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Paolo Gresele
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Bleeding, Thrombosis and Vascular Biology
Platelet genetics
The amazing genetic complexity of anucleated platelets
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2022-04-01
2022-05-23
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Bleed Thromb Vascul Biol
Paolo Gresele
2022/07/15
10.4081/btvb.2022.33
Department of Medicine and Surgery, Section of Internal and Cardiovascular Medicine, University of Perugia
2785-5309
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Platelet genetics
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Margis R, Margis R, Rieder CR. Identification of blood microRNAs associated to Parkinson’s disease. J Biotechnol 2011;152:96-101.
The amazing genetic complexity of anucleated platelets
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                                                                                                               INVITED REVIEW

The amazing genetic complexity of anucleated platelets
Loredana Bury, Paolo Gresele

Department of Medicine and Surgery, Section of Internal and Cardiovascular Medicine, University of Perugia, Italy




    Although anucleated, platelets have a rich and com-                      growing in the last 10 years, particularly after the advent
plex transcriptome which includes, besides coding mes-                       of next-generation RNA-sequencing (RNAseq) tech-
senger RNAs (mRNAs) and pre-mRNAs, also non-coding                           niques. RNAseq provided new insights into the platelet
RNAs (ncRNAs) such as microRNAs (miRNAs), circular                           transcriptome, showing that about 8,000 transcripts are
RNAs (circRNAs), long noncoding RNAs (lncRNAs) and                           present in platelets,1,2 and allowed to identify mutations
YRNAs. Actually, platelets are probably the primary                          in NBEAL2 as the genetic cause of the Grey Platelet Syn-
source of ncRNAs in the circulation. These wide array of                     drome, an inherited platelet disorder that had remained
RNAs derives from the platelet precursors, megakary-                         without a causative gene for more than 30 years.3




                                                                                               ly
ocytes, but also exogenous RNAs present in the circula-                          Several studies have shown protein synthesis by anu-




                                                                                       on
tion are taken up by platelets. This diverse and dynamic                     cleated platelets, both basally and in response to activa-
transcriptome enables platelets to synthesize new proteins                   tion,4-6 and it was shown that neo-synthesis of some
which modulate their functions, both constitutively and                      selected proteins is regulated by signal-dependent pre-




                                                                                 e
upon activation, but also to modify the phenotype of other                   mRNA splicing.7,8
cells with which platelets interact.
    The field of platelet transcriptomics has been rapidly
                                                                            us   Platelets also contain ribosomes, that are bound to
                                                                             translating mRNAs protecting them from degradation,
                                                                             and thrombin increases ribosomal occupancy on mRNAs,
                                                                            al
                                                                             according with its ability to trigger protein synthesis in
                                                         ci

                                                                             platelets.4,9
  Correspondence: Paolo Gresele, Department of Medicine and Sur-                 Moreover, it was shown that mRNAs are differentially
                                                       er


  gery, Section of Internal and Cardiovascular Medicine, University
                                                                             sorted from megakaryocytes to platelets rather than ran-
  of Perugia, Centro Didattico Edificio B piano 1, 06132 Perugia,
                                                      m



  Italy.                                                                     domly distributed to them,10 suggesting that platelet pro-
  Tel.: +39.0755783989 - Fax: +390755716083.                                 tein synthetic capability and thus the platelet function
                                          om




  E-mail: paolo.gresele@unipg.it                                             phenotype, are regulated by the environment in which
                                                                             megakaryocytes mature.
  Key words: Platelet transcriptome; RNA; Platelet genetics.
                                                                                 miRNAs, short noncoding RNAs which regulate gene
                                    -c




