All right, is this on? Yeah, it seems like we're OK. Hey, John, thanks for having me here and thanks for the kind introduction. Really excited actually to tell this group about some of our work that's sort of connecting ENCODE-like assays to really clinically entrenched problems, and in particular in the area of cancer stem cells and glioblastoma.  And despite resection of the tumor and radiation and chemotherapy, the median survival is very short and is almost uniformly fatal. So there's this lot of evidence that not all cells in glioblastoma are created equal, but rather there is this subpopulation of very aggressive stem-like cells in glioblastoma that propagate the tumor. So by the time you've resected this lesion here, there's actually stem cells, tumor stem cells, that have sort of infiltrated throughout the brain, right? And these seem to be resistant refractory to therapy. This is sort of a model of cancer stem cells. I don't mean to say that all cancers have stem cells, and it's a really controversial area, but in glioblastoma, there is evidence that not all cells are alike and that there are these purple cells that can both self-renew and propagate tumors. And for the purpose of this talk, I'll call them cancer stem cells or gliomastem cells, one can model these cells, and this is what we do. We actually take human tumors resected at MGH in surgery, and we can either expand them in stem-like conditions, in serum-free conditions, as gliomospheres. So these grow up spheres. They look a lot like neural stem cells that you might be isolating from stem cells or something. But these gliomospheres, they express CD133, which is a canonical marker of the stem-like state, and they're very aggressive. So you can put as few as 50 of these cells into a mouse brain and orthotopic xenotransplantation, and they'll cause a tumor. So these are tumor-initiating and they have tumor-propagating cells. You can take the same human tumor and you can expand cells in serum. And these grow a cell line, kind of the conventional type of cell line that we all might work with. These cells, though, they don't express CD133. And you can put hundreds of thousands or millions of these cells into a mouse, and they won't do anything. The mouse is just fine. These cells won't propagate tumors, and they lack stem-like characteristics. You can differentiate these cells. So you can put these cells into here, into the serum conditions, they'll differentiate, and they're no longer tumor-ogenic. So there's this directional thing that these are sort of a primitive state, and they will differentiate. But you can't go back. You take these cells in serum, these non-stem cells, they won't go back to this initial state. They basically won't proliferate in the gliomospheres. So this was very interesting to us, and led us to this question of whether we could reprogram these differentiated cells into tumor-propagating gliomospheres. So we basically set up this strategy where we were going to look at the enhancer landscapes and the transcription factors, and then we would sort of test the ability of these TFs to reprogram these cells into the gliomospheres. So whether we could identify something like a TF code for glioblastoma. So this is just very familiar now to this group, but this is basically, I think, a pretty striking picture that says, well, if you take three different patients' tumors and you expand them in the stem-like conditions, you can see that the SOX2 locus is very active, and there are these sort of H3K27 acetylated peaks in the chip-seq profiles that are enhancers in the SOX2 locus. But if you take these same tumors and you expand them in the differentiated cell line conditions, this is a very quiet locus.  Conversely, if you look at some locus like BMP4, which is associated with differentiation, you see precisely the opposite. So in these cases, it's not which patients' tumor you're looking at. It's whether you're isolating and expanding a stem-like population or a differentiated population that is underlying at least this landscape. But I think you can see it's more striking when you now, each little row here is a different candidate enhancer or K27 acetyl peak in the genome. You can see there's a large group of shared enhancer-like elements shared between the stem-like cells and the non-stem cells. These have motifs for some cell cycle regulators, right? These guys are all proliferating. But what we're interested in here are these stem-like cell specific peaks. You can see these guys are all active in the stem cells and not in the differentiated cells. Again, motif analysis identifies HLH motifs and SOX motifs suggestive neurodevelopmental program. So then what we can do is we can look at RNA-seq data and we basically identified a set of about 20 different transcription factors that are predicted to bind to these motifs and that we posited might be explaining this different signature in the CSEs, in which are expressed specifically in the CSEs. So we went back and took the differentiated cells and we over-expressed each of these transcription factors one by one to ask whether they could reprogram, right? Kind of like the Yamanaka experiment. But we're looking for cancer stem cells. So basically if you take the differentiated cells and you try to expand them in sort of the spherogenic conditions, basically nothing happens and you can see that here. You get zero sphere forming cells. But if you now start adding these different transcription factors, there are a couple that kind of lit up and you could sort of use your, if you were very optimistic, you might guess that POW3F2 gives you a little bit of something growing in the plate. No real CD133, but we thought we might be moving in the right direction. So we started trying combinations of factors. Sox2, you get a little more sphere formation and so on. Ultimately by sort of iterating between additions of factors and then looking at the enhancer maps that we were getting, we were able to sort of come up with this cocktail and ultimately define four TS, POW3F2, Sox2, Sal2, and Olig2, that when you added them to the cell lines, they would reprogram into spherogenic stem-like cells that could propagate tumors.  