULANTS FROM THE START2 REGISTRY                        doses. We found that AF patients treated with off-label
                                                                                   low doses of DOACs have an increase in mortality rate.
                    L. Ciucciarelli1, E. Antonucci2, F. Crudele3,                  Instead, on the contrary of what expected, the wide use
                    M. Berteotti3, S. Testa4, A. Toma5, L. Masciocco6,             of low dose DOACs is not associated with an increase
                    D. Pastori7, P. Pignatelli7, G. Palareti2, R. Marcucci3,       in thrombotic events, nor in a reduction of bleeding
                    D. Poli8                                                       events.




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 150



         Poster




                                                                              1Azienda      Ospedaliero-Universitaria      Policlinico
                      PO105
                                                                              Umberto 1, Roma; 2Department of General Surgery,
                                                                              Surgical Specialties and Organ Transplantation “Paride
                    LOW DOSE RIVAROXABAN IN PATIENTS WITH                     Stefanini,” Sapienza University, Rome; 3Department of
                    CARDIOVASCULAR DISEASE: A SYSTEMATIC                      Clinical Internal, Anesthesiological and Cardiovascular
                    REVIEW AND META-ANALYSIS OF RANDOMIZED                    Sciences, Sapienza University, Rome; 4Department of
                    CONTROLLED TRIALS                                         Clinical, Internal, Anesthesiological and Cardiovascular
                                                                              Sciences, Sapienza University, Rome, ITALY
                    T. Bucci1, F. Del Sole2, G. Galardo1, F.G. Biccirè1,
                    A. Farcomeni3, P. Pignatelli2, D. Pastori2                Background: Left ventricular thrombosis (LVT) is a
                    1Department
                                                                              life-threatening condition responsible for a high risk of
                                   of General Surgery, Surgical Specialties   stroke and systemic embolism. The aim of our study is
                    and Organ Transplantation “Paride Stefanini,”             to evaluate the effectiveness and safety of vitamin k
                    Sapienza University of Rome, Rome, Italy; 2Department     antagonists (VKAs) in patients with LVT. Methods: All
                    of Clinical, Internal Medicine, Anaesthesiological and    consecutive patients were retrospectively included from
                    Cardiovascular Sciences, Sapienza University of Rome,     January 2013 through January 2021 if they received
                    Rome, Italy; 3Department of Economics and Finance,        VKAs for at least three months. Exclusion criteria com-
                    University of Rome “Tor Vergata”, Rome, Italy             prehended different treatment regimens and unavail-
                                                                              ability of relevant data. Primary outcomes included on-




                                                                                                     ly
                    Background: Low-dose rivaroxaban (LDR) has been           treatment thrombus resolution and major bleedings
                    recently approved as secondary prevention strategy in     while secondary outcomes on-treatment acute ischemic




                                                                                              on
                    patients with coronary artery disease (CAD) and           stroke, acute peripheral embolism, acute myocardial
                    peripheral artery disease (PAD). Pooled data on the       infarction, and clinically relevant non-major bleedings.
                    risk-benefit profile of using LDR in these patients are   Follow-up time was calculated from LVT diagnosis up




                                                                                         e
                    not available. Methods: We performed a systematic         to outcomes development or 12-months follow-up,
                    review and meta-analysis of randomized controlled tri-
                    als (RCT) including CAD or PAD patients treated with
                    LDR. Studies were divided into three groups:1)LDR
                                                                                 us
                                                                              whichever came first. The frequency of thrombus reso-
                                                                              lution was expressed as cumulative incidence with 95%
                                                                              confidence intervals (CIs) while the frequency of other
                    2,5 mg bid+aspirin (ASA) or P2Y12 inhibitor vs ASA
                                                                              al
                                                                              outcomes was descriptively reported. Results: A total
                    and/or P2Y12 inhibitor,2)LDR 2,5 mg bid+ASA vs            of 21 patients were included in the analysis. Baseline
                    ASA,3)LDR 5 mg bid+ASA vs ASA. Cardiovascular
                                                                           ci

                                                                              patients’ characteristics were reported in Table 1.
                    events (CVEs) and major bleeding were the endpoints.      Median age was 73 year and 90.5% of patients were
                                                                  er


                    Number needed to treat (NNT) and number needed to         male. Risk factor for LVT was acute myocardial infarc-
                    harm (NNH) were also calculated. Results: We includ-      tion in 95.2% and primitive dilated myocardiopathy in
                    ed a total of 59,745 patients. Overall, 5,060 CVEs were
                                                             m



                                                                              4.8% of patients. All patients received warfarin with a
                    registered. 7 RCT on LDR were identified. LDR 2,5 mg      median time in therapeutic range (TTR) of 59% and
                    bid was associated with a reduction of CVEs
                                                 om




                                                                              52.4% of patients received concomitant dual
                    (HR:0.84,95%CI:0.77-0.92) with a higher major bleed-      antiplatelet therapy. Cumulative incidences of complete
                    ing risk (HR:1.27,95%CI:1.09-1.47). When we               thrombus resolution at 3-months and 12-months were
                    analysed only the RCT with LDR + ASA, the LDR 2,5
                                              -c




                                                                              42.9% (95% CI, 21.3% to 62.9%) and 57.1% (95% CI,
                    mg twice was associated with a greater reduction of
                                                                              32.8% to 75.5%), respectively (Figure 1). Between
                    CVEs (HR:0.79,95%CI:0.71-0.88) while the 5 mg
                                       on




                                                                              patients with thrombus resolution and thrombus persist-
                    twice was not (HR:0.88,95%CI:0.76-1.02). Major
                                                                              ence, no significative difference was noted about under-
                    bleedings were associated with both 2,5 mg twice
                                                                              lying cardiomyopathy (i.e., acute myocardial infarction,
                    (HR:1.58,95%CI:1.32-1.89) and 5 mg twice
                                  N




                                                                              primitive dilated cardiomyopathy), comorbidities (i.e.
                    (HR:1.4,95%CI:1.20-1.86). The NNT to prevent one
                                                                              atrial fibrillation, diabetes mellitus, smoking history,
                    CVE for LDR 2,5 mg bid was 66(49-122) and the NNH
                                                                              prior acute ischemic stroke or peripheral arterial dis-
                    to prevent major bleeding was 114(-375-751).
                                                                              ease), and treatment characteristics (i.e., median TTR
                    Conclusions: LDR 2,5 mg bid has a favourable risk-
                                                                              and concomitant dual antiplatelet therapy). While no
                    benefit profile in patients with CAD/PAD. A careful
                    selection of patients to treat with LDR is needed to      patients developed on-treatment acute ischemic stroke,
                    reduce the risk of major bleeding.                        acute peripheral embolism, and acute myocardial
                                                                              infarction, two patients (9.5%) developed an on-treat-
                                                                              ment major gastrointestinal bleeding (one in the upper
                                                                              and one in the lower gastrointestinal tract) and one
                      PO106
                                                                              patient (4.8%) developed an on-treatment genitourinary
                                                                              clinically relevant non-major bleeding. All bleeding
                    CLINICAL HISTORY OF PATIENTS WITH LEFT                    events occurred within the first three months of therapy.
                    VENTRICULAR THROMBOSIS RECEIVING                          Conclusions: Despite suboptimal TTR values, the
                    VITAMIN K ANTAGONISTS                                     cumulative incidence of thrombus resolution during the
                                                                              three recommended months of VKA therapy was
                    E. Valeriani1, T. Bucci2, F. Baratta3, D. Menichelli3,    roughly 40% and rose to roughly 60% if anticoagulant
                    F. Del Sole3, D. Pastori4, P. Pignatelli3                 therapy lasted for 12-months. Furthermore, AVK thera-




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                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    py protected patients against cardioembolic events and     probability to be the responsible lesion of GIB, using a
                    is associated with an acceptably low rate of major and     modified Saurin classification, by two independent gas-
                    clinically relevant non-major bleeding.                    troenterologists. Results: From November 2013 to
                                                                               March 2022, 208 patients (121 male), consecutively
                    Tabella 1.                                                 admitted to Santa Maria della Misericordia Hospital in
                                                                               Perugia for spontaneous DOACs-related GIBs were
                                                                               included. At entry, the mean patient age was 83±4 years
                                                                               (range 51 –95 years). 63 (30.1%%) patients presented
                                                                               GIB on dabigatran, 73 (35.1%) on rivaroxaban, 48
                                                                               (23.1%) on apixaban and 24 (11.5%) on edoxaban. In
                                                                               AF patients, median CHA2DS2VASC was 4.5 and
                                                                               median HAS-BLED was 2.6. According with ISTH cri-
                                                                               teria, GIB was classified as major in 73.1% of patients.
                                                                               At entry, mean Hb value was 8.8 gr per deciliter, mean
                                                                               creatinine value was 1.47 mg per deciliter. The site of
                                                                               bleeding was in upper GI tract in 79 patients (42.5%), in
                                                                               lower GI tract in 65 patients (34.9%), in both in 42
                                                                               patients (22.6%). In 10.6%, the endoscopic procedures
                                                                               were not performed or were not able to detect a source




                                                                                                       ly
                                                                               of bleeding. 58.7% of endoscopic lesions identified in




                                                                                               on
                                                                               upper gastrointestinal tract and 68.2% of endoscopic
                                                                               lesions identified in lower gastrointestinal tract were
                                                                               adjudicated as with high probability for being potential-
                                                                               ly bleeding lesions (of which 27.6% of cases with evi-




                                                                                          e
                                                                               dence of active bleeding or clot). A systemic reversal of
                                                                                   us
                                                                               anticoagulation was performed in 13.5% patients. The
                                                                               endoscopic procedures were performed urgently in
                                                                               52.0% of cases (of which 73.8% were in upper GI).
                                                                               al
                                                                               Fifty-four patients (85% of patients with evidence of
                                                                               active bleeding or clot) received an urgent local
                                                                        ci

                                                                               haemostasis. Conclusions: In this prospective cohort
                                                                               study, the upper GIBs were more frequent. In about 65%
                                                                  er


                                                                               of cases endoscopic procedures were able to identify
                                                                               lesions with high probability to be responsible of GIB
                                                             m



                                                                               and 25.9% of patients receiving an urgent local
                                                                               haemostasis.
                                                 om




                    Figura 1.
                                                                               PO108
                                              -c




                    PO107                                                      FCSAPP: A STRATEGIC NETWORK FOR A CLOSE
                                      on




                                                                               CONNECTION BETWEEN ANTICOAGULATION
                    ENDOSCOPIC FINDINGS OF PATIENTS WITH                       CLINICS
                                  N




                    DOACS-ASSOCIATED GASTROINTESTINAL
                    BLEEDING: RESULTS FROM PROSPECTIVE                         E. Antonucci1, S. Testa2, D. Barcellona3, P. Bucciarelli4,
                    COHORT STUDY                                               A. Ciampa5, E. Grandone6,7, G. Malcangi8,
                                                                               G. Rescigno9, A. Squizzato10, V. Toschi11, D. Poli12
                    M. Verso1, O. Morelli2, D. Castellani2, G. Bassotti2,
                                                                               1Fondazione
                    A. Vinci1, C. Becattini1                                                 Arianna Anticoagulazione, Bologna;
                                                                               2Dipartimento   di Medicina di Laboratorio, Centro
                    1Emergency     and Vascular Medicine-Stroke Unit,          Emostasi e trombosi, ASST Cremona, Cremona;
                                                                               3Dipartimento di Scienze Mediche e Sanità Pubblica,
                    University of Perugia; 2Gastroenterology Division,
                    University of Perugia, Italy                               Unità Semplice Dipartimentale di Emostasi e Trombosi
                                                                               Azienda Ospedaliero-Universitaria di Cagliari, Cagliari;
                                                                               4Centro Emofilia e Trombosi “Angelo Bianchi Bonomi”
                    Background and Aims: Gastrointestinal bleedings
                    (GIBs) are clinically relevant complications in patients   UOC Medicina Generale Emostasi e Trombosi
                    receiving long-term therapy with DOACs. The aim of         Fondazione IRCCS Ca’ Granda Ospedale Maggiore
                    the study was to describe the endoscopic characteristics   Policlinico, Milano; 5UOSD Centro Emostasi AORN
                    of DOACs-related GIB. Methods: A prospective cohort        SGMoscati, Avellino; 6IRCCS Casa Sollievo della
                    study of patients with DOACs-related GIBs was per-         Sofferenza, Poliambulatorio Giovanni Paolo II, Viale
                    formed. All endoscopic lesions were adjudicated for the    Padre Pio, San Giovanni Rotondo (FG); 7Dipartimento




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 152



         Poster




                    di Scienze Mediche e chirurgiche, Università degli Studi       Department of Hematology and Rare Diseases, Azienda
                    di Foggia, Foggia; 8Unità Operativa Semplice a Valenza         Ospedaliera Ospedali Riuniti Villa Sofia-Cervello,
                    Dipartimentale Centro Emofilia e Trombosi Azienda              Palermo, Italy
                    Ospedaliero-Universitaria Policlinico di Bari, Bari;
                    9Centro di Emostasi e Trombosi per la Diagnosi e la Cura       Patients (PTs) with β-thalassemia (β-T) show a chronic
                    delle Malattie Tromboemboliche ed Emorragiche DEA              hypercoagulability state due to numerous factors
                    PP.OO“Umberto I-A. Tortora-M. Scarlato”, ASL Salerno,          including iron overload, splenectomy and hemolysis. 1
                    Nocera Inferiore (SA); 10Centro di Ricerca Malattie            The increased thrombotic risk requires often the use of
                    Tromboemboliche e Terapie Antitrombotiche Università           antiplatelet and/or anticoagulant agents as primary
                    dell’Insubria, ASST Lariana, Ospedale Sant’Anna, San           and/or secondary prophylaxis. Despite the extensive
                    Fermo della Battaglia (Como); 11SIMT e Centro Trombosi         use of direct oral anticoagulants (DOACs) during the
                    ASST Santi Paolo e Carlo, Milano; 12SOD Malattie               past years, there is limited experience regarding the use
                    Aterotrombotiche, Azienda Ospedaliero Universitaria-           of DOAC in PTs with β-T. The studies on DOACs effi-
                    Careggi, Firenze, Italy                                        cacy and safety have not been conducted in PTs with β-
                                                                                   T, however, they may represent a valuable alternative,
                    Background and Aims: In the recent decades, the wide-          given the reduced half-life, decreased bleeding risk and
                    spread adoption of smartphones and digital innovations,        incidence of venous thromboembolism (VTE) recur-
                    has greatly changed everyday life and consumer behav-          rences and/or stroke respect to Vitamin K antagonist
                    ior in many ways. In addition, the widespread internet         (VKA). Only one study tested the effectiveness of




                                                                                                          ly
                    connectivity led to a significant change in human-tech-        rivaroxaban in a very small group of PTs with β-T and
                                                                                   non-valvular atrial fibrillation (NVAF).2 We herein




                                                                                                   on
                    nology interaction. Mobile health apps and medical apps
                    are becoming increasingly popular in a wide range of           present our experience with the use of DOACs in 17
                    health-related applications in almost all sectors of health-   PTs with β-T: 9 with NVAF and 8 with history of deep
                    care. The purpose of FCSApp is to provide to physicians        vein thrombosis and pulmonary embolism. The charac-




                                                                                              e
                    who referred to Italian Federation of Centers for the          teristics of PTs are reported in Table 1.
                    Diagnosis of Thrombotic Disorders and the Surveillance
                    of the Antithrombotic Therapies (FCSA) a tool to active-
                    ly share clinical suggestions, comments, scientific infor-
                                                                                       us
                                                                                   Table 1.
                                                                                   al
                    mation helpful for everyday practice. Methods: FCSApp
                    is developed by Prospero Multilab and is active from
                                                                           ci

                    October 2021. The app is available for iOS and Android,
                    it is free, and it works as an interactive chat. To access
                                                                     er


                    each physician member of FCSA must be register and
                    create an account; after login each participant can follow
                                                               m



                    and actively participate to the “discussion channels” that
                    meet his/her field of interest. In FCSApp there is a sec-
                                                  om




                    tion dedicated to news and communications from the sec-
                    retary office. Three Institutional channel (FCSA Board,
                    Regional FCSA Representative and FCSApp board)
                                               -c




                    were created to allow a fast communication among mem-
                    bers of each group. Results: From October 2021 about
                                       on




                    200 physicians have signed up; 10 channels have been
                    created. More than 30 clinical cases have been discussed
                    among users. All cases presented in the chat reported spe-
                                  N




                    cific clinical situation on patients generally excluded
                    from clinical trial; 20 articles on hemostasis and throm-
                    bosis have been shared. Conclusions: FCSApp is a
                    social networking app aimed to facilitate interaction,
                    research and collaboration between FCSA members.
                    Users demonstrate to be interested mainly in the discus-
                    sion of the clinical problems from their own practice.