  Contributions: LB and PG wrote the manuscript; PG critically re-           expression post-transcriptionally, represent another mech-
                           on




  vised the manuscript. LB made the literature search.                       anism of protein synthesis regulation in platelets. The
  Conflicts of interest: The authors declare no potential conflict of
                                                                             platelet miRNome is modified upon in vitro activation and
                                                                             in vivo in disease conditions, and changes of the platelet
                     N




  interest.
                                                                             miRNome affect in turn the platelet proteome.11,12 The role
  Funding: None.                                                             of circRNAs, lncRNA and YRNAs in platelets is still
  Received for publication: 1 April 2022.
                                                                             under investigation. circRNAs enrichment in platelets can
  Accepted for publication: 9 June 2022.                                     be interpreted as a signature of transcriptome
                                                                             degradation,13 but some circRNAs are selectively released
  Publisher’s note: All claims expressed in this article are solely those    in extracellular vesicles and their role is still unclear.14 Fi-
  of the authors and do not necessarily represent those of their affili-
                                                                             nally, lncRNAs and YRNA are abundant in platelets and
  ated organizations, or those of the publisher, the editors and the re-
  viewers. Any product that may be evaluated in this article or claim        might be involved in platelet reactivity.15,16
  that may be made by its manufacturer is not guaranteed or endorsed             Studies on platelet transcriptome have led to signifi-
  by the publisher.                                                          cant advances in the understanding of the role of platelets
  ©
                                                                             in health and disease. Indeed, in response to inflammatory
   Copyright: the Author(s), 2022
  Licensee PAGEPress, Italy                                                  signals, invading pathogens, cancer, or other stressors, the
  Bleeding, Thrombosis and Vascular Biology 2022; 1:33                       platelet transcriptome changes dynamically. For instance,
  doi:10.4081/btvb.2022.33                                                   the platelet transcriptome is altered in inflammatory dis-
                                                                             orders associated with increased cardiovascular risk, such
  This work is licensed under a Creative Commons Attribution-
  NonCommercial 4.0 International License (CC BY-NC 4.0).                    as sickle cell disease and systemic lupus erythemato-
                                                                             sus.17,18 The expression of several transcripts involved in

                                              Bleeding, Thrombosis and Vascular Biology 2022; 1:33
32                                                  L. Bury and P. Gresele




oxidation, immunoregulation and stress response was re-           fections, for example HIV, that is associated with an in-
ported to be higher in platelets from ST segment elevation        creased cardiovascular risk and hyperreactive platelets.26
myocardial infarction (STEMI) compared to non-ST seg-             On the other hand, anti-viral immune genes are upregulated
ment elevation myocardial infarction (NSTEMI) pa-                 in platelets and megakaryocytes in response to dengue and
tients,19 providing new insight into the pathogenic role of       influenza infection, possibly limiting viral infection.27
platelets in acute coronary syndromes.                            Platelet involvement in viral infections has led investigators
    Aging, a condition characterized by an increased car-         to search for viruses in platelets. The entry of SARS-CoV-
diovascular risk and chronic inflammation, is associated          2 in platelets is still debated: some groups described the
with alterations of the platelet transcriptome. Using sin-        presence of the viral RNA in platelets,28,29 while others did
gle-cell RNA-seq of megakaryocytes, Davizon-Castillo              not detect it,30 or detected it in very few samples.24 Other
and colleagues showed a reprogramming of the inflam-              viruses, namely HIV,31 influenza32 and Dengue,33 enter
matory, metabolic, and mitochondrial gene pathways in             platelets with the latter also stealing the platelet transla-
aged mice, detected also in circulating platelets, and as-        tional machinery to replicate and produce new virions.33
sociated with increased TNF-α.20 Another recent study el-             The platelet transcriptome is also altered in patients
egantly showed numerous differentially expressed                  with cancer, and this has been exploited to show that the
transcripts in older healthy adults in respect to younger         platelet transcriptome can serve as a diagnostic and prog-




                                                                                            ly
subjects and observed that variations of the platelet tran-       nostic platform. The uptake by platelets of tumor-associ-




                                                                                   on
scriptome influence platelet-leukocyte interactions and           ated biomolecules, mainly circulating tumor-derived
downstream cytokine synthesis contributing to chronic in-         mRNA, has led to the concept of “tumor educated
flammation.21                                                     platelets”, i.e. platelets with a highly dynamic mRNA