Okay, so we'll call these induced GSCs. As you might hope, you can see that these CSC specific enhancer elements, after you've reprogrammed the cells with the core TS, they get K27 acetylation. So you're really reprogramming also the chromatin landscapes. The chromatin landscape in this case is a pretty good surrogate for the functional state of the cells. Another, I think, pretty critical data point here is that you only require transient expression of the core TS. So in some reprogramming or directed differentiation experiments, the factors are required continuously to maintain a new state. So that raises the question of whether you've really reprogrammed, whether you've really achieved an epigenetic self-sustaining transition.  We can tell this by looking at the three-prime UTRs. And the native loci for these four core TS, actually come on, you can tell this by looking at the RNA expression patterns. And you can also tell this by looking at the loci and you can see that the enhancers in the PAL-3F2 locus, the enhancers in the SOX2 locus, right? And all of them are coming on, right? So you sort of reprogram these guys and they're now self-sustaining. And part of how you know this now too is that you can now switch these IGSEs back into serum and they'll readily differentiate and they'll totally shut off the core TFs. So it's telling you you have an endogenous regulatory program now that you've got going. So how might this be useful to understand glibostoma? Well, for one thing, we could now go back and map the direct targets of these core TFs, which we, well, PAL-3F2, SOX2, SOX2, OLG2 by chip-seq. There's a great deal of overlap. They're sort of binding to each other's loci and we think sustaining their expression. I can tell you that all four of these guys are essential for tumor propagation of the GSEs. But more than that, when you now infer the downstream targets of these core TFs, here are the core TFs and you can use the binding patterns and expression patterns to infer their downstream targets. I can tell you that essentially everything we've looked at and then tried to knock down in the GSEs has turned out to be an essential factor. So this is sort of like, you know, there's various proposals out there, whether you use super-enhancers or specific TF signatures or what, here's an example where by knowing the core TFs that are driving the circuitry and examining their direct downstream targets, you now have a set of regulators that seem also to be essential for the tumor-propagating state. We were particularly interested in ASCAL-1, which is a transcription factor that it's a neurodevelopmental TF that's activating wind signaling in the GBMs. As well as the R-Core II histone dimethylase complex, which actually three or four different members of this complex are direct targets of the core circuitry. So in a sense, knowing this core circuitry, now, like knowing the core circuitry of IPS cells, told you a lot about the pluripotent regulatory circuitry, I think we've learned a lot about the GSE circuitry by using sort of familiar ENCODE type approaches and combining them with reprogramming approaches to define the circuitry. So, but I kind of told you this was at some level, it's a very fundamental project, I think, but it's clinically driven. We're doing this because we want to understand the nature of these cells that are propagating tumors in humans. So the question then is, what's the status of these core TFs in primary glioblastomas? So here we wanted to, we actually did a series that we did flow cytometry as well as immunofluorescence analyses. We labeled antibodies that recognize each of the core factors. We also used CD133 antibody. And we stained slides of primary glioblastomas from patients. And we can see there's a great deal of heterogeneity, as you might expect, glioblastomas are notoriously heterogeneous tumor, variable expression of the core TFs, but there clearly are a subpopulation of cells, the pink ones here, that we know are co-ordinately expressing all four of the core TFs in the tumors. And when we do flow cytometry, we look at all the markers. These guys, I can tell you are also CD133 positive, which is one of the markers that enriches for tumor propagation. So it's very likely that there are surrogates within primary tumors that look very much like the stem cells that I just sort of functionally described in our model. And such as shown here. We've also looked at single cell data in another way, and that is by single cell RNA-seq. So by single cell RNA-seq, again, of primary tumors, one can learn a little bit about the regulatory programs in the primary tumors.  