                      PO109

                    EMPLOYMENT OF DIRECT ORAL
                    ANTICOAGULANTS IN Β-THALASSEMIA PATIENTS

                    S. Raso, L. Pitrolo, G. Calvaruso, A. Giangreco,
                    R. Di Maggio, D. Renda, A. Vitrano, G. Carmina,
                    A. Maggio




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Data was obtained from review of electronic health            controls during the follow-up of Rivaroxaban (p>0.05)
                    records. The median time of treatment was 36 months           while those of Apixaban were not different at trough
                    (IQR, 16-51) and during the follow-up 3 (18%) PTs             (p>0.05) but all lower than controls at peak (p=0,019).
                    developed superficial vein thrombosis and 1 (5%) intes-       Finally, DD levels during dabigatran treatment were
                    tinal ischemia while none experienced major bleeding,         lower than controls both at trough and peak (p=0.004
                    including PTs treated also with antiplatelet drug.            and 0.0016 respectively). Conclusions: Plasma levels
                    Thrombosis and thromboembolic events were treated             of DOAC are stable over time. No signs of hypercoag-
                    with low-molecular-weight-heparin (LMWH). Three of            ulability were found during the entire follow-up. These
                    4 PTs showed increased platelet count at the moment of        findings are reassuring in the long-term treatment with
                    thrombotic episode. None of PTs with thrombotic com-          these drugs.
                    plications was switched to another DOAC. Seven out of
                    17 PTs started DOACs as frontline anticoagulant treat-        Table 1.
                    ment and 10/17 PTs were shifted from a previous VKA
                    treatment. Rivaroxaban was the drug most often used
                    while dabigatran was unemployed. Frequently PTs with
                    β-T present liver and kidney damage, due both to course
                    of the disease and to hemosiderosis. Therefore, using
                    DOACs could be challenging, as they have been report-
                    ed to cause hepatotoxicity. However, in our experience




                                                                                                         ly
                    there were no adverse events reported and no need of




                                                                                                  on
                    dose modification in any PT. In conclusions, in our
                    experience anticoagulant therapy with DOACs in PTs
                    with βT is well tolerated, apparently safe, without recur-
                    rences of thrombotic events and significant bleeding




                                                                                             e
                    complications. Further studies are therefore needed in
                    order to assess the role of DOACs in PTs with β-T.
                                                                                      us
                                                                                  al
                    PO110
                                                                          ci

                    STABILITY OVER TIME OF DIRECT ORAL
                                                                    er


                    ANTICOAGULANTS
                                                              m



                    D. Barcellona1, M.F. Ruberto1, A. Mameli2, P. Schirru2,
                    S. Cornacchini2, F. Marongiu1
                                                  om




                    1Department   of Medical Sciences and Public Health,
                    University of Cagliari; 2Azienda Ospedaliero-
                    Universitaria di Cagliari, Italy
                                               -c




                    Background and Aims: Direct Oral Anticoagulants
                                       on




                    (DOAC) are the preferred drugs for treatment and pro-
                    phylaxis of venous thromboembolism and atrial fibril-
                                  N




                    lation. It is known that a high variability does exist. The
                    aim was to investigate whether the DOAC plasma lev-
                    els were stable during a long follow up (six months). D-
                                                                                  PO111
                    Dimer (DD) was also detected as an indicator of hyper-
                    coagulability. Methods: A total of 126 patients treated
                    as follows: 52 with Rivaroxaban 20 mg/qd, 53 with             PROPROTEIN CONVERTASE SUBTILISIN/KEXIN
                    Apixaban 5 mg/bid and 21 with Dabigatran 150 mg/bid.          TYPE 9 (PCSK9) IN THROMBOTIC PRIMARY
                    Dosages of DOAC were carried out at trough and after          ANTIPHOSPHOLIPID SYNDROME. THE
                    2h (peak). Anti Xa and direct thrombin inhibitor              MULTICENTER ATHERO-APS STUDY
                    (Hemosil Werfen) assays were used for measuring
                    drugs plasma levels. DD was detected by HS HemosIL            T. Bucci1, P.R.J. Ames2,3, V. Cammisotto4,
                    (Werfen). All dosages were carried out at the first, third    S. Bartimoccia4, M. Triggiani5, R. Parente5, A. Ciampa6,
                    and sixth month of treatment. Healthy controls were 55.       P. Pignatelli4, R. Carnevale7, D. Pastori4
                    Results: ANOVA for repeated measures showed no sta-
                    tistical differences among the three different DOAC           1Department    of General Surgery and Surgical
                    plasma levels both at trough and peak (Table 1).              Specialties “Paride Stefanini”, Sapienza University of
                    ANOVA (Kruskal-Wallis) sowed that DD levels (trough           Rome, Rome, Italy; 2Immune Response and Vascular
                    and peak) were not different among the dosages and            Disease Unit, CEDOC, Nova University Lisbon, Rua




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 154



         Poster




                    Camara Pestana, Lisbon, Portugal; 3Department of            an indirect effect on platelet function since they inter-
                    Haematology, Dumfries Royal Infirmary, Cargenbridge,        fere with the generation or activity of thrombin, but the
                    Dumfries, UK; 4Department of Clinical, Internal,            impact of Direct Oral Anticoagulants (DOACs) is large-
                    Anesthesiological and Cardiovascular Sciences,              ly unknown. Previous studies conducted with
                    Sapienza University of Rome, Rome, Italy; 5Department       Dabigatran, Apixaban and Rivaroxaban showed a
                    of Internal Medicine, Division of Allergy and Clinical      reduction in endogenous thrombin potential (ETP) and
                    Immunology, University of Salerno, Italy; 6Centro           in platelet aggregation induced by thrombin and tissue-
                    Emostasi A.O.R.N“SG Moscati”, Avellino, Italy;              factor (TF) in a dose dependent manner. Aim of this
                    7Department of Medical-Surgical Sciences and                study was to evaluate the effects of Edoxaban on
                    Biotechnologies, Sapienza University of Rome, Latina,       platelet function by in vitro and ex vivo studies.
                    Italy                                                       Methods: We evaluated platelet aggregation (PA),
                                                                                thrombin generation (TG) and thromboxane B2 (TXB2)
                    Background:          The      proprotein      convertase    levels in 20 healthy donors: samples were incubated in
                    subtilisin/kexin type 9 (PCSK9) is a novel cardiovascu-     vitro with increasing concentrations of Edoxaban [E50,
                    lar risk factor. Levels of PCSK9 in thrombotic primary      E150, E250 (ng/mL)] or vehicle as control. We also
                    antiphospholipid syndrome (PAPS) have never been            investigated the same parameters in 20 patients with
                    investigated. Methods: Cross sectional comparison of        Atrial fibrillation (AF) treated with Edoxaban (ex vivo
                    baseline characteristics of 91 PAPS patients enrolled in    study). A PAP-8 aggregometer was used to assess PA on
                    the multicenter ATHERO-APS cohort study. PCSK9              PRP samples, induced by ADP (5 μM), TRAP-6 (10




                                                                                                       ly
                    levels were categorized into tertiles and the association   μM), Human Thrombin (THR, 0.182 mU/μL) and TF.
                                                                                TG was measured (both in the presence and absence of




                                                                                                on
                    with arterial and recurrent thrombosis were assessed by
                    univariable and multivariable regression analysis.          thrombomodulin-TM) using the Calibrated Automated
                    Results: Median age was 51 years and 71.4% (n=65)           Thrombogram System (CAT). Serum TXB2 was meas-
                    were women. Overall, 33% (n=30) experienced an arte-        ured by using the TXB2 EIA kit, according to the man-




                                                                                          e
                    rial event while 31% (n=28) had recurrent thrombotic        ufacturer’s instructions. Results: The incubation of
                    events. Median PCSK9 levels were 1243 (1100-1650)
                    pg/ml. Patients in the third PCSK9 tertile (>1458 pg/ml)
                    showed a higher prevalence of dyslipidemia, lupus anti-
                                                                                   us
                                                                                PRP with different Edoxaban concentrations signifi-
                                                                                cantly reduced TF-induced PA with respect to vehicle
                                                                                [E50 by 21% (p=0.033), E150 by 33% (p=0.004), and
                                                                                al
                    coagulant positivity and a history of previous arterial     E250 by 52% (p<0.001)]. TF-induced PA was signifi-
                    and recurrent thrombosis than patients in the first and     cantly lower in patients treated with Edoxaban than in
                                                                        ci

                    second tertile. PCSK9 levels were higher in arterial than   controls (p<0.001). ADP and TRAP-6-induced PA was
                    venous thrombosis (1502 vs. 1180 pg/ml, p=0.002), and       not inhibited by any Edoxaban concentrations in the in
                                                                  er


                    in patients with recurrent vs isolated thrombosis (1680     vitro study, and also ex vivo experiments failed to
                    vs. 1150 pg/m, p<0.001). High plasma PCSK9 levels           demonstrate any difference between ADP and TRAP-6-
                                                             m



                    were associated with a 4-fold increase risk for arterial    induced PA from AF patients treated with Edoxaban and
                    events and with a 10-fold increase risk for recurrent       controls. THR-induced aggregation in E150 group
                                                 om




                    thrombosis after adjustment for confounding factors.        showed a trend towards a reduction, though did not
                    Conclusions: These preliminary data suggest that in         reach the statistical significance. Among the parameters
                    PAPS, PCSK9 levels are associated with arterial and         related to TG, Lag Time (-TM) was significantly
                                              -c




                    recurrent thrombosis. Its role as a possible therapeutic    (p<0.001) and positively related to Edoxaban concen-
                    target in PAPS needs further studies.                       trations. Patients showed more prolonged Lag Time
                                      on




                                                                                values (p=0.031) with respect to those observed in con-
                                                                                trols. ETP and Peak (-TM) were significantly reduced
                      PO112                                                     in vitro (p<0.001) by the incubation of Edoxaban in a
                                  N




                                                                                dose dependent manner. In in vitro study, ETP ratio val-
                                                                                ues were significantly reduced according to increasing
                    CHARACTERIZATION OF EDOXABAN EFFECTS ON                     Edoxaban concentrations. AF patients showed reduced
                    PLATELET FUNCTION                                           levels of ETP ratio with respect to controls (p<0.001).
                                                                                We found a 24% decrease in serum TXB2 concentration
                    G. Barbieri1, A.M. Gori2, F. Cesari3, E. Sticchi2,          in the E250 group vs control (p<0.01), while the reduc-
                    A. Rogolino3, R. Orsi1, A. Bertelli1, R. De Caterina4,      tion is not significant in the other Edoxaban concentra-
                    B. Giusti2, R. Marcucci2                                    tions (Figure 1). Conclusions: Our data show that
                                                                                Edoxaban is able to interfere with platelet function. In
                    1University of Florence, Department of Experimental         particular, it significantly reduces TF-induced platelet
                    and Clinical Medicine, Florence; 2University of             aggregation in a dose-dependent manner and also TG,
                    Florence, Department of Experimental and Clinical           although thrombin-induced aggregation is not affected
                    Medicine, Atherothrombotic Diseases Center, Careggi         by the drug, supporting Edoxaban’s indirect effect on
                    Hospital, Florence; 3Atherothrombotic Diseases Center,      thrombin by inhibiting FXa. In addition, the reduction
                    Careggi Hospital, Florence; 4Cardiology division,           of TXB2 levels by Edoxaban suggests that the drug is
                    University of Pisa, Italy                                   endowed with an antiplatelet effect, which may, in turn,
                                                                                lead to the delayed/reduced formation of coagulation
                    Background: All anticoagulants are expected to have         complexes reinforcing its antithrombotic potential.




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                                                                                 in premature CAD. Methods: We retrospectively eval-
                                                                                 uated data form patients admitted for acute coronary
                                                                                 syndrome at Department of Cardiology of Careggi
                                                                                 University Hospital, Florence, between 1st January
                                                                                 2021 and 28th February 2022. Inclusion criteria were:
                                                                                 age <50 years for men and <55 years for women; con-
                                                                                 firmed diagnosis of acute coronary syndrome at hospi-
                                                                                 tal discharge. Results: We selected 78 patients (12 F
                                                                                 and 66 M; median age: 46 years). LDL median values
                                                                                 at admission were 121.87 mg/dl). Four out of 78
                                                                                 patients (5.1%) had LDL levels higher than 190 mg/dl,
                                                                                 suggesting the diagnosis of Heterozygous Familial
                                                                                 Hypercholesterolemia (with respect to the expected
                                                                                 prevalence in the general population: 1/250 (0.004%).
                                                                                 No patient had hepatic disease or thyroid disease.
                                                                                 Twenty out of 78 patients (25.6%) had triglycerides lev-
                                                                                 els higher than 150 mg/dl; 17/78 (21.7%) with levels
                                                                                 higher than 200 mg/dl. As regards Lp(a) levels, a
                                                                                 dosage was performed in only 34/78 patients (43.5%).