                                                                              e
    Moreover, RNAseq of human and murine platelets                repertoire with potential importance for cancer diagnos-
showed that the platelet transcriptional and translational
landscapes are significantly altered in sepsis, and that up-
                                                                        us
                                                                  tics. Indeed, the analysis of the platelet transcriptome can
                                                                  discriminate patients with cancer from healthy controls,
regulation of ITGA2B is associated with higher mortality22        predict the localization of the primary tumor and provide
                                                                 al
providing novel evidence that upregulation of protein syn-        information on the molecular tumor subtypes, thus guid-
                                                     ci

thetic events in platelets may play a pathogenic role in          ing clinical diagnostics and therapy selection providing
sepsis. A recent study23 confirmed the differential expres-       from a simple peripheral blood sample what has been de-
                                                   er


sion of several genes in platelets from sepsis patients com-      fined as a “liquid biopsy” of the tumor.34 Similarly, it was
                                                  m



pared to controls and showed that upregulated genes were          recently shown that the platelet transcriptome may help
mostly related to catabolic processes and protein transla-        to predict the fibrotic progression in myeloproliferative
                                         om




tion. The Authors also showed that patients with sepsis           neoplasms.35
have increased levels of immature platelets and a less de-            Post transcriptional events in platelets also regulate
graded platelet transcriptome, as indicated by decreased          key proteins related to neurodegenerative disorders. For
                                    -c




levels of circRNAs. The biological implications of the in-        instance, the amyloid precursor protein (APP) is present
                             on




creased platelet turnover remain unclear, however this            in platelets and is encoded by platelet mRNA.36 APP is
study first suggested that increased protein translation is       the metabolic precursor of amyloid Aβ peptides whose ac-
a characteristic mechanism of systemic inflammation.23            cumulation in brain parenchyma and cerebral vessel walls
                        N




    Intriguingly, there is a large overlap of the platelet        is correlated with the onset of Alzheimer disease but also
transcriptome changes observed either in sepsis or in             plays a role in platelet function.37 Recently differential
COVID-19.22-24 Distinctive changes in the gene-expres-            spliced platelet mRNA was reported to enable blood-
sion profile of circulating platelets of COVID-19 patients        based diagnosis of multiple sclerosis.38 Finally, elevated
have been shown involving the pathways associated with            expression of the platelet-related miRNAs miR-22-3p,
protein ubiquitination, antigen presentation, and mito-           miR-146a and miR-155 were found to be a possible
chondrial dysfunction,24 suggesting a drastic change in the       Parkinson’s Disease-specific miRNA signature.39
redox balance and in platelet metabolism leading to                   New discoveries on the genetic complexity of human
platelet activation and to the high thrombotic risk of            platelets are continuously made. A recent innovative re-
COVID-19 patients. Another study similarly found in               port showed that platelets possess endogenous retro-tran-
platelets from COVID-19 patients differentially expressed         scriptase (eRT) activity that regulates protein synthesis by
lncRNAs involved in the regulation of hemostasis,                 generating RNA–DNA hybrids which block translation.
platelet activation and the immune response,25 further con-       Inhibition of platelet eRT activity increased integrin acti-
firming that platelet hyperactivation and immune-throm-           vation in vitro and thrombosis in vivo suggesting that in-
bosis are major players in COVID-19 complications.                hibition of platelet eRT activity may contribute to the
    Differential expression of platelet genes involved in         increased risk of thrombosis observed in people with HIV
leukocyte activation has also been shown in other viral in-       treated with RT inhibitors.36

                                     Bleeding, Thrombosis and Vascular Biology 2022; 1:33
                                         The amazing genetic complexity of anucleated platelets                                      33



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landscape of platelets will probably reveal unexpected                     tional profile and protein expression in patients with sys-
new perspectives.                                                          temic lupus erythematosus: upregulation of the type I
                                                                           interferon system is strongly associated with vascular dis-
                                                                           ease. Blood 2010;116:1951-7.
                                                                       19. Eicher JD, Wakabayashi Y, Vitseva O, et al. Characterization
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