Except I wanna tell you one interesting property. We looked at a few hundred cells from a few different glioblastomas. We saw a lot of heterogeneity within each tumors. There's a variety of transcriptional programs. These tumors at the RNA level, as you might expect from that last picture, are very heterogeneous. But what, we wanted to take this opportunity to ask about the stem-like transcriptional programs and its presence in primary GBMs. So we defined a stem-like signature based on, which is composed of genes that are expressed at low levels in these differentiated cell lines, okay? But are highly expressed in the GSC, the stem-like model that I talked about earlier in the talk. And then we asked, what is the expression pattern of these genes in single cells from a person's tumor? So this is just one tumor from a patient, and we looked at about 70 different cells. And I think you can see that there is a population of cells at the right side of the slide here that are highly expressing a lot of the genes in the signature, okay? So together with the last picture, where I said, well, we've got the core TFs, and we've got a protein level analysis. So we've got stem-like cells that emulate our model in tumors. Now I'm telling you that we've also identified cells with a transcriptional signature. It looks a lot like what our model looks like, okay? I'm telling you all this, but now I'm gonna tell you about a little caveat, okay? The problem was that if you take these cells, and you now ask about their cell cycle signature, turns out they're not cycling. They're totally dormant and quiescent, okay? So we have this model of CSEs with the right neurodevelopmental programs that seems to be emulating something in a tumor, yet our CSEs are proliferating like mad, like probably most of the cultures that people work on in this room, because if a culture's not proliferating, it's hard to work with. But the real in vivo GSEs with this developmental program seem to be out of cycle and pretty much quiescent and dormant, okay? So this presents a bit of a challenge to us as we think about modeling and targeting, okay? I'm gonna tell you now how we're trying to get at this population. I'm gonna come back to the population in a second, but I need to just sort of talk for a minute about therapeutics, okay, before I get back there. Okay, so receptor tyrosine kinases. So RTK inhibitors, of course, have been a dramatic advance in cancer research and Brian Drucker and Charles Sawyer's and others have shown that this miraculous inhibitor of BCR-Able practically cures CML. This RTK inhibitor basically ablates this whole population and this whole tumor. So this led to a lot of excitement and now RTKs are being used in many settings. GBMs, gliomas seem like a perfect setting to use RTK inhibitors, right? Look at the genetics of gliblastoma. You've got EGFR amplification, a huge proportion, you've got PDGFR amplification and others.  So of course this led a lot of excitement to then go in the clinic and try RTK inhibitors that could target these genetic lesions and hopefully have an impact on gliblastoma. They haven't done anything in the clinic, though, right? They haven't changed the course of this disease whatsoever. So we wanted to understand this better and you'll understand where I'm going in just a minute. But we took one of our stem cell models, one of these proliferating GSC models and we took in particular one with a PDGFR amplification. And we treated it with the satinib, which is a really good PDGFR inhibitor. You can see that the satinib, this particular population is very sensitive to satinib, pretty low concentrations of this inhibitor kills the cells. This is in contrast to a couple other glioma lines. So if you take a glioma stem cells from an EGFR amplified one or a MIC amplified line, they don't care about PDGFR, so they're insensitive to satinib, okay? But look at this. Even in these cells that are supposedly totally dependent on PDGFR, there is this sort of window here where about five to 10% of the cells just won't die and they remain viable. If you culture them out for a while, they actually start proliferating very slowly, okay? And they're quite insensitive. We call them persisters. I'll call them GSC8 persisters or whatever persister stem cells. They proliferate slowly, but they're pretty much impervious to very high concentrations of this satinib. So this cell within this genetically amplified PDGFR dependent population, these cells can acquire a different state that is independent of the RTK. So this is just showing you that we take the naive cells and we can culture them in the presence of this satinib and over time we kind of acquire this slow-cycling population, okay? This is a little bit analogous to persisters that have been defined in lung, melanoma, breasts in other settings. One interesting thing is it seems to be a reversible because we can remove the pressure of the drug and these cells will go back to their initial state where they're proliferating very rapidly and PDGFR dependent, okay? It's kind of a reversible persister state. Okay, this I think was pretty interesting so we can look at the regulatory elements of the different cell states, right? So this is all from the same GSEs. Here are the naive sort of proliferative GSEs and here are the persisters, right? That after they're going, they're cycling very slowly, they're PDGFR independent. You can see enhancers that are common in various different patterns and then there's this big group here that seems to be quite a bit stronger in the persisters. I think what's kind of striking about this is if you go back and look at the tumors, the tumors actually look much more like these persisters. So in a sense, these RTK-inhibited persister cells that are very slow cycling actually look a lot more like the primary tumors of the GSE, the primary glioma tumors, okay? So that we might be doing a better job of emulating them. We want to go back and understand a little bit their circuitry and their regulation. On many