                                                                                                         ly
                                                                                 Among these, elevated Lp(a) levels were documented




                                                                                                 on
                                                                                 in 11/34 (32.4%). In particular, 4/11 (11.8%) had
                                                                                 extreme elevated Lp(a) levels (i.e. higher than 800
                                                                                 mg/L) (Figure 1). In a patient we documented both
                                                                                 extreme Lp(a) levels and LDL levels higher than 190




                                                                                             e
                                                                                 mg/dl; in the other patients elevated Lp(a) levels were
                                                                                     us
                                                                                 the only lipid parameter out of range, and the only risk
                                                                                 factor for premature CAD. At a multivariate analysis
                                                                                 adjusted for classical risk factors, elevated Lp(a) levels
                                                                                 al
                                                                                 were significantly associated with the risk of premature
                                                                                 CAD (OR: 2.4 95%CI 1.9-3.6); p<0.001).
                                                                         ci

                                                                                 Conclusions: these results demonstrated that: 1) it is
                                                                                 urgent, from a clinical point of view, to apply screening
                                                                   er


                                                                                 protocols in premature CAD including Lp(a) dosage; 2)
                                                                                 elevated Lp(a) levels, and in particular extreme elevat-
                                                             m



                    Figure 1.                                                    ed Lp(a) levels, have an elevated prevalence in a popu-
                                                                                 lation of premature CAD and are significantly and inde-
                                                 om




                                                                                 pendently associated with the disease.
                    PO113
                                              -c




                    HYPERLIPOPROTEIN(A) AND PREMATURE
                                       on




                    CORONARY ARTERY DISEASE: A CALL FOR
                    ACTION
                                  N




                    T. Capobianco, W. Iannotti, A. Sammartino, M. Casini,
                    S. Tedesco, F. Brogni, E. Lotti, M. Berteotti, R. Valenti,
                    A. Migliorini, N. Marchionni, C. Di Mario, R. Marcucci

                    Dipartimento    Cardiotoracovascolare,        Azienda
                    Ospedaliero-Universitaria Careggi, Firenze, Italy
                                                                                 Figure 1.
                    Background and Aims: Elevated Lipoprotein (a) lev-
                    els are well recognized as a genetic risk factor for
                    atherothrombotic events. Recently, new therapeutic           PO114
                    approaches – such as small interfering RNA (siRNA)
                    and antisense oligonucleotides (ASOs) – have been
                    developed and are under evaluation in phase 3 trials.        PREDICTORS OF ISCHEMIC AND HEMORRHAGIC
                    Evidences from literature suggest the need for screen-       EVENTS IN PATIENTS WITH ATRIAL FIBRILLATION
                    ing Lp(a) in premature coronary artery disease. Aim of       UNDERGOING PERCUTANEOUS CORONARY
                    our study was to evaluate, in a real world scenario: 1)      INTERVENTION
                    the percentage of premature CAD in whom clinicians
                    choose to screen Lp(a); 2) the prevalence of hyperLp(a)      M. Berteotti1, A.M. Gori1, A. Bertelli1, B. Bandinelli1,




                                                              [Hematology Reports 2021; 13:s3]                                            [page 155]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 156



         Poster




                    G. Grossi2, N. Ciardetti2, B. Giusti1, G. Barbieri1,          p=0.001), peripheral artery disease (HR 3.17, 95% C.I.
                    E. Lotti1, R. Valenti2, C. Di Mario1, N. Marchionni1,         1.06-9.43, p=0.038), white blood cells count (HR 1.89,
                    R. Marcucci1                                                  95% C.I. 1.14-3.12, p=0.014), eGRF (HR 0.41, 95%
                                                                                  C.I. 0.22-0-76, p=0.005) and D-dimer (HR 1.64, 95%
                    1Department  of Clinical and Experimental Medicine,           C.I. 1.05-2.58, p=0.031). Conclusions: In a real-world
                    University of Florence, Florence, Italy; 2Division of         unselected population, a significant incidence of
                    Interventional Cardiology, Cardiothoracovascular              ischemic as well as hemorrhagic events was observed
                    Department, Careggi University Hospital, Florence,            both in patients on TAT and DAT early after discharge.
                    Italy                                                         Our data documented the relevance of the entity of
                                                                                  platelet inhibition in patients with ACS suggesting that
                    Background and Aims: Antithrombotic management                a laboratory evaluation of drug response might be use-
                    of patients with atrial fibrillation (AF) undergoing per-     ful in personalizing antithrombotic therapy in this set-
                    cutaneous coronary intervention (PCI) can be challeng-        ting. Besides D-Dimer resulted an independent predic-
                    ing. Recent direct oral anticoagulant (DOAC) trials           tor of both ischemic and bleeding events, as well as
                    demonstrated the safety of double antithrombotic thera-       death.
                    py (DAT) in comparison with triple antithrombotic ther-
                    apy (TAT) in this setting. However, this benefit is coun-
                    terbalanced by a higher risk of ischemic, mainly stent-       PO115
                    related, events. We therefore sought to identify possible




                                                                                                         ly
                    laboratory predictors of bleeding and ischemic risk in a
                    real-world population with concomitant AF and PCI.            EXTRACELLULAR VESICLES NUMBER AND CELL




                                                                                                  on
                    Methods: All consecutive patients with history of AF          SUBTYPE MAY BE INFLUENCED BY DIABETES
                    discharged from our cardiology ward with DAT or TAT           MELLITUS AND METFORMIN IN PATIENTS AT
                    after a PCI from April 2018 to March 2021 were                HIGH CARDIOVASCULAR RISK




                                                                                            e
                    enrolled in an observational registry. For all subjects,
                                                                                  P.G. Simeone1, R. Liani1, G. Bologna1, R. Tripaldi1,
                    blood serum samples were collected and tested for D-
                    Dimer, thrombin generation time, clot lysis time,
                    platelet reactivity by arachidonic acid and ADP, ery-
                                                                                     us
                                                                                  A. Di Castelnuovo2, P. Simeone1, D. D’Ardes1,
                                                                                  S. Miscia1, F. Cipollone1, M. Marchisio1, A. Consoli1,
                                                                                  P. Lanuti1, F. Santilli1
                                                                                  al
                    throcyte aggregation and deformability, oxidative stress
                    and DOAC concentration if appropriate. Major adverse
                                                                                  1Department of Medicine and Aging, Center for
                                                                          ci

                    cardiac and cerebrovascular events (MACCE) and
                    major hemorrhagic or clinically relevant non major            Advanced Studies and Technology (CAST), Chieti, Italy;
                                                                    er


                                                                                  2Mediterranea Cardiocentro, Napoli, Italy
                    bleeding events together with therapeutic changes were
                    recorded for all patients. Results: A total of 147 patients
                                                              m



                    were included (70.1% after acute coronary syndromes);         Background and Aims: Extracellular vesicles (EV)
                    the mean age was 78±8 years and 48 (33%) were                 represent a population of small vesicles deriving from
                                                  om




                    women. Ninety-one patients (62%) were discharged              all types of cells, generated by the extroflection of the
                    with TAT. Both in TAT and in DAT group DOACs were             plasma membrane, and released into the circulation. EV
                    preferred (58% and 77%, respectively). In 93.4% of            can have both pro- and anti-atherothrombotic effects
                                               -c




                    patients, clopidogrel was chosen as P2Y12 inhibitor.          depending on the clinical setting, origin cell, stimuli,
                    The median follow-up was 401 days (IQR 241-588).              and different treatments might affect their levels. The
                                       on




                    MACCE occurred in 26 cases (17.7%), while hemor-              primary endpoint of our study was to compare the
                    rhagic events in 25 (17%), 14 of which were major             amount of circulating EV and specific EV subtypes
                    (9.5%), with no significant differences between TAT           derived from platelets, endothelial cells, and leucocytes
                                  N




                    and DAT group. The incidence of all-cause death and           in subject at high CV risk, with and without T2DM, and
                    cardiovascular death was 12.9% and 7.4%, respectively.        if any ongoing anti-diabetic drugs could affect the lev-
                    The independent predictors of MACCE at COX regres-            els of EV. Methods: Fifty-nine T2DM patients and
                    sion analysis were acute coronary syndrome presenta-          forty patients without T2DM with or without prior vas-
                    tion (HR 8.70, 95% C.I. 1.16 to 65.42, p=0.036),              cular disease, were enrolled, this was a cross-sectional
                    P2Y12 dependent platelet reactivity-PRU (HR 1.65,             study, extracellular vesicles were evaluated by flow
                    95% C.I. 1.15 to 2.38, P=0.007) and D-Dimer (HR               cytometry analysis. Results: After propensity score
                    1.34, 95% CI 1.03 to 1.75, p=0.030). Platelet function        matching the levels of total EV (Cohen d=0.55,
                    estimated by PRU, was significantly affected by the *2        p=0.064), total Annexin+ EV (d=0.49, p=0.043), EV
                    genotype: 155.5 (102-218.7) in wild-type vs 204 (96-          CD41a+ platelet derived EV (d=0.51, p=0.014),
                    246) in carriers of *2 allele (p<0.05). The independent       CD41a+/AnnexinV+ platelet derived EV (d=0.50,
                    predictors of bleeding were active smoke (HR 3.31,            p=0.019) and CD45 leukocyte derived EV (d=0.19,
                    95% C.I. 1.04-10.34, p=0.043), previous myocardial            p=0.012) were lower in diabetic patients vs. those with-
                    infarction (HR 0.27, 95%C.I. 0.08-0.96, p=0.043), left        out T2DM. Interestingly, platelet-derived EV positive
                    main disease (HR 2.76, 95% C.I. 1.07-7.09, p=0.035)           for AnnexinV were lower in patients on treatment with
                    and D-dimer (HR 1.63, 95% C.I. 1.12-2.37, p=0.011).           metformin (d=0.72, p=0.012) (Figure 1). Conclusions:
                    Independent predictors of all-cause mortality were the        Our study showed that, among high CV risk patients,
                    following: male gender (HR 0.13, 95% CI 0.04-0.41,            treated with the state-of-the-art preventive strategies,




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    T2DM is associated with lower levels of total, platelet-     uals was analyzed (55% M and 45% females). As
                    and leucocyte-derived EV, and that this difference may       regards subjects on lipid-lowering therapies, we found
                    be accounted for, at least in part, by ongoing treatment     that female gender was associated with a significantly
                    with metformin, the most widely prescribed oral antidi-      lower probability to have LDL levels at target (OR:
                    abetic agent.                                                0.58-0.01; p<0.000) in patients with MACCE with or
                                                                                 without diabetes, in a model adjusted for age, district
                                                                                 area, and the presence of cardiovascular risk factors and
                                                                                 comorbilities (renal failure, heart failure, atrial fibrilla-
                                                                                 tion). This result was confirmed also in the analysis
                                                                                 conducted in subjects without lipid-lowering therapies
                                                                                 (OR: 0.56-0.01; p<0.000). No significant differences
                                                                                 were documented by stratifying for the presence of clin-
                                                                                 ical pathways aimed to the active call of the patients by
                                                                                 clinicians (“Initiative Health”). Discussion: These
                                                                                 results demonstrated that females have a significantly
                    Figure 1.                                                    lower probability to have LDL cholesterol levels at tar-
                                                                                 gets suggested by guidelines. This datum is confirmed
                                                                                 even in the absence of lipid-lowering therapies. These
                    PO116                                                        results call for action aimed to: 1) education for general




                                                                                                          ly
                                                                                 population and for patients with MACCE with a specif-




                                                                                                  on
                    LDL TARGET LEVELS IN MACCE PATIENTS: AN                      ic target on females; 2) information for clinicians for
                                                                                 the need of a lipid-lowering therapy gender-tailored; 3)
                    (UN)EXPECTED GENDER BIAS. A SNAPSHOT
                                                                                 organization of clinical pathways - from admission to
                    FROM TUSCANY                                                 recovery and follow-up - with a specific target on lipid




                                                                                            e
                                                                                 lowering therapies ad gender.
                    E. Lotti1, P. Francesconi2, E. Mannucci1, A. Zuppiroli1,
                    G. Casolo1, D. Grisillo1, N. Ferrante3, F. Profili2,
                    R. Marcucci1
                                                                                     us
                                                                                 PO117
                                                                                 al
                    1Department   of Experimental and Clinical Medicine,
                                                                                 THE ASSOCIATION BETWEEN PAD AND
                                                                         ci

                    University of Florence; Atherothrombotic center, AOU
                    Careggi, Itay; 2ARS-Agenzia regionale di sanità della        DIABETES: PRELIMINARY FINDINGS FROM A
                                                                   er


                    Toscana, Firenze, Italy; 3Novartis FarmaVarese, Italy        PROSPECTIVE STUDY
                                                             m



                    Background: Collection of epidemiological data and           A. Abenante1, M. Dalle Fratte1, R. Corso1,
                    information on related healthcare is crucial for monitor-    C. Dedionigi1, G. Fabro1, C. Fumagalli1, I. Giarretta2,
                                                 om




                    ing healthcare appropriateness and the fulfillment of        R. Landi3, S. Marino1, D. Mastroiacovo2, M.C. Naim1,
                    clinical needs. Limited data are available on the preva-     E. Nicolini2, P. Pagani1, A. Bonaventura2, F. Dentali1
                    lence of LDL cholesterol in Italian population, in partic-
                    ular in the very high risk population of MACCE               1Università degli Studi dell’Insubria di Varese; 2UO di
                                              -c




                    patients. Aims: This project aimed to estimate how           Medicina Interna, Ospedale di Circolo di Varese;
                    many patients had LDL cholesterol at the target levels       3Università degli Studi del Piemonte Orientale, Novara,
                                       on




                    suggested by the international guidelines by analyzing       Italy
                    regional administrative data. Methods: By linking
                                  N




                    health administrative and laboratory data, we collected      Background and Aims: Peripheral artery disease (PAD)
                    data from seven Local Health Districts in Tuscany on         refers to a partial or complete occlusion of the arterial
                    residents aged ≥45 years. Inclusion criteria were: 1) at     vessels of the lower limbs. Patients with diabetes mellitus
                    least one LDL measurement in the year; 2) a history of       (DM) have more than two-fold increased prevalence of
                    major adverse cardiac and cerebrovascular events             PAD compared with the general population. Despite its
                    (MACCE, defined as myocardial infarction and/or              epidemiological and clinical importance, PAD remains
                    stroke and/or revascularization of the coronary arteries,    largely underdiagnosed and undertreated. Therefore, this
                    the carotids, or peripheral arteries) and/or type 2 dia-     study investigated PAD in DM patients by measuring
                    betes (T2D). Cohorts were defined at 1st january 2019        ankle-brachial-index (ABI). Methods: Consecutive
                    and 1st january 2020. Lipid lowering therapy use             patients referring to the Diabetes Outpatient Clinics of
                    (defined as ≥2 prescription fillings or ≥75% coverage of     the ASST Settelaghi (Varese) were screened for PAD
                    treatment days) over the past 6 months if LDL choles-        since June 2018 and included in the database. Patients
                    terol (LDL-C) data were available or over the past year      with gestational DM or those with a history of type I DM
                    if no LDL-C data were available was monitored. The           shorter than 10 years were excluded. All patients signed
                    outcome was the number of patients with on-treatment         an informed consent. Every subject underwent a full clin-
                    LDL optimal levels, as defined by levels <55 mg/dl for       ical evaluation, ABI measurement and filled the San
                    pts with MACCE and <70 mg/dl for pts with diabetes           Diego Claudicatio questionnaire. A further diagnostic
                    without MACCE. Results: A cohort of 174,200 individ-         evaluation with Doppler ultrasound was planned for all




                                                              [Hematology Reports 2021; 13:s3]                                               [page 157]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 158



         Poster




                    patients with ABI <0.9 or for those with non-compress-          69 y/o woman is admitted into the emergency room com-
                    ible arteries. For the current analysis, patients with ABI      plaining of impaired movement of left arm and leg upon
                    <0.9 or those with a known diagnosis of PAD were con-           awakening. Upon arrival, she is alert, normotensive with
                    sidered as having PAD. Comparisons between patients             a rhythmic pulse and a left hemiparesis and hemianesthe-
                    were performed using Mann-Whitney U test or Fisher’s            sia is noted. She has a history of arterial hypertension,
                    exact test, as appropriate. All variables with p <0.10 were     diabetes mellitus, hypercholesterolemia and hypothy-
                    included in a backward stepwise logistic regression             roidism. Imaging shows subcortical ischemic outcomes
                    model to evaluate independent risk factors for PAD              in the left frontal lobe and right cerebellum and two bilat-
                    occurrence. Values were expressed as odds ratio (OR)            eral subacute thalamic lesions, the most recent on the
                    with 95% confidence interval (CI). A two-sided p<0.05           right. The patient is not a candidate for acute phase ther-
                    value was considered as statistically significant.              apy and therefore acetylsalicylic acid 300 mg/die and
                    Statistical analyses were performed using IBM SPSS              hight dose statin is started. During hospitalization, she
                    Statistics for Mac version 27.0 (IBM Co., Armonk, NY,           experiences chest pain radiating to the back. A contrast-
                    USA). Results: A total of 797 consecutive DM patients           enhanced CT scan shows multiple filling defects of the
                    were screened. Among them, 316 were women (39.6%).              pulmonary circulation, multiple splanchnic aneurysms
                    One hundred and seven (13.4%) patients were diagnosed           and dissection of the hepatic artery. In agreement with
                    with PAD, while 690 (86.6%) were not affected. The              the neurologist, anticoagulant therapy with LMWH is set
                    multivariate logistic regression analysis showed that age       up. The study of cerebral circulation reveals a VBD with
                    >65 years (OR 2.1; 95% CI, 1.25 to 3.62; p=0.005),              a maximum diameter of 6.2 mm. A coronary angiogra-




                                                                                                             ly
                    chronic kidney disease (OR 2.8; 95% CI, 1.30 to 3.31;           phy shows multiple aneurysms at this level as well. PET




                                                                                                     on
                    p=0.002), retinopathy (OR 2.39; 95% CI, 1.40 to 4.07;           with 18F-FDG is also considered to rule out vasculitis.
                    p=0.001) and heart failure (OR 1.80; 95% CI, 1.01 to            Transesophageal echocardiogram is performed, detecting
                    3.19; p=0.044) were significantly associated with PAD           patency of the foramen ovale. No signs of deep venous
                    diagnosis. Furthermore, carotid atherosclerosis showed a        thrombosis are found with doppler of the lower limbs.




                                                                                               e
                    positive association with PAD (OR 1.55; 95% CI, 0.98 to         Vascular surgeons recommended follow-up of splanch-
                    2.45; p=0.057). Conclusions: This preliminary analysis
                    of our ongoing study showed that several common
                    comorbidities are significantly associated with PAD.
                                                                                        us
                                                                                    nic aneurysms without any present surgical indication. A
                                                                                    diagnosis suspect Loeys-Dietz syndrome was made after
                                                                                    a full evaluation with an expert in collagen disease.
                                                                                    al
                    Moreover, the rate of undiagnosed PAD among DM                  Conclusions: The pulmonary embolism associated with
                    patients is not trivial. An early screening in this high-risk   cerebral ischemia in a patient with patent foramen ovale
                                                                            ci

                    subgroup of patients should allow easier diagnosis and          and coronary aneurysms, would lead to a long-term anti-
                    more effective treatment. Hence, a larger number of             coagulant therapy in secondary prevention, but VBD
                                                                     er


                    patients and further investigation is eagerly needed in the     increases complexity in this case. In this patients, the few
                    next future.                                                    data available show no evidence of superiority between
                                                               m



                                                                                    DOACs and VKAs in the prevention of ischemic events.
                                                                                    On the other hand it emerges that the greatest risk of
                                                   om




                      PO118                                                         bleeding complications is related to dilations greater than
                                                                                    6.5 mm or in case of a growth greater than 2 mm per year,
                                                                                    regardless the antithrombotic therapy. Therefore, the
                    VERTEBROBASILAR DOLICHOECTASIA IN THE
                                                -c




                                                                                    therapeutic choice with the highest safety profile seemed
                    CHOICE OF ANTICOAGULANT THERAPY:
                                                                                    to be Dabigatran 150 mg/db and a radiological follow-up
                                        on




                    A CASE REPORT                                                   to monitor the evolution of VBD. VBD is a variable to
                                                                                    consider when choosing antithrombotic therapy in
                    F. Crudele1,2, G. Guidotti1,2, E. Lotti1,2, D. Poli1,2,         patients with cerebrovascular events. In our opinion a
                                   N




                    L. Mannini1,2, C. Sarti3, F. Meucci1,4, R. Marcucci1,2          subgroup of the complications related to anticoagulant
                    1Department
                                                                                    therapy may be related to the presence of undiagnosed
                                 of Experimental and Clinical Medicine,             VBD. Further studies are needed to provide more valu-
                    University    of    Florence,     Florence,     Italy;          able data on this specific topic which may affect a dis-
                    2Atherothrombotic Center, AOU Careggi, Florence,
                                                                                    crete number of patient.
                    Italy; 3Department of NEUROFARBA, University of
                    Florence, Florence, Italy; 4Structural Interventional
                    Cardiology, AOU Careggi, Florence, Italy
                                                                                    PO119
                    Background: Dolichoectasia of the vertebro-basilar
                    artery (VBD) is defined by Smoker’s criteria as dilation        THROMBO-EMBOLIC RISK IN CARDIAC
                    greater than 4.5 mm, elongation and tortuosity of the           TRANSTHYRETIN AMYLOIDOSIS:
                    basilar artery. It is found in 9% of people over 60 years       A CASE REPORT
                    old. Etiology is unknown and it is associated with female
                    sex, arterial hypertension and diabetes mellitus. It is a       F. Crudele1, G. Guidotti1, A. Argirò2, E. Lotti1, D. Poli1,
                    possible cause of cerebrovascular events, both ischemic         L. Mannini1, F. Perfetto2, F. Cappelli2, R. Marcucci1
                    and hemorrhagic, and its mortality at 5 years after the
                    first clinical manifestation is about 30%. Case Report: A       1Department    of Experimental and Clinical medicine,




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 159



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    University of Florence; Atherothrombotic Center, AOU         ple of how complex it can be to manage these patients,
                    Careggi, Florence, Italy; 2Tuscan Regional Amyloidosis       taking into account the underlying disease and the fre-
                    Centre, AOU Careggi, Florence, Italy                         quent associated comorbidities. It is desirable that
                                                                                 more evidence be collected to refine the therapeutic
                    Introduction: Systemic amyloidosis is an extracellular       options in these patients. Furthermore, the routine
                    storage disease of insoluble protein fibrils. Its preva-     dosage of the concentrations of DOACs during adverse
                    lent cardiac involvement subtypes are due to the depo-       events provides useful information in the subsequent
                    sition of amyloid from immunoglobulin light chains or        management of the patient.
                    from transthyretin (ATTR). In recent years, ATTR
                    based amyloidosis incidence has had a marked increase
                    thanks to the greater diagnostic capacity: it is estimated   PO120
                    that up to 13% of subjects with heart failure with pre-
                    served ejection fraction may be affected by aTTR.
                                                                                 EVOLOCUMAB-RESISTANT
                    Other possible complications include arrhythmic
                    events, mostly atrial fibrillation (AF), and an increased    HYPERCHOLESTEROLEMIA IN A PATIENT WITH
                    risk of cardiogenic thrombo-embolic events. In these         PRIMARY BILIARY CHOLANGITIS.
                    patients intracavitary thrombi are more frequently           A CASE REPORT
                    detected at the time of an attempt at AF cardioversion,
                    despite adequate anticoagulation therapy. The                V. Liberati, E. Lotti, F. Crudele, E. Sticchi, B. Giusti,




                                                                                                          ly
                    increased thrombotic risk appears to be related to the       R. De Cairo, A. Rogolino, R. Marcucci
                    degree of amyloid infiltration and the severity of car-




                                                                                                  on
                    diac dysfunction. Case Report: In March 2022, we             Dipartimento medicina sperimentale e clinica,
                    evaluated Mr. F.S., aged 81, affected by transthyretin       Università di Firenze, Malattie Aterotrombotiche, AOU
                    cardiac amyloidosis from January 2020 and heart fail-        Careggi, Italy




                                                                                            e
                    ure with preserved ejection fraction and AF as compli-
                    cations. Patient’s comorbidities are stage III renal fail-
                    ure, as well as arterial hypertension, diabetes mellitus
                    and dyslipidemia. He is also suffering from non-critical
                                                                                     us
                                                                                 Background: Hypercholesterolemia is a common find-
                                                                                 ing in almost 75% of patients with primary biliary
                                                                                 cholangitis (PBC). Hypercholesterolemia is a well-
                                                                                 known modifiable risk factor for cardiovascular disease
                                                                                 al
                    carotidopathy in follow up. At the time of diagnosis,
                    warfarin therapy was initiated in the presence of a          in the general population, however in PBC’s patients
                                                                         ci

                    CHA2DS2-VASc score of 5. Subsequently, in February           the role of hypercholesterolemia in cardiovascular dis-
                    of the same year, he had an ischemic stroke during           ease remains controversial. Most of the studies had not
                                                                   er


                    well-conducted therapy (INR 3.01 at the time of the          found a statistically significant increase of atherosclero-
                    event). For this reason, warfarin was replaced with          sis incidence in patients with PBC and hypercholes-
                                                             m



                    Dabigatran 110 mg / bd, according to renal function.         terolemia compared with age and sex-matched controls.
                    He came to our attention for an episode of sudden            Among factors contributing to possible decrease athero-
                                                 om




                    blanching of the forearm and left hand, associated with      genesis in patients with hypercholesterolemia and PBC
                    intense pain and burning paresthesia, which occurred         there is LipoproteinX (LpX), an atypical lipoprotein
                    about 15 days earlier. At the time of the event, all the     rich in phospholipid and nonesterified cholesterol with
                                              -c




                    therapy was assumed regularly, including Dabigatran,         a similar density to LDL, found in cholestasis states
                    and any other associated symptoms or significant             (e.g. PBC) as a result of biliary lipids reflux into the
                                       on




                    events was not reported. He did not turn to the general      plasma. LpX seems to delay atherosclerosis process by
                    practitioner for spontaneous resolution of the event in      preventing LDL-oxydation and the adverse effects of
                    a few days. Subsequently, the reference cardiologist         oxidated-LDL on the endothelium; therefore it has been
                                  N




                    highlighted stability of the echocardiographic findings      hypothesized that hypercholesterolemia in PBC might
                    and good haemodynamic balance, interpreting the              even have a cardiovascular protective effect. LpX does
                    episode as a spontaneously resolved acute ischemia. At       not contain apolipoprotein B (ApoB), so it can’t be re-
                    the time of our evaluation, the patient complained of        captured by LDL receptor (LDLR); this could explain a
                    residual numbness in the hand, in the absence of clear       possible failure in reducing cholesterol concentration
                    signs of ischemia. An anti-IIa activity dosage was fully     with PCSK9 inhibitors in PBC’s patients. Case Report:
                    in the therapeutic range (with normal trough values of       This paper reports a case of hypercholesterolemia
                    140 ng/ml). Since it was fully in the therapeutic range,     refractory to the PCSK9 inhibitor Evolocumab in a 58
                    a therapy based on acetylsalicylic acid 75 mg/day was        years-old woman with a recent diagnosis of PBC. She
                    prescribed in addition to anticoagulation. Conclusions:      was admitted in our Centre for marked hypercholes-
                    Few data are available on patients with aTTR cardiac         terolemia. Lipid panel showed: total cholesterol 375
                    amyloidosis with recurrent ischemic events on antico-        mg/dl, LDL cholesterol 260 mg/dl and HDL cholesterol
                    agulation therapy. Data from literature and from ran-        81 mg/dl. She did not tolerate statins, fibrates, ezetim-
                    domized trials are not able to define the better             ibe and cholestyramine. Evolocumab wasn’t effective
                    antithrombotic strategy in this clinical setting.            on reducing LDL-C levels. Genetic analysis with NGS
                    Therefore we decided to increase the intensity of the        for familial hypercolesterolemia (FH) did not show
                    therapy adding an antiplatelet agent, because of multi-      genetic alterations of the PCSK9, ApoB and LDRLR
                    ple atherothrombotic risk factors. This case is an exam-     genes. In December 2021 she started complaining




                                                              [Hematology Reports 2021; 13:s3]                                               [page 159]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 160



         Poster




                    asthenia and itch. Blood tests showed an important          right frontal bone Ewing sarcoma, developed sympto-
                    increase of cholestasis indices (FA, GGT, bilirubin, bil-   matic right internal jugular vein occlusion (May 2021),
                    iary acids) and transaminases (AST, ALT). Anti-nuclear      thus enoxaparin 8000 UI twice/die was started fol-
                    antibody (ANA), anti-mitochondrial antibody (AMA)           lowed by CVC removal and reduction to 8000 UI/die
                    and anti-mitochondrial M2 antibody were positive.           after 1 month (weight 81 kg). In July chemotherapy
                    Fibroscan showed a fibrosis score of F3. The patient        was stopped for disease remission. A follow-up CT
                    was diagnosed with primary biliary cholangitis and a        scan in August showed occluding thrombosis of the
                    medical therapy with ursodeoxycholic acid (UDCA)            internal jugular, subclavian and anonymous right veins
                    was started. Assuming that high LDL-C in PBC is pri-        extended to the superior vena cava with an additional
                    marily due to the presence of LpX, which could explain      non-occluding thrombosis to the cavo-atrial junction.
                    the lack of response with Evolocumab, we detect indi-       Enoxaparin 8000 UI twice/die was re-started followed
                    rectly the presence of LpX by measuring ApoB levels.        by rivaroxaban 20 mg for 3 months. In November a CT
                    Patients with elevated Lp-X typically have low Apo B.       scan showed initial recanalization of the anonymous,
                    ApoB levels were not extremely high (160 mg/dl), not        subclavian and jugular veins, reduction of cavo-atrial
                    excluding our hypothesis. Conclusions: The only             thrombosis. Rivaroxaban is still ongoing without any
                    explanation for PCSK9-resistance in our patient is a        hemorrhagic complications. Cohort study: 59 pediatric
                    LpX-hypercholesterolemia. A limitation of our report is     patients with CAT were considered (Table 1). The most
                    that the measure of ApoB levels is not an accurate          involved neoplasms were hematological malignancies.
                    method for detecting LpX. Electrophoresis techniques,       24 events (40.6%) were cerebral arterial thrombosis;




                                                                                                          ly
                    which show directly the presence of LpX comigrating         35 (59%) were venous thrombosis (VTE, 8 cerebral
                                                                                vein, 25 systemic vein thrombosis and 2 pulmonary




                                                                                                  on
                    with LDL, are more sensitive, but they are not available
                    in all laboratories. Since studies emphasize that marked    embolism). Of 27 VTE, 15 (55%) were CVC-related,
                    hypercholesterolemia in PBC is not associated with an       19 (70%) were symptomatic. Main sign/symptoms
                    increased cardiovascular risk and the patient has not       were 4 catheter malfunction, 2 dyspnea and 13 limb




                                                                                            e
                    further cardiovascular risk factors and is refractory to    edema and pain. Antithrombotic therapy was adminis-
                    lipid-lowering therapy, we consider not necessary to
                    ahead lipoprotein apharesis in primary prevention.              us
                                                                                tered in 51/59 (86%) of patients, mainly heparin (98%)
                                                                                (low molecular weight heparin [mean dosage 162.25
                                                                                U/kg/die] or unfractionated heparin [mean dosage 19.7
                                                                                U/kg/h]). One patient was treated with warfarin (2%).
                                                                                al
                                                                                As for the outcomes, 2 patients died of CAT-related
                      PO121
                                                                        ci