levels, these persister cells look much more like tumor cells and they look in particular like those tumor cells that are quiescent or non-cycling. And I'll just give you one example that we were kind of excited about that's I think relevant to this conference. This is kind of a complicated graph, but basically we're comparing the primary tumors, different cells in the primary tumors and at the top here are genes that are associated with the quiescent tumors. They're anti-correlated with cell cycle. Here we're sort of comparing the naive GSEs to the dormant GSEs, and here are genes that are high in the quiescent drug tolerant ones that we selected. So things up here are high in the dormant non-cycling tumor cells and they're also high in the drug tolerant non-cycling GSE model. What are the genes? Well, we were pretty struck that this is just packed full of KDMs, lysine dimethylases, genes that encode these enzymes that remove the methyl marks off of histones. You might notice that in contrast, the methyl transferases actually are down-regulated in these drug tolerant cells. We thought this was pretty interesting here that these KDMs are being up-regulated. In fact, KDMs have been, particularly at KDM5 here, has been previously implicated in drug tolerant cancer cells. Let's see. I'll tell you just, I don't have a slide here, but we spent a while looking at KDM5 actually and it turns out that KDM5 does not seem to be the critical player here. We can inhibit KDM5, we can knock it down, we can actually pharmacologically inhibit it, but the cells don't care. The persister cells don't care. So they don't seem to be dependent on KDM5. This is the one that removes lysine form ethylation. Okay. But there's another set and there are these KDM6 and KDM3 families that remove repressive marks that actually attract, seem to be a little more interesting. I'll tell you why. First of all, if you look at K27ME3, repressive sort of polycomb peaks in the naive cells, you see pretty good signal. And then as these cells transition into this drug tolerant persister state, there's a global erasure of K27ME3. You can actually see that, let's see, if I'm gonna highlight it, you can see that here in the SOX2 locus where you see pretty good signal for K27ME3 in the naive and then it kind of goes away a little bit in the persisters, but it's a very global effect. So you're erasing this repressive mark. You're doing the same thing, maybe equally dramatically to K9ME3, another repressive mark, and I think you can see a region here where you've got some pretty strong K9ME3 and the persisters sort of have erased that and it's very flat. There's a KDM6, which is the one that ruins K27ME3, there's an inhibitor, GSKJ4, it's not a perfect inhibitor, but it actually seems to work pretty well because if you look at the naive cells compared to the persister cells and then the persister cells treated with the drug, the K27ME3 comes back up. So the drug sort of blocks this demethylation effect. And actually what was really striking is that these persisters are really in red here. You can see that these long-term persisters are very sensitive to GSKJ4. So they're very sensitive to an inhibition of the KDM6 at very low concentrations, suggesting that this erasure of these repressive marks may be essential for these cells to transition into this drug-tolerant quiescent state. So why do you need to do this? What's going on? Well, the persister cells have an altered developmental program, as you might suspect. So this is just looking at comparing naive to persister for K4 methylation and for K27 acetylation and for K27ME3. And we see a number of loci where there is K27ME3 sort of silencing genes in the naive cells. And then this gets erased in the persisters and up comes K27 acetyl, up comes K4ME3. We looked in general at the pathways. There are a couple of things that come on. One is very primitive neurodevelopmental genes that are associated with primitive neural stem cells. The second is notch target genes. And in fact, there's some very interesting literature in the neural development field that suggests that in radial glial development and very primitive embryonic development of neural stem cells in their transition to transient amplifying cells, notch and RTKs are sort of antagonistic in having a complex interaction. So the most primitive neural stem cells that are quiescent have notch. The transient amplifying cells get the signal from EGFR that causes them to rev up and proliferate a little bit. This is happening in a very narrow window of development but we think this may be emulated in glialostoma. And in fact, if you now go back and think about notch signaling, you can see that notch signaling is actually activated in the persisters. You can just do Western blots to stain for active form of notch. You can see old gene expression signature associated with notch signaling. And the persister cells but not the naive cells are very sensitive to gamma secretase inhibitors which target notch signaling. So this kind of leaves me here as I'll wrap up with this picture here. We think that the persister cells are sort of emulating a quiescent neural stem cell state whereby we're actually starting with these naive GSEs that are proliferating an RTK dependent but through demethylation and perhaps a resetting of their epigenetic state, we get these persister cells that are very slow cycling. They upregulate very primitive stemness genes and they're notch dependent. As I mentioned, this parallels an event or sort of inversely parallels an event in normal development whereby neural stem cells that are quiescent