                                                                                death (3.4%), 6 patients (10.1%) underwent CAT
                                                                                recurrence after a mean period of 540 weeks [IQR 428-
                                                                  er


                    DEALING WITH PEDIATRIC CANCER ASSOCIATED                    652]. Recurrence events were 3 cerebral vein and 3
                    THROMBOSIS: A MONOCENTRIC COHORT STUDY                      systemic vein thrombosis. As for bleeding complica-
                                                             m



                                                                                tions only 3 minor events were reported (5%).
                    E. Campello1, D. Tormene1, A. Francavilla2, S. Sartori3,    Conclusions: Pediatric CAT characteristics and out-
                                                 om




                    S. Sebellin3, M.F. Pelizza3, M. Nosadini3, J.N. Pin3,       come resemble those of adult CAT. Particularly, a high
                    G. Lorenzoni2, A. Biffi3, D. Gregori2, L. Spiezia1,         incidence of both arterial and venous events
                    M. Martinato2, P. Simioni1                                  was reported; hematological neoplasms and CVC
                                                                                are the major risk factors; heparin the only used ther-
                                              -c




                    1Department   of Medicine, University of Padova, Italy;     apeutic option; a high incidence rate of recurrence was
                    2Unit of Biostatistics, Epidemiology and Public Health      detected.
                                      on




                    Department of Cardiac Thoracic Vascular Sciences and
                    Public Health, University of Padova, Italy; 3Department     Table 1. Characteristics of the study population.
                                  N




                    of Women’s and Children’s Health, University Hospital
                    of Padova, Italy

                    Background and Aims: Little is known about cancer-
                    associated thrombosis (CAT) in children. We aim to
                    report a pediatric case of CAT successfully treated with
                    rivaroxaban and to describe clinical presentation, man-
                    agement and outcomes of pediatric CAT. Methods: All
                    consecutive patients admitted at the Pediatric
                    Department of the Padova University Hospital between
                    January 2007 and December 2021 with a thrombotic
                    event were retrospectively enrolled. Inclusion criteria
                    were age >28 days, confirmed acute venous or arterial
                    thrombosis, cancer among risk factors. Clinical out-
                    comes including thrombosis-related death, recurrence
                    and bleeding complications were recorded. Results:
                    Case: A 13-year-old girl undergoing systemic
                    chemotherapy via central vein catheter (CVC) for a




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 161



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    PO122                                                         ficial phlebitis was small and did not request heparin
                                                                                  therapy. Clinically relevant non major bleeding was
                                                                                  favored by concomitant anti-inflammatory theraphy.
                    VENOUS THROMBOEMBOLISM PROPHYLAXIS IN
                                                                                  Conclusions: These data suggest that critically injured
                    PEDIATRIC TRAUMA:                                             adolescent are at meaningful risk of VTE and represent
                    A MANAGEMENT ASSESSMENT                                       an important group for prophylaxis. Risk stratification
                                                                                  as poposed by the algoritm can be considered a simple
                    A.M. Pizzini1, M. Silingardi1, C. Fantoni1, C. Coniglio2,     tool to identify patients at high risk of VTE who may
                    C. Ghizzi3                                                    benefit from heparin prophylaxis. However further
                    1Department
                                                                                  studies are needed for its validation.
                                 of Internal Medicine, Maggiore Hospital
                    Carlo Alberto Pizzardi, Bologna, Italy; 2Department of
                    Anesthesiology, Intensive Care and Pre-Hospital
                                                                                  PO123
                    Emergency Services, Maggiore Hospital Carlo Alberto
                    Pizzardi, Bologna, Italy; 3Pediatric Unit, Maggiore
                    Hospital Carlo Alberto Pizzardi, Bologna, Italy               RECURRENT FAILURES IN ASSISTED
                                                                                  REPRODUCTIVE TECHNIQUES: THE FIRST
                    Background and Aims: Venous Thromboembolism                   REGISTRY
                    (VTE) is rare in healthy children (incidence around




                                                                                                          ly
                    1%), but in hospitalized patients the incidence has           E. Grandone1,2, D. Colaizzo1, A. De Laurenzo1,
                    increased over the last decades, due to improved med-         P. Totaro3, T. Bartolotti4, E. Bucherini5,




                                                                                                  on
                    ical care and radiographic detection methods. Trauma is       M. Margaglione6, on behalf of the SSC Subcommittee
                    an independent VTE risk factor in adults, with a report-      on Women’s Health Issues in Thrombosis and
                    ed incidence from less than 1% to 7.6%. In pediatric          Hemostasis




                                                                                             e
                    trauma patients the incidence is lower (0.1-1.7%) but in
                    the subset of pediatric patients with hospital-associated     1Thrombosis
                    VTE, trauma is on of the most frequent diagnoses.
                    Methods: We evalued a case series of twenty injured
                                                                                      us        and Haemostasis Unit, IRCCS “Casa
                                                                                  Sollievo della Sofferenza”, San Giovanni Rotondo
                                                                                  (Foggia); 2Ob/Gyn Department, University of Foggia,
                    children related to the Trauma Center of our hospital         Foggia; 3Santa Maria Hospital, Bari; 4Demetra Artebios,
                                                                                  al
                    over a period of about two years. The patients were           Lugo di Romagna, Ravenna; 5Angiologia e Medicina
                    stratified according to the risk assessment algorithms
                                                                          ci

                                                                                  Vascolare, AUSL Romagna, Lugo di Romagna, Ravenna;
                    proposed by JK. Petty in “Seminars in Pediatric Surgery       6Medical Genetics, University of Foggia, Foggia, Italy
                                                                    er


                    2017”. This work describes the patients characteristics
                    and the efficacy and safety outcomes related to heparin       Background: Clinical pregnancy rate among women
                                                              m



                    prophylaxis. Results: The results are summarized in the       undergoing embryo transfer (ET) after Assisted
                    Table 1.                                                      Reproductive Techniques (ART) varies from 20% to
                                                  om




                                                                                  35% per transfer and the higher is the number of
                    Table 1. Results.                                             attempts, the lower the chance of a live birth ( Harlin J,
                                                                                  et al. Hum Reprod 2002). Although the role of throm-
                                                                                  bophilia screening is very controversial in this setting
                                               -c




                                                                                  (Di Nisio M, Blood. 2011), nevertheless several
                                        on




                                                                                  Fertility Clinics test women for inherited and acquired
                                                                                  thrombophilia before ART. The benefit of LMWH in
                                                                                  improving live-births after ART has been shown irre-
                                    N




                                                                                  spective of thrombophilia status (Potdar N, et al. Hum
                                                                                  Reprod Update 2013; Grandone E, et al. PLOS One
                                                                                  2014). Because of their antithrombotic and vasodilatory
                                                                                  properties, many studies have investigated the effects of
                                                                                  low-dose aspirin or low-molecular-weight heparin
                                                                                  (LMWH) to improve ART outcomes. Two systematic
                    Prophylaxis is performed more frequently in older chil-       reviews and meta-analyses assessing the effect of low
                    dren and in puberty, confirming this group at greater         dose aspirin and LMWH in women undergoing ART
                    risk of VTE. More than two risk factors regardless of         showed that low dose aspirin was not associated with a
                    trauma, are present in children in prophylaxis, even in       higher rate of live birth, whereas LMWH could be
                    those under 13 years. The trauma severity (Injury             effective in increasing the rate of live births
                    Severity Score ISS ≥16), surgery and vascular device          (Helmerhorst FM, BMJ 2004; Dentali F. J Thromb
                    are the main risk factors for thrombosis. Anti-Xa activ-      Haemost 2011). Despite increasing knowledge on
                    ity was founded in the normal range (0.2-0.4 UI/ml) in        reproductive failures over the past decades, there are
                    nine children and in one it was 0.12 UI/ml, but the           still several aspects of clinical management, treatment
                    heparin dosage was not modified for concomitant               and prognosis with uncertainties that need to be
                    antiplatelet theraphy. The distal venous thrombosis is        addressed. We have set up the multi -centre prospective
                    likely due to incorrect risk stratificatin while the super-   registry “FIRST” (recurrent Failures in assIsted




                                                               [Hematology Reports 2021; 13:s3]                                            [page 161]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 162



         Poster




                    Reproductive Techniques ), NCT 02685800. The main             Medical University, Obstetrics & Gynaecology, Moscow,
                    objective of the registry is to investigate factors affect-   Russian Federation; 3University of Foggia; University
                    ing reproductive outcomes and assess the best clinical        of Cagliari, Italy
                    management strategies of women with recurrent other-
                    wise unexplained implantation/pregnancy failures.             This narrative review is focused on coagulation and fib-
                    Methods: We are recruiting women approaching ART              rinolysis changes in users of estrogens and progestins at
                    cycle with 2 or more implantation failures/losses of          different ages of life. Indeed, the risk of venous or arte-
                    clinical pregnancies after ART (i.e.: First, second and       rial thrombosis widely changes according to age and
                    third level techniques). Exclusion criteria are: Uterine      different risk factors throughout life. The aim should be
                    abnormalities, Hydrosalpinx, Chromosomal abnormali-           avoiding inflexible or fearful behavior encouraging cli-
                    ties in parents, Known hemorrhagic diathesis, Previous        nicians to find the best solution for the individual
                    inclusion in the study. We estimated the inclusion of         woman after a careful counseling. Search methods -
                    624 women; this estimate is calculated on the basis of        MEDLINE and EMBASE databases were searched up
                    (1) the probability of observing live births after ART        to November 2021 with the following key-words:
                    procedures (20-30%), (2) the estimate of an absolute          “estrogens” OR “progestogens” OR “combined contra-
                    increase of 5-10% in the number of live births after a        ceptives” OR “pill” OR “contraception” OR “patch”
                    prophylaxis of LMWHs (statistical power: 80%, signif-         OR “vaginal ring” OR “IUD” OR “hormonal replace-
                    icance level: 0.05). Baseline characteristics, past and       ment therapy” AND “venous thromboembolism”, OR
                    current obstetric history are obtained during routine         “deep vein thrombosis” OR “pulmonary embolism” OR




                                                                                                          ly
                    clinical follow-up or telephone interviews and recorded       “arterial thrombosis”. Articles were excluded if they: 1)
                    into a dedicated database. All women are followed until       are not closely related to the topic of the review 2) are




                                                                                                   on
                    4 weeks after the delivery or pregnancy test after ART        not published on peer reviewed journals, 3) are not in
                    procedure. Approval of local Ethics Committees from 6         English language. Estrogens and progestins can vari-
                    Centers (Fertility Clinics and Thrombosis and                 ably induce a hypercoagulable state (1,2). Furthermore,




                                                                                             e
                    Haemostasis Centers) was obtained. Results: So far, 6         they can induce a hyper-fibrinolytic state counterbal-
                    Centers have recruited 493 women. Preliminary find-
                    ings are attached. Overall, 35/86 (40.7%) live births
                    were observed after any prophylaxis (LMWH and or
                                                                                      us
                                                                                  ancing the activation of blood coagulation (3). From a
                                                                                  clinical perspective, compared to non-users, women in
                                                                                  child-bearing age using second generation pill have a
                                                                                  al
                    ASA) and 64/341 (18.8%) in absence of prophylaxis (p:         two-threefold higher risk of venous thromboembolism
                    0.000, OR: 2, 9, 95% CI 1.7 - 4.8) (Table 1).                 (VTE) while those using third / four generation oral
                                                                          ci

                    Conclusions: Preliminary data suggest that use of pro-        contraceptives have a six to sevenfold increased risk
                    phylaxis with ASA and/or LMWH can improve fetal               (4,5). In postmenopausal women venous thrombotic
                                                                    er


                    outcomes in women undergoing ART.                             risk increases with almost all combined oral contracep-
                                                                                  tives while transdermal hormonal replacement therapy
                                                              m



                    Table 1.                                                      and tibolone seem to be the safest treatments (6,7). As
                                                                                  regard to arterial thrombosis in child-bearing women
                                                  om




                                                                                  age, the relative risk decreases by lowering the estro-
                                                                                  gens dose, with no significant differences with regard to
                                                                                  progestogens in third / fourth generation pill compared
                                               -c




                                                                                  to those in second generation ones. Robust data show
                                                                                  that cyproterone acetate and drospirenone do not
                                       on




                                                                                  increase thrombotic risk in combined oral contracep-
                                                                                  tives formulations (8). Oral progestogens and lev-
                                                                                  onorgestrel Intra Uterine Device (IUD) are the safest
                                  N




                                                                                  products, since they do not increase neither the venous
                                                                                  thromboembolic nor the arterial thrombotic risk (9,10).
                                                                                  In women within 10 years of menopause onset, the use
                                                                                  of low dose of oral or transdermal hormonal therapy
                                                                                  seems to be safe with regard to the risk of cardiovascu-
                      PO124                                                       lar death and stroke (7, 11). Recently, two large clinical
                                                                                  trials (12,13) have shown that the risk of venous and
                                                                                  arterial thrombosis is similar to that of second genera-
                    DON’T BE AFRAID OF THROMBOTIC RISK OF                         tion pill, by replacing ethinylestradiol with valerate
                    HORMONES: THE IMPORTANCE OF                                   estradiol or 17 B-estradiol. This narrative review could
                    PERSONALIZED MEDICINE FOR A SAFE                              be of some help in the daily clinical practice, since it
                    PRESCRIPTION                                                  gives a comprehensive view of the balance risk/benefit
                                                                                  profile for available products. Finally, it will give some
                    E. Grandone1, F. Marongiu2, D. Barcellona3                    practical suggestions on the choice of the best hormonal
                                                                                  therapy in defined situations. Age, obesity, smoking,
                    1Fondazione   IRCCS Casa Sollievo della Sofferenza,           family history of thrombosis and known thrombophilia
                    Thrombosis and Haemostasis, San Giovanni Rotondo              are common conditions in women who plan to take hor-
                    (FG), Italy; 2The First I.M. Sechenov Moscow State            monal therapies. Although ISTH has classified the




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 163



                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    estrogen therapy as a minor transient risk factor for        thrombosis (14). In these situations, choosing the most
                    thrombosis, the synergistic effect of individual risk fac-   suitable hormonal therapy for the individual patient can
                    tors might heavily affect the risk of venous and arterial    significantly reduce the absolute thrombotic risk.




                                                                                                        ly
                                                                                                 on
                                                                                           e
                                                                                    us
                                                                                 al
                                                                         ci
                                                                   er
                                                             m
                                                 om
                                              -c
                                       on
                                  N




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SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina 164



         Poster




                                                                                         ly
                                                                                         on
                                                                                 e
                                                                            us
                                                                      al
                                                                  ci
                                                             er
                                                           m
                                              om
                                           -c
                                     on
                                N




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                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    INDEX OF AUTHORS                                          Bandinelli, B. 19, 56, 84, 155
                                                                              Banfi, C. 76
                    A                                                         Banov, L. 129
                    Abatangelo, I. 36                                         Bar-Joseph, I. 124
                    Abdelwahab, S. 27                                         Baratta, F. 150
                    Abenante, A. 157                                          Barbar, S. 88
                    Adam, E. 124                                              Barbieri, G. 16, 28, 76, 84, 154, 156
                    Adipietro, I. 116                                         Barbieri, S.S. 76
                    Ageno, W. 1, 36, 62, 63, 65, 83, 90, 91                   Barcella, L. 31, 59, 90, 117, 137
                    Agnelli Giacchello, J. 133                                Barcellini, W. 49
                    Agnelli, G. 65                                            Barcellona, D. 122, 151, 153, 162
                    Agosti, P. 51, 58, 145                                    Bardetta, M. 52, 94, 95, 117, 131
                    Agostoni, P.G. 33                                         Barinov, V. 93
                    Akula, S. 3                                               Baronciani, L. 44, 77, 120
                    Al, S. 78                                                 Barozzi, S. 111
                    Albiani, R. 28                                            Bartimoccia, S. 12, 14, 18, 153
                    Alessi, M.C. 117                                          Bartolomei, F. 9, 53, 69
                    Alfano, F. 16, 19                                         Bartolotti, T. 161
                    Alfonsi, L. 17                                            Bartuli, A. 127




                                                                                                     ly
                    Almerigogna, F. 57                                        Bassi, C. 8
                    Aloise, C. 61                                             Basso, M. 132




                                                                                              on
                    Amadio, P. 76                                             Bassotti, G. 67, 151
                    Ambaglio, C. 137                                          Battafarano, G. 45
                    Amendola, A. 134                                          Battilà, I. 69




                                                                                         e
                    Ames, P.R.J. 41, 90, 96, 153                              Batur, M. 78
                    Amitrano, M. 95, 103
                    Ammollo, C.T. 47
                    Andreini, D. 33
                                                                                 us
                                                                              Becatti, M. 6
                                                                              Becattini, C. 11, 31, 36, 40, 65, 67, 86, 151
                                                                              Becce, M. 134
                                                                             al
                    Andreozzi, G.M. 93                                        Becchetti, A. 3, 12, 21, 33, 35, 110
                    Annechini, G. 71                                          Bellesso, S. 24
                                                                         ci

                    Anselmi, F. 20                                            Benatti, S.V. 137
                    Antignani, G. 103                                         Bencivenga, C. 31
                                                                   er


                    Antonucci, E. 1, 2, 7, 24, 41, 55, 62, 63, 64, 65, 66,    Benetti, A. 70
                        83, 90, 149, 151                                      Berardi, E. 29
                                                             m



                    Apruzzi, L. 38, 138                                       Bermejo, N. 117
                    Arba, F. 16, 73                                           Bernabé-García, Á. 143
                                                 om




                    Arcaro, A. 90, 96                                         Bernardi, F. 7, 43, 45, 46, 47, 75
                    Arcudi, S. 72, 119                                        Bertelli, A. 16, 19, 56, 60, 81, 84, 139, 154, 155
                    Arcuri, P. 9, 53, 69                                      Berteotti, M. 16, 64, 82, 84, 149, 155
                                              -c




                    Argento, F.R. 6                                           Bertolani, L. 19
                    Argirò, A. 158                                            Bertoni, E. 97
                                       on




                    Ariano, A. 116                                            Bertozzi, I. 70
                    Artoni, A. 51, 58, 72, 145                                Bertù, L. 1, 2, 55, 62
                    Assanelli, S. 20, 21                                      Betti, S. 9, 53, 69
                                  N




                    Assanto, G.M. 71, 146, 147                                Bettiol, A. 6
                    Attanasio, M. 107, 109                                    Biagetti, G. 70
                    Auteri, G. 49                                             Bianchi Bonomi, A. 58
                    Autore, F. 68                                             Bianchi, M.E. 13
                    Avruscio, G. 140                                          Biasoli, C. 97, 100
                    Azarkeivan, A. 136                                        Biccirè, F.G. 150
                                                                              Bicego, E. 59, 87
                    B                                                         Biffi, A. 160
                    Baccellieri, D. 38                                        Biganzoli, M. 51, 58, 145
                    Bacci, M. 19                                              Bigossi, M. 39
                    Bacigalupo, A. 68                                         Biguzzi, E. 44, 77, 120
                    Baghaeipour, M.R. 136                                     Biguzzi, R. 97
                    Baiocchi, R. 27                                           Birocchi, S. 36, 41, 61, 89
                    Baldacci, E. 53, 102, 125, 147                            Boccafoglio, F. 35
                    Baldassarre, M.P.A. 143                                   Boccatonda, A. 74
                    Balducci, F. 36                                           Boi, L. 92
                    Balestra, D. 46, 47                                       Bologna, C. 95
                    Banchelli, F. 32                                          Bologna, G. 156




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         Indice degli autori




                    Bolognini, S. 26, 31, 59, 67, 71                         Capecchi, M. 51, 58, 72, 145
                    Bonaccio, M. 14                                          Capezzuoli, T. 28, 58, 82, 84
                    Bonaventura, A. 157                                      Capobianco, A. 13
                    Bondone, C. 28                                           Capobianco, T. 155
                    Bonomi, A. 33                                            Capone, L. 123, 128
                    Bonura, F. 125                                           Cappellacci, I. 143
                    Borchiellini, A. 28, 47, 49, 52, 55, 94, 95, 100, 102,   Cappelli, F. 158
                       117, 131                                              Cappucci, F. 110
                    Borella, E. 27                                           Caprari, R. 39
                    Borghi, M. 78                                            Caputo, D. 134
                    Borroni, E. 3                                            Caputo, R. 127
                    Borsotti, C. 20, 21, 99                                  Caramazza, D. 145
                    Boscarino, M. 78, 112                                    Cardamone, A. 98
                    Boscaro, F. 140                                          Carli, G. 24, 49, 51, 92
                    Bosch, F.T.M. 8                                          Carmina, G. 152
                    Boscolo-Anzoletti, M. 119                                Carnevale, R. 12, 14, 18, 153
                    Bosi, C. 49                                              Carotti, A. 113
                    Bosi, E. 38                                              Carpenedo, M. 49
                    Bossi, E. 89                                             Carrai, V. 49




                                                                                                    ly
                    Bottiglieri, M. 98                                       Carulli, C. 4
                    Bouslama, K. 93                                          Casazza, G. 61




                                                                                             on
                    Bowen, J. 78                                             Casini, M. 155
                    Bozzi, S. 89                                             Casolo, G. 157
                    Bozzi, V. 111                                            Casoria, A. 103




                                                                                       e
                    Bracone, F. 14, 30                                       Cassioli, G. 16, 28, 73, 82, 84
                    Brambilla, M. 3, 12, 18, 21, 33, 35, 109, 110
                    Branchini, A. 7, 43, 45, 75
                    Brioschi, M. 76
                                                                                us
                                                                             Castagna, A. 38, 138
                                                                             Castagno, E. 28
                                                                             Castaman, G. 4, 43, 44, 48, 75, 100, 107, 109, 131
                                                                             al
                    Brogni, F. 155                                           Castellani, D. 151
                    Brunelleschi, S. 18                                      Castellani, V. 12, 14
                                                                        ci

                    Bruno, B. 52, 55, 94, 95, 117, 131                       Castelvecchio, S. 136
                    Bucci, T. 41, 150, 153                                   Catalano, A. 28
                                                                  er


                    Bucciarelli, P. 1, 151                                   Cattaneo, M. 21, 89
                    Bucherini, E. 1, 41, 161                                 Cavalca, V. 114
                                                             m



                    Bulato, C. 8, 13, 34, 126                                Ceccarelli, G. 12
                    Büller, H.R. 8                                           Ceccato, D. 140
                                                 om




                    Burra, P. 126                                            Cecchetto, L. 149
                    Bury, L. 4, 49, 78, 113, 117                             Ceglie, S. 9, 53, 69
                    Buzzi, A. 101                                            Celkan, T. 78
                                              -c




                    Buzzi, S. 3                                              Cerbone, A.M. 143
                                                                             Cerchio, R. 28
                                       on




                    C                                                        Ceresetto, J. 78
                    Cacciagiù, S. 146                                        Cerletti, C. 14, 22, 30, 75, 113
                    Caiano, L. 1                                             Cerotto, V. 11
                                  N




                    Cairo, A. 51, 77, 120                                    Cesari, E. 5, 57, 78
                    Calabrese, A.M. 134                                      Cesari, F. 16, 28, 56, 60, 76, 81, 84, 118, 139, 154
                    Calabrò, P. 7                                            Cesario, V. 124
                    Calcaterra, F. 19                                        Chantarangkul, V. 23
                    Calogiuri, M. 89                                         Chatenoud, L. 27
                    Calvaruso, G. 152                                        Chavez, G.O. 147
                    Calvisi, S. 38, 138                                      Chinni, E. 110
                    Camera, M. 3, 12, 18, 21, 33, 35, 109, 110               Chiocca, S. 136
                    Cammà, G. 65, 91                                         Chistolini, A. 1, 49, 53, 71, 90, 102, 125, 146, 147
                    Cammarata, C. 104, 114, 130, 135, 138                    Chitarrelli, I. 134
                    Cammisotto, V. 12, 14, 18, 153                           Chitlur, M. 117
                    Campello, E. 8, 13, 27, 34, 85, 87, 108, 126, 160        Chiurazzi, F. 49
                    Cancellara, A. 19                                        Chiusolo, P. 68, 69
                    Cannavacciuolo, F. 103                                   Ciaccio, M. 125
                    Canobbio, I. 109, 142                                    Ciampa, A. 123, 128, 151, 153
                    Cantoni, S. 49                                           Cianciulli, M.R. 123, 128
                    Canzano, P. 3, 12, 18, 21, 33, 35, 110                   Ciardetti, N. 156
                    Capaci, V. 124                                           Ciavarella, A. 44




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                                                                   XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Ciceri, F. 38, 138                                    D
                    Ciceri, R. 19                                         D’Alessio, A. 26, 67
                    Ciminello, A. 9, 53, 69                               D’Amati, G. 12, 14
                    Cimini, L.A. 40                                       D’Ambrosio, M.F. 116
                    Cimino, E. 103                                        D’Amico, R. 32
                    Cimminiello, M. 134                                   D’Andrea, G. 90, 96, 116
                    Cini, M. 24, 93                                       D’Angelo, A. 38, 138
                    Ciotti, S. 74                                         D’Apolito, M. 116
                    Cipollone, F. 74, 156                                 D’Ardes, D. 74, 156
                    Cirillo, P. 7                                         D’armiento, M. 98
                    Ciucciarelli, L. 149                                  D’Auria, D. 143
                    Clerici, B. 61, 89                                    D’Elia, G.M. 71
                    Clerici, M. 23, 119                                   D’Errico, A. 93
                    Colaizzo, D. 66, 110, 161                             Da Lio, L. 148
                    Colombo, F. 39                                        Dagna, L. 38
                    Colombo, G. 63, 64, 90                                Dainese, C. 28, 52, 55, 94, 95, 102, 117, 131
                    Colonna, E. 57                                        Dalla Noce, C. 63
                    Colpani, P. 44, 120                                   Dalla Porta, E. 107
                    Colucci, M. 47                                        Dalla Salda, A. 140




                                                                                                 ly
                    Coluccia, A. 12                                       Dalla Valle, F. 34, 85
                    Coluccio, V. 32                                       Dalle Fratte, M. 157




                                                                                          on
                    Coluzzi, S. 134                                       Daniele, F. 100
                    Comentale, G. 98                                      De Bonis, P. 110
                    Conci, S. 35                                          De Braud, F. 26, 67




                                                                                    e
                    Concolino, P. 134                                     De Cairo, R. 159
                    Coniglio, C. 161
                    Consoli, A. 73, 156
                    Consoli, U. 49
                                                                             us
                                                                          De Candia, E. 21, 39, 115, 117, 132, 134
                                                                          De Cario, R. 16, 19, 58, 73, 76, 82, 84, 154
                                                                          De Cobelli, F. 38
                                                                       al
                    Conti, D. 37                                          De Cristofaro, R. 24, 25, 43, 45, 100
                    Conti, M. 3, 12, 21, 33, 35, 110                      De Curtis, A. 14, 22, 30, 112
                                                                   ci

                    Contino, L. 2, 55, 133                                De Fanti, A. 51
                    Cooper, D. 44                                         De Felice, F. 124
                                                              er


                    Coppens, M. 44                                        De Filippo, M. 87
                    Coppo, M. 107, 109                                    de Gaetano, G. 14, 22, 30, 75, 113
                                                           m



                    Coppola, A. 100                                       de Laat-Kremers, R. 22
                    Coppola, M.G. 95, 98                                  de Laat, B. 22
                                                om




                    Corbo, L. 137                                         De Laurenzo, A. 66, 110, 161
                    Cordisco, G. 116                                      De Leo, P. 58, 145
                    Cornacchini, S. 122, 153                              De Lorenzis, E. 72
                                             -c




                    Corso, R. 157                                         De Luca, C. 17
                    Cortellessa, F. 127                                   De Magistris, A. 25
                                      on




                    Cosi, E. 70                                           De Michele, M. 14
                    Cosmi, B. 1, 41                                       De Paolis, E. 134
                    Cosseddu, D. 54                                       De Robertis, E. 11, 31
                                 N




                    Costantini, N. 142                                    De Sena, A. 95, 98
                    Costanzo, I. 129                                      De Stefano, V. 2, 9, 49, 53, 55, 68, 69, 144
                    Costanzo, S. 14, 22, 30, 112                          De Valentin, L. 122
                    Cotelli, M.S. 63                                      Dedionigi, C. 157
                    Cottonaro, A. 3                                       Del Fattore, A. 45
                    Cozzi, G. 44, 120                                     Del Giudice, I. 71
                    Crea, E. 49                                           Del Pinto, M. 7, 15
                    Crea, R. 127                                          Del Sole, F. 150
                    Cretich, M. 12                                        del Sordo, E. 93
                    Cretu, O.C. 31, 59                                    Della Bella, S. 19
                    Crudele, F. 64, 149, 158, 159                         Della Corte, M. 105
                    Cucci, A. 20                                          Della Valle, P. 38, 138
                    Cultrera, D. 100                                      Dellanoce, C. 24, 121
                    Cuomo, V. 98                                          Dellegrottaglie, S. 98
                    Cuppini, S. 89                                        Demartis, F. 4, 48, 100, 107, 109, 131
                    Curatola, A. 115                                      Dentali, F. 39, 157
                    Curcetti, M. 116                                      Di Castelnuovo, A 14, 22, 30, 112, 156
                    Curreli, M. 124                                       Di Fiore, A. 45




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         Indice degli autori




                    Di Gennaro, L. 45                                        Fischetti, L. 110
                    Di Gioia, M. 121                                         Fjerza, R. 4, 48, 107, 109, 131
                    Di Landro, F. 9, 53, 69                                  Follenzi, A. 3, 20, 21, 46, 47, 99
                    Di Maggio, R. 152                                        Fondi, E. 39
                    Di Mario, C. 155, 156                                    Fontana, F. 91
                    Di Micco, P. 39                                          Formelli, B. 118
                    Di Minno, G. 143                                         Fortini, A. 137
                    Di Nisio, M. 83                                          Francavilla, A. 160
                    Di Pietrantonio, N. 143                                  Franceschi, L. 28
                    Di Rienzo, G. 66                                         Francesconi, P. 157
                    Di Simone, P.E. 99                                       Franchini, S. 13
                    Di Stefano, M.A. 116                                     Franchino, G. 41
                    Di Stefano, M.C. 4, 48, 107, 109                         Franco, L. 11
                    Di Tomo, P. 143                                          Franzetti, C. 88, 141
                    Di Veroli, A. 31                                         Fregoni, V. 41
                    Dirienzo, L. 47                                          Fresa, A. 68
                    Dolmetsch, R. 44                                         Fresu, L. 18
                    Dominici, M. 14                                          Frigerio, R. 12
                    Donadini, M.P. 2, 19, 36, 55, 62, 73, 91                 Fumagalli, C. 157