have notch signaling and switch to an amplified state with RTK signaling. We're very interested in how KDMs are working, we think, to sort of help allow the transition between states or back to this primitive state and allow particularly a ratio of repressive marks or repressive state can allow the right enhancers to come on and sustain the state. We're also very interested in whether now, given the observation that the primary tumor cells look a lot more like this persister state and that they're very slow cycling, suggest that perhaps notch inhibitors or better yet inhibitors of some of the chromatin enzymes might synergize with RTKs and give you a better chance of targeting some of these glioma cells. So my final slide, I've shown you that enhancer landscapes distinguish these tumor propagating GSEs from conventional cell lines that don't propagate tumors and don't have this functionality. I've shown you that we can identify core TFs that are sufficient to reprogram GSEs and these sort of define, their targets define essential drivers of the stem-like state. I've also told you about that there's a large-scale erasure of repressive marks seems to allow GSEs to adopt a more quiescent drug-tolerant state that we think is emulating the primary tumors and how this may have implications for glioma therapy that targets this other population, which we think is completely missed by existing therapies that all of them, because they all target proliferative cell states. Knowledge, Maria Suva is now a faculty at MGH formally, a postdoc in the lab, led the reprogramming work, Brian Lau, Jim Severs have been working on some of this, the persister cell story. The single cell work was a collaboration with the Thai Taroosh and Aviv Reghev at the Broad Institute in the Claremann Observatory and with MGH neurosurgery and Chekepstein and Noam Shores at the Broad have been critical for a lot of the chromatin mapping work that we do. Glad to take questions. Dependent pathways for these GBM cells, are you saying that now there is an epigenetic understanding or basis for oncogene addiction that you're able to target with? Some of these drugs? Let's see, so I think there's a point to this, that when cells become dependent on oncogene for a particular state, they will respond favorably. And I think that model holds very well for that 8% of cells that are proliferating rapidly in the tumor. The model doesn't hold so well for the other cells in this tumor that are out of the proliferative state. CML, CML oncogene, it works perfectly.  There's about 0.1% of the cells still in the bone marrow. You probably need to give them Gleevec for a while until those sort of peter out, right? But it's a cure almost. Gleevec is almost the opposite, right? That fraction of oncogene-addicted cells is that 8% that's proliferating, where there's 92% sitting in this dormant reservoir or something like that. So the model's great, it just depends on the tumor, and a lot of tumors have a very high fraction of dormant cells in this other state, other epigenetic state that can transition, and that poses a huge problem to using RTK inhibitors on their own in the clinic. You have to combine it. Great talk, quick question in regards to the state of your persistent cells. Do you think they're under epigenetic instability? So the global reduction in K27ME3, for instance, do you think that that's true of all cells? Or if you do IHC, you see that there is still a variability in some having high levels, some having low levels, and by the chip, it all looks down because the signal from all over is negated. Could it be that you have a massive amount of heterogeneity? I don't think that, I mean, it's certainly possible, technically, and in the tumors, I can't even say what they look like in the tumors. But in the actual persistent population that we're studying, where I showed those things being erased, I think if there was a lot of epigenetic variability from cell to cell, there'd be some regions with middle levels and things like that, but in regions that are really repressed in deserts where K27ME3 is sitting, it shouldn't be coming on and off and it's gone. So the fact that those deserts also get totally erased and there's no rationale that you think some population of cells is expressing in the desert, tells me it's a global effect hitting all the cells in our model. Again, I don't know what's going on in the tumor, it's very hard to work with single cells from the tumor in terms of understanding their chromatin, but it's a good question, I mean, we need to try to understand something. Great, thanks.


Yeah, so we're going to talk about pizza farming. We don't have a ton of time to talk about it, and so I'm going to try to zip through it and then hopefully we have some time for questions. So, just a history of the farm. So our farm is called New City Farm. It's a non-profit. It's a part of a non-profit that does youth development work. We started in 2012 with the particular farm project, and then we grow on three acres. It's a social enterprise, so youth employment is the social side of the equation. And then it's run as a small business, that's the enterprise side of the equation. And just to give a sense of what the enterprise side is, like we gross about $80,000 a year in vegetable sales, and last year we did about $20,000 in value-added sales, so the pizza farm is kind of part of that. That's about 80% of our program costs, and then the other 20% is recoup through grants or donations, helping support the social side. We started doing value-added soups before pizzas actually in 2015, so our farm is actually located on