                                                                                                     ly
                    Donati, M.B. 14, 22, 30, 75, 113                         Fumoso, F. 111
                    Dragani, A. 114




                                                                                             on
                    Dragonetti, D. 134                                       G
                                                                             Gagliardi, M. 123, 128
                    E                                                        Gaibazzi, N. 87




                                                                                        e
                    Elmi, G. 90                                              Gaidano, G. 49
                    Emmi, G. 6, 129
                    Erba, N. 27, 63                                              us
                                                                             Galardo, G. 18, 41, 150
                                                                             Galbiati, E. 119
                                                                             Galgano, L. 109, 142
                                                                             al
                    F                                                        Galimberti, E. 59
                    Fabro, G. 157                                            Galli, E. 68
                                                                         ci

                    Facchinetti, R. 62                                       Galli, L. 38
                    Facchini, L. 51, 145                                     Galliazzo, S. 83
                                                                   er


                    Fadin, M. 34                                             Gallucci, F. 143
                    Falaise, C. 117                                          Gamba, S. 26, 31, 59, 67, 71
                                                               m



                    Falanga, A. 1, 26, 31, 59, 67, 71, 90, 137               Garcia-Leal, T. 3
                    Falcinelli, E. 4, 11, 15, 17, 31, 57, 78, 82, 113, 117   Gasbarrini, A. 93
                                                 om




                    Falco, P. 62                                             Gasparini, G. 26, 67
                    Faleschini, M. 124                                       Gatti, S. 27
                    Falini, B. 4                                             Gattillo, S. 145
                                              -c




                    Fantoni, C. 161                                          Gavasso, S. 8, 13, 34, 59, 126
                    Fantuzzi, M. 19                                          Gemmi, E. 37
                                       on




                    Farcomeni, A. 41, 150                                    Gentile, F. 90, 96
                    Fassina, A. 122                                          Gerli, S. 17
                    Fattizzo, B. 49                                          Ghali, C. 89
                                  N




                    Fazi, P. 49                                              Ghilardi, S. 76
                    Feliciani, D. 132, 134                                   Ghiotto, L. 49
                    Ferrante, N. 157                                         Ghirarduzzi, A. 140
                    Ferrara, K. 134                                          Ghizzi, C. 161
                    Ferrari, B. 51, 58, 145                                  Giaccherini, C. 26, 31, 59, 67, 71, 137
                    Ferraris, C. 28                                          Giacosa, R. 140
                    Ferrazzi, P. 73                                          Gialluisi, A. 30, 75, 112
                    Ferretti, A. 25, 53, 102, 125, 132, 134                  Giammarco, S. 68
                    Fianchi, L. 9                                            Gianfagna, F. 113
                    Fierro, T. 7, 15                                         Giangreco, A. 152
                    Filardi, N. 134                                          Giannini, M. 28, 56, 58, 82, 84
                    Filippi, L. 88, 149                                      Giaretta, A. 114
                    Fini, E. 6                                               Giarretta, I. 39, 157
                    Fino, V. 17                                              Giglio, E. 11, 31, 49, 57
                    Fiordi, T. 57                                            Giordano, G. 104, 114, 130, 135, 138
                    Fiore, M. 117                                            Giordano, P. 100
                    Fiorentini, A. 148                                       Giuffrida, G. 51
                    Fiorillo, C. 6                                           Giuliani, F. 26, 67




         [page 168]                                       [Hematology Reports 2021; 13:s3]
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                                                                          XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Giuseffi, M. 134                                              Labianca, R. 26, 67
                    Giusti, B. 16, 19, 28, 43, 56, 58, 60, 73, 76, 81, 82, 84,    Lambert, M. 117
                       118, 139, 154, 156, 159                                    Lame, D. 49
                    Giustozzi, M. 65                                              Lancellotti, S. 24, 25, 43, 45, 144
                    Gnerre, P. 39                                                 Landi, G. 95, 98
                    Goracci, L. 31                                                Landi, R. 157
                    Gorgoglione, F. 66                                            Landolfo, M. 35
                    Gori, A.M. 16, 19, 28, 56, 58, 73, 76, 81, 82, 84, 118,       Landoni, G. 138
                       139, 154, 155                                              Langer, M. 27, 63
                    Gori, F. 11                                                   Lanuti, P. 156
                    Gozzi, A. 32                                                  Lastraioli, L. 137
                    Grandone, E. 1, 66, 110, 136, 151, 161, 162                   Lattimore, S. 44
                    Graziani, M. 33                                               Laurenti, L. 68
                    Gregori, D. 160                                               Lazzareschi, I. 115
                    Gresele, P. 2, 4, 5, 7, 11, 15, 17, 29, 31, 49, 55, 57, 62,   Lazzarotto, D. 49
                       78, 82, 109, 113, 117                                      Lecchi, A. 112
                    Grimaldi, D. 131                                              Leebeek, F.W.G. 44
                    Grisillo, D. 157                                              Legnani, C. 1, 24
                    Grossi, G. 156                                                Leonardi, A. 127




                                                                                                         ly
                    Guarascio, M. 62                                              Leone, C. 115
                    Guazzini, G. 137                                              Leopizzi, M. 12, 14




                                                                                                  on
                    Guglielmini, G. 7, 11, 17, 29, 117                            Leotta, M. 103, 105, 106
                    Guida, A. 95, 98                                              Lerede, T. 41, 62, 64
                    Guida, M. 98                                                  Li, Y. 44




                                                                                             e
                    Guidetti, G.F. 109, 142                                       Li, Y.J. 93
                    Guidi, G. 146
                    Guidotti, G. 158
                    Gut, R. 44
                                                                                      us
                                                                                  Liani, R. 74, 156
                                                                                  Liberati, V. 159
                                                                                  Ligia, S. 146, 147
                                                                                  al
                                                                                  Limbucci, N. 73
                    H                                                             Limongiello, M.A. 68
                                                                          ci

                    Habib, A. 114                                                 Linari, C. 102
                    Hamuda, F. 63                                                 Linari, S. 4, 48, 75, 107, 109, 131
                                                                    er


                    Hardesty, B. 78                                               Liras, A. 3
                    Hatem, D. 114                                                 Lissandrini, L. 24
                                                              m



                    Heller, P.G. 117                                              Liu, D. 46
                    Henderson, M.W. 8                                             Locatelli, M. 38
                                                  om




                    Iacono, O. 98                                                 Lodigiani, C. 19, 73
                    Iacoviello, L. 14, 22, 30, 75, 113                            Loffreda, A. 21
                    Iannaccone, L. 90, 96                                         Loffredo, L. 12
                                               -c




                    Iannotti, W. 155                                              Loraschi, A. 100
                    Ierardi, A. 103, 105, 106                                     Lorenzoni, G. 160
                                       on




                    Ilich, A. 8                                                   Lotti Suffredini, A. 127
                    Innao, V. 49                                                  Lotti, E. 16, 19, 56, 60, 76, 83, 84, 155, 156, 157, 158,
                    Innocenti, I. 68                                                  159
                                  N




                    Insana, A. 54                                                 Lucchesi, A. 104, 114, 130, 135, 138
                    Inzitari, D. 73                                               Lucchini, E. 49
                    Iotti, M. 140                                                 Lueck, J.D. 45
                    Isola, M. 85                                                  Lugarà, M. 98
                    Iula, R. 134                                                  Luise, F. 97, 137
                    Izzi, B. 112                                                  Lunghi, B. 43, 75
                                                                                  Lupascu, A. 42
                    J                                                             Luzi, S. 122
                    Jazebi, M. 136
                                                                                  M
                    K                                                             Maccagnano, G. 66
                    Kalandadze, V. 3, 99                                          Madaric, J. 93
                    Key, N. 8                                                     Maddaluni, L. 137
                    Key, N.S. 44                                                  Madonna, P. 95, 98
                    Kura, A. 16, 19, 73, 76, 84, 118                              Maestrini, G. 53, 102, 125
                                                                                  Maggio, A. 152
                    L                                                             Maggioni, S. 73
                    La Marca, S. 112                                              Magi, A. 73, 76




                                                               [Hematology Reports 2021; 13:s3]                                               [page 169]
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         Indice degli autori




                    Magnacca, S. 14, 30, 112                                  Maugeri, N. 13, 89, 138
                    Magnoni, S. 35                                            Mavilio, D. 19
                    Maino, A. 35                                              Mazzetti, M. 129
                    Malcangi, G. 151                                          Mazzone, A. 39
                    Malfona, F. 71, 146, 147                                  McDaniel, H. 78
                    Malighetti, P. 67, 71                                     Melazzini, F. 111
                    Malvestiti, M. 5                                          Mencarini, T. 89
                    Mameli, A. 122, 153                                       Menegatti, M. 78, 136
                    Mancazzo, F. 47                                           Menichelli, D. 18, 41, 150
                    Mancini, I. 51, 58, 145                                   Merlin, S. 3
                    Mancini, M.E. 33                                          Messina, D. 101
                    Mandalà, M. 65                                            Messina, F. 29
                    Mandatori, D. 143                                         Metafuni, E. 68
                    Manfellotto, D. 39                                        Meucci, F. 158
                    Manfredi, A. 138                                          Mezzasoma, A. 49
                    Manfredi, A.A. 13                                         Mezzasoma, A.M. 17, 57, 78, 113
                    Mangiacapra, S. 95                                        Miesbach, W. 44
                    Mangiafico, S. 73                                         Mighali, P. 32
                    Mani, F. 137                                              Migliaccio, L. 41




                                                                                                    ly
                    Manieri, V.M. 146                                         Migliaccio, P. 123
                    Mannini, L. 56, 60, 158                                   Migliorini, A. 155




                                                                                             on
                    Mannucci, A. 6                                            Migliorini, S. 98
                    Mannucci, E. 157                                          Milan, M. 88
                    Mansilha, A. 93                                           Milazzo, D. 89




                                                                                        e
                    Maraglione, M. 90, 96                                     Milia, A. 81, 83, 137
                    Marchesini, E. 100
                    Marchesini, G. 51, 92
                    Marchetti, G. 89
                                                                                 us
                                                                              Minelli, M. 26, 67
                                                                              Minucci, A. 134
                                                                              Miozzo, E. 89
                                                                              al
                    Marchetti, M. 26, 31, 59, 67, 71, 89, 137, 149            Mirabella, L. 66
                    Marchini, L. 86                                           Miscia, S. 156
                                                                        ci

                    Marchionni, N. 155, 156                                   Moazezi, S. 136
                    Marchisio, M. 156                                         Mocini, A. 63
                                                                  er


                    Marcotullio, M.C. 29                                      Mohsenian, S. 136
                    Marcucci, R. 7, 14, 16, 19, 28, 37, 56, 58, 60, 64, 73,   Molinari, A.C. 100, 129
                                                            m



                       76, 81, 82, 83, 84, 107, 118, 137, 139, 149, 154,      Molineris, I. 20
                       155, 156, 157, 158, 159                                Momi, S. 4, 5, 109
                                                 om




                    Margaglione, M. 100, 116, 161                             Monahan, P.E. 44
                    Marietta, M. 32                                           Monno, A. 13
                    Marino, R. 100                                            Montanari, E. 72
                                              -c




                    Marino, S. 157                                            Montaruli, B. 54, 55
                    Marobin, M. 27                                            Montevecchi, E. 41
                                      on




                    Marongiu, F. 66, 122, 153, 162                            Morandini, R. 63, 121
                    Marottoli, V. 90, 96, 123, 128                            Morè, S. 146
                    Marrara, F. 69, 146, 148                                  Morella, P. 143
                                  N




                    Marrone, E. 143                                           Morelli, F. 49
                    Martinato, M. 160                                         Morelli, O. 67, 151
                    Martinelli, I. 1                                          Morfini, M. 75
                    Martinelli, N. 8, 75                                      Morlotti, C. 67
                    Martinelli, P. 94, 95, 131                                Mormile, M. 125
                    Martini, C. 87                                            Mormile, R. 53, 102
                    Martini, T. 97                                            Morsia, E. 69, 146, 148
                    Martino, G. 38, 138                                       Mosoni, C. 42
                    Masciocco, L. 62, 64, 149                                 Mulas, O. 49
                    Masini, F. 63                                             Mumoli, N. 39
                    Massaia, M. 55
                    Mastroiacovo, D. 1, 157                                   N
                    Mastroianni, C. 12                                        Naim, A. 129
                    Mastroianni, M. 103                                       Naim, M.C. 157
                    Mathew, P. 127                                            Nakar, C. 78
                    Mattana, M. 104, 114, 130, 135, 138                       Napolitano, M. 49, 104, 114, 125, 130, 135, 138
                    Matturro, A. 134                                          Nappi, R. 136
                    Matuška, J. 93                                            Nappini, S. 73




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                                                                         XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Natesirinilkul, R. 78                                        Patrono, C. 114
                    Negrini, A. 61                                               Patti, G. 7
                    Nencini, P. 16                                               Pavesi, F. 36
                    Neroni, S. 81, 82                                            Pavone, V. 49
                    Nicastro, M. 6                                               Pavoni, V. 37
                    Nichele, I. 24, 51, 92                                       Pazzi, M. 37
                    Nicolás, F.J. 143                                            Pecc, A. 124
                    Nicolini, E. 157                                             Pecci, A. 111
                    Nicolò, G.M. 97                                              Pedrazzi, P. 97
                    Ninivaggi, M. 22                                             Pedrini, S. 90
                    Nocella, C. 12, 14                                           Pelizza, M.F. 160
                    Nosadini, M. 160                                             Pelucchini, F. 139
                    Nosratian, V. 129                                            Pengo, M. 33
                    Notarangelo, L.D. 97                                         Pengo, V. 1, 7
                    Novembrino, C. 119                                           Peretto, L. 7, 47
                    Nuccorini, R. 134                                            Perfetto, F. 158
                    Nunziata, M. 103                                             Perin, N. 59, 87
                                                                                 Peroni, B. 61
                    O                                                            Persichillo, M. 14




                                                                                                        ly
                    Offidani, M. 146                                             Pessi, V. 19
                    Olgasi, C. 20, 21                                            Pestelli, C. 139




                                                                                                 on
                    Oliva, A. 12                                                 Peterlana, D. 35
                    Olivieri, A. 69, 146, 148                                    Petito, E. 11, 31, 49, 57
                    Olivieri, S. 102, 146                                        Petrelli, F. 26, 67




                                                                                           e
                    Oliviero, S. 20                                              Petrucci, G. 114
                    Onesimo, R. 115
                    Orlandi, S. 30
                    Orsi, R. 58, 154
                                                                                    us
                                                                                 Peyvandi, F. 23, 44, 51, 58, 72, 77, 78, 100, 112, 119,
                                                                                     120, 136, 145
                                                                                 Piacentino, F. 36, 91
                                                                                 al
                    Ostuni, A. 66                                                Piccardi, B. 16, 73
                                                                                 Picerno, A. 134
                                                                          ci

                    P                                                            Pieralli, F. 39, 137, 139
                    Paciullo, F. 15                                              Pieri, L. 4, 48, 107, 109, 131
                                                                    er


                    Padovan, L. 112                                              Pignani, S. 46
                    Pagani, P. 157                                               Pignatelli, P. 7, 12, 14, 18, 41, 64, 149, 150, 153
                                                              m



                    Pagano, L. 9                                                 Pilato, E. 98
                    Pagliari, M.T. 44, 77, 120                                   Pin, J.N. 160
                                                   om




                    Palareti, G. 1, 7, 24, 41, 62, 63, 64, 65, 66, 83, 90, 93,   Pinna, C. 21
                        149                                                      Pino, L. 127
                    Paliani, U. 11                                               Pinotti, M. 7, 43, 45, 46, 47
                                                -c




                    Palla, R. 78, 136                                            Piovella, F. 88, 141
                    Palladino, A.M. 31, 59, 137                                  Pipe, S.W. 44
                                       on




                    Palladino, G.P. 116                                          Pipino, C. 143
                    Pallotta, A. 102                                             Piromalli, A. 97
                    Palmisano, L. 98                                             Piselli, E. 17
                                  N




                    Palumbo, D. 38                                               Pitrolo, L. 152
                    Pancani, R. 41                                               Pizzini, A 83
                    Pandolfi, A. 143                                             Pizzini, A.M. 161
                    Pandolfi, M. 105                                             Pizzuti, M. 134
                    Pane, F. 49                                                  Pizzuto, A. 94, 95
                    Panepinto, L. 89                                             Podda, G. 89, 117, 145
                    Panzera, T. 14, 22, 112                                      Podda, G.M. 21, 36, 41, 61
                    Paoletti, O. 1, 63, 65, 121                                  Poggesi, A. 16, 19
                    Papa, G. 95                                                  Poggesi5, A. 118
                    Papandrea, M. 54                                             Poggi, M.M. 129
                    Parati, G.F. 33                                              Pola, R. 39, 42, 91, 93
                    Parente, R. 153                                              Poli, D. 1, 16, 27, 41, 56, 60, 62, 63, 64, 65, 81, 83,
                    Parisi, R. 41                                                    149, 151, 158
                    Pasca, S. 120                                                Pollio, B. 28, 100, 102
                    Pascale, S.P. 134                                            Poloni, A. 49
                    Pasquarella, A. 115                                          Pomero, F. 39
                    Pastori, D. 18, 41, 64, 149, 150, 153                        Pompili, M. 25
                    Patriarca, A. 49                                             Pontalto, L. 49




                                                               [Hematology Reports 2021; 13:s3]                                            [page 171]
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         Indice degli autori




                    Pontecorvo, S. 71                                     Romano, S. 12
                    Ponziani, F.R. 25                                     Romualdi, E. 19
                    Porceddu, E. 42, 91, 93                               Rosi, A. 73
                    Poretto, A. 27                                        Rossi, E. 9, 49, 53, 69
                    Porfidia, A. 39, 42, 91, 93                           Rostagno, C. 66
                    Porro, B. 114                                         Rotolo, C. 104, 114, 130, 135, 138
                    Portella, G. 27, 63                                   Rovati, G.E. 21, 35
                    Portesi, N. 121                                       Rovere-Querin, P. 13
                    Prandoni, P. 1                                        Rovere-Querini, P. 38, 138
                    Prassede, S. 145                                      Ruberto, M.F. 122, 153
                    Preti, P. 83                                          Rupoli, S. 69, 146, 148
                    Prezioso, L. 145                                      Russo, L. 2, 26, 31, 55, 59, 67, 71, 90, 137
                    Prisco, D. 6, 37, 129                                 Rustichelli, S. 142
                    Profili, F. 157
                    Proietti, G. 65                                       S
                    Proko, K. 122                                         Saccarelli, L. 11
                    Puca, I. 103                                          Sacco, M. 24, 25, 43, 45
                    Pugliese, F. 12                                       Saggiorato, G. 34, 126
                    Pulcinelli, F.M. 124                                  Sala, M. 89




                                                                                                 ly
                    Pulsoni, A. 71                                        Salvadori, E. 16, 19, 118
                    Purgatorio, G. 5                                      Salvianti, F. 107, 109




                                                                                           on
                                                                          Sammartino, A. 155
                    Q                                                     Sammicheli, L. 137
                    Quadalti, P. 40                                       Sampietro, G. 71




                                                                                    e
                    Quaglia, I. 119                                       Sanchez, M.J. 3

                    R
                    Radossi, P. 100
                                                                              us
                                                                          Sandrini, L. 76
                                                                          Santacroce, R. 116
                                                                          Santagata, D. 91
                                                                        al
                    Radu, C.M. 13, 34                                     Santi, R. 133
                    Ragazzo, S. 140                                       Santilli, F. 74, 156
                                                                    ci

                    Ramirez, G.A. 13, 138                                 Santini, P. 42
                    Ramundo, F. 9, 53, 69                                 Santonastaso, F. 30
                                                               er


                    Ranalli, P. 114                                       Santopaolo, F. 25
                    Randi, M.L. 70                                        Santoro, A. 26, 67
                                                           m



                    Raso, S. 152                                          Santoro, C. 49, 53, 98, 100, 102, 125, 132, 146, 147
                    Raza, A. 36                                           Santoro, R.C. 1, 100, 103, 105, 106
                                                 om




                    Re, R. 39                                             Sarno, L. 98
                    Recchiuti, A. 74                                      Sarti, C. 73, 158
                    Recht, M. 44                                          Sarti, L. 1, 83
                                              -c




                    Renda, D. 152                                         Sartorello, F. 34
                    Renieri, L. 73                                        Sartori, M. 87
                                       on




                    Rescigno, G. 151                                      Sartori, M.T. 127
                    Ricca, I. 28, 102                                     Sartori, S. 160
                    Riccardi, L. 25                                       Savastano, E. 105
                                  N




                    Riccioni, M.E. 132                                    Savoia, A. 124
                    Ricco, A. 49                                          Sawi, A. 63
                    Righini, M. 63                                        Scalambrino, E. 23, 119
                    Rinaldi, E. 145                                       Scarpellini, P. 38
                    Ripa, M. 38, 138                                      Scatteia, A. 98
                    Rivolti, E. 49                                        Scaturro, G.M. 76
                    Rizzi, M. 137                                         Scavone, M. 61, 89
                    Roberson, C. 78                                       Schiavetti, I. 100
                    Rocca, B. 114                                         Schiavulli, M. 105
                    Rodeghiero, F. 49                                     Schieppati, F. 26, 31, 59, 71, 90, 137
                    Rodriguez, N. 78                                      Schiera, E. 124
                    Rogolino, A. 56, 60, 81, 154, 159                     Schiralli, F. 125
                    Rogolino, A.A. 82, 84, 139                            Schirru, P. 122, 153
                    Roila, F. 65                                          Sciancalepore, P. 133
                    Roman, G. 46                                          Sciarretta, S. 12
                    Romano, C. 143                                        Scortechini, A.R. 148
                    Romano, F. 104, 114                                   Sebellin, S. 160
                    Romano, M. 74                                         Secco, C. 73




         [page 172]                                     [Hematology Reports 2021; 13:s3]
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                                                                        XXVII Congresso Nazionale SISET | Perugia | 2-5 novembre 2022




                    Sechi, L. 64                                                Tarantino, G. 69
                    Seidizadeh, O. 44, 77, 136                                  Tarantino, R. 7, 47
                    Sella, C. 52, 94, 95, 117, 131                              Tarasconi, E. 2, 55
                    Semeraro, F. 47                                             Tardugno, M. 24, 25, 43, 45
                    Senzolo, M. 66, 126                                         Tartari, C.J. 26, 31, 59, 67, 71, 137
                    Serrano, L.J. 3                                             Tedesco, S. 155
                    Serrao, A. 1, 41, 71, 90, 146, 147                          Tenori, L. 118
                    Serricchio, G. 62                                           Tesone, G. 105
                    Sevivas, T. 117                                             Testa, F.M. 43
                    Sgromo, C. 20, 21                                           Testa, M.F. 7, 45
                    Shalaby, S. 126                                             Testa, S. 1, 24, 27, 41, 62, 63, 64, 65, 121, 149, 151
                    Shapiro, A. 78                                              Thukral, N. 78
                    Shasa Quiccione, M. 75                                      Ticozzi, C. 26, 31, 59, 67, 71
                    Sibilla, S. 49                                              Tirelli, P. 95, 98
                    Siboni, S. 78                                               Tirozzi, A. 75
                    Siboni, S.M. 44, 77, 127                                    Tirrito, C. 140
                    Sica, S. 68                                                 Tiscia, G. 2, 55
                    Silingardi, M. 161                                          Tiscia, G.L. 110
                    Silvani, C. 72                                              Tocci, G. 124




                                                                                                       ly
                    Silvestri, E. 129                                           Tofanelli, L. 36
                    Silvestrini, M. 148                                         Tofani, L. 37




                                                                                                  on
                    Simeone, P. 156                                             Tolomelli, G. 145
                    Simeone, P.G. 74, 156                                       Toma, A. 64, 149
                    Simion, C. 27, 34, 85, 87, 108                              Tomasello, R. 104, 114, 130, 135, 138




                                                                                           e
                    Simioni, P. 8, 13, 27, 34, 59, 85, 87, 108, 126, 160        Tomasino, R. 125
                    Simoncini, M. 94, 95
                    Simone, P.E. Di 21
                    Siragusa, S. 49, 100, 104, 114, 130, 135, 138
                                                                                   us
                                                                                Tondi, P. 42, 93
                                                                                Tonello, D. 89, 149
                                                                                Tonetto, E. 7, 46
                                                                                al
                    Sivera, P. 55, 63, 90                                       Torlone, E. 17
                    Slawka, S. 44                                               Tormene, D. 13, 27, 34, 59, 85, 87, 160
                                                                        ci

                    Sodero, A. 16, 73                                           Torre, E. 69, 146, 148
                    Sokurenko, G.Y. 93                                          Torres, M. 78
                                                                  er


                    Sorà, F. 68                                                 Torti, M. 109, 142
                    Soriano, A. 140                                             Tortorici, E. 33
                                                             m



                    Sorrentino, S. 115, 132, 134                                Toschi, V. 151
                    Sottilotta, G. 97, 100, 101                                 Tosetto, A. 1, 24, 51, 92, 100
                                                 om




                    Spadafora, C. 103, 105, 106                                 Totaro, P. 161
                    Spanu, F. 72                                                Trabattoni, D. 110
                    Spessot, M. 38                                              Tramontano, M.L. 105
                                              -c




                    Spiezia, L. 13, 27, 34, 85, 108, 126, 160                   Tremoli, E. 18, 21, 114
                    Spinelli, D. 67, 71                                         Tresoldi, C. 38, 138
                                       on




                    Sportoletti, P. 4                                           Tresoldi, M. 38, 138
                    Squillantini, L. 58                                         Triggiani, M. 153
                    Squizzato, A. 73, 151                                       Tripaldi, R. 74, 156
                                  N




                    Stella, D. 131                                              Tripodi, A. 23, 55, 112, 119
                    Stelling-Férez, J. 143                                      Trisolini, S.M. 51, 145
                    Sterpone, P. 36                                             Trivigno, S.M.G. 109, 142
                    Sticchi, E. 28, 56, 58, 73, 76, 81, 82, 84, 139, 154, 159   Trovati, A. 83
                    Strangio, A. 103, 105, 106                                  Tufano, A. 90, 95, 98, 103, 143
                    Suraci, S. 58, 73, 76, 82, 84                               Turatti, G. 27, 140
                    Susanna, C. 35                                              Turcato, G. 88
                    Svahn, J. 129                                               Turrini, R. 122
                    Svegliati Baroni, G. 69                                     Turriziani, O. 12
                                                                                Tuttolomondo, D. 87
                    T
                    Taddei, N. 6                                                U
                    Tagliaferri, A. 100                                         Urbanek, T. 93
                    Tajana, L. 88, 141                                          Uva, A. 128
                    Tala, M. 63, 65, 121
                    Talerico, R. 39                                             V
                    Talmon, M. 18                                               Vaccari, D. 100
                    Tamburini, C. 129                                           Vaglio, A. 6




                                                             [Hematology Reports 2021; 13:s3]                                            [page 173]
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         Indice degli autori




                    Valdrè, L. 100                                          Vismara, M. 109, 142
                    Valente, P. 127                                         Visonà, A. 1
                    Valenti, R. 155, 156                                    Vitale, A. 95, 98
                    Valeri, F. 49, 52, 55, 94, 95, 102, 117, 131            Vitelli, N. 95, 98
                    Valeriani, E. 150                                       Vitrano, A. 152
                    Valesella, P. 54                                        Vitulli, A. 47
                    Valpreda, A. 55, 102, 119, 131
                    van Es, N. 8                                            W
                    Vandelli, P. 32                                         Walker, G. 20
                    Vannini, F. 60
                    Varasano, E. 4                                          Z
                    Vaudo, G. 11, 31                                        Za, T. 9, 53, 69
                    Vedovati, C. 33                                         Zaboli, A. 89
                    Vedovati, M.C. 36, 40, 86                               Zaja, F. 49
                    Venditti, M. 12
                                                                            Zalunardo, B. 83
                    Venturelli, S. 137
                                                                            Zampino, G. 115
                    Venturini, M. 91
                    Verhamme, P. 63                                         Zanatta, N. 1
                    Verso, M. 67, 151                                       Zanetto, A. 126




                                                                                                   ly
                    Vertone, D. 134                                         Zaninetti, C. 111
                    Verweij, S. 44                                          Zanofino, L.A. 37




                                                                                             on
                    Verzeroli, C. 26, 67, 71                                Zanon, E. 100, 108, 120
                    Vettorato, E. 35                                        Zarà, M. 76
                    Vianelli, N. 49                                         Zardo, L. 122




                                                                                      e
                    Vicini, R. 32                                           Zecca, C. 121
                                                                            Zerbi, A. 121
                    Vignoli, A. 71, 118
                    Villa, A. 57
                    Villa, M.R. 100
                                                                                us
                                                                            Zerbinati, P. 34
                                                                            Zocco, M.A. 25
                    Villa, S. 72                                            Zora, E. 125
                                                                          al
                    Vinci, A. 40, 67, 151                                   Zorzi, S. 24
                                                                            Zunino, F. 31
                                                                      ci

                    Viola, F. 53, 102, 125
                    Violi, F. 12, 14                                        Zuppiroli, A. 157
                                                                   er
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         [page 174]                                       [Hematology Reports 2021; 13:s3]
SISET_abstract book_180pag.qxp 27/10/22 09:09 Pagina IV




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SISET_abstrac book_cover dorso 9mm.qxp_Layout 1 27/10/22 08:35 Pagina 1




                                                                                                                                                                          Official Journal of
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                                                                                                                                                                          Società Italiana per lo Studio dell’Emostasi e della Trombosi




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                                                                                                                                           XXVII Congresso Nazionale SISET
                                                                                                                                           Perugia | 2-5 novembre 2022




                                                               Official Journal of


                                                                                 SISET
                                                               Società Italiana per lo Studio dell’Emostasi e della Trombosi
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