the property of a church, and they had the three acres of land. They also had a kitchen that could get licensed as a commercial kitchen, so early on we realized what an asset that was to literally have 100 feet away from our field, a licensed commercial kitchen. So we started doing value-added soups so that when our CSA sharecroft customers come, they can buy a quart of prepared soup as part of their CSA, so $7 a quart. We put them in a little refrigerator at that CSA pickup. So that's our farm. I'm standing on the church's steeple right now. It's a flat steeple. I'm not like on a point or standing on top of a cross or anything like that. And so you can see now from the field, looking back to the church, and then you can see these are the exhaust fans to our kitchen. So really close to a commercial kitchen. So how can we utilize both this neighborhood asset of field space, but then also that kitchen? So these are some of our youths doing everything from the other nice thing about this. We're doing a youth employment program. We were doing four years of growing food with kids. That's kind of fun, but not that fun. Eating soup is a little bit more fun. Eating pizza is sweet, right? So I don't have to push a high school student to eat a pizza, but I got to push them a little bit to hand weed carrots for like three hours, right? And the other thing is you will weed carrots better after you've eaten them, not the other way around. And so a lot of our youths were working in our program and not necessarily getting to eat the fruits of the labor, so to speak. So then once we started making pizzas and soups, we do a staff meal every Wednesday, and whatever soups the team has made that week, that's what the whole group eats. And so two students are working in the kitchen, 10 or maybe out in the field, but they can say like, oh, Aiden, good job on the carrot, ginger, soup, or whatever. So again, we got started in value, not only because of the kitchen, but also because of Joel, who's not here with us, but he's my coworker. And he really had a lot more cooking experience than farming experience, but he had both, which is also important. And so he had about 10 to 15 years in a professional kitchen environment. And so he knew a lot about food prep. Food production is just like farming. It's got a big learning curve to it. You can have a garden, but farming is making money while you're gardening. So you've got to have the right systems, and cooking is the same way, just because you can make really good soup at home, it might have taken you two hours. So you've got to make really good soup, and you've got to do it at a pace. So just kind of keep that in mind. Joel brought all those skills with us. He also had a lot of food safety knowledge through that, and that becomes extremely important. And you're thinking about doing value-added production. There's no better way to put your farm completely out of business than to give somebody a coli. So when you get to the value-added level, there's no intermediary. So you've got to be really on point with that. So I always tell Joel, don't kill anybody. Because if he kills anybody, not only is that bad, but the farm is out of business, too. The whole user point, it's just all of it goes. And I know of a farm that they were doing dairy, and then they were making cheese on site, and they did have any coli issue with their cheese. Both businesses go bankrupt at the same time. So be on guard. So as I said, we started with soup, and then we did some condiments as well. We got started with pizzas because of Heather. I should know her last name. She just did a farmer-to-farmer podcast about two months ago. So definitely check that out. Curtis Milsamp, who also was quoted in the thing, he's doing pizza farming. So there's a couple farmer-to-farmer podcasts. But there was an NPR article. I was driving to the co-op to get cover crop seed. And they're talking to this woman, Heather, in Madison, Wisconsin, not in Madison, Wisconsin, about an hour out of Madison. And she's just talking about making pizza. And all these people showing up eating pizza. And I got back, and I told Joel, we're making a pizza. And he's like, of course we are. And so that's where the idea came from. So I'm not original in that. And Wisconsin is really pretty far ahead with this whole pizza farming concept.  Check out, and you can go online. And it's like this new agritourism thing that's bringing people out to their farms. And her farm is beautiful. It's in this beautiful valley. She's got playground equipment around. Wisconsin liquor laws are awesome. So she's got a bar. And she's got a band playing. And people wait about an hour and a half for a pizza. She's doing like 300 pizzas a night. And I got three small kids. So I'm seeing myself watching my kids while I'm drinking my beer. And they're not asking me anything. I'm just drinking my beer and eating pizza. And that's like, there's something there, right? So we got a farmer rancher grant. This is kind of how we used it to go out to Wisconsin for a lot of time. Some of our staff time. We did some pizza demos for the farmers in our area. And then we also did a workshop when we built our own pizza oven. And this is what we were trying to do. Compare profitability value added soups and pizzas. We want to know which one had a better margin. We also were trying to think about which one uses seconds better. That's a huge draw, right? So one of the things that Joel really likes to do is make sweet potato sauce for our pizzas. A, because most people have never experienced that. And B, because it's amazing. And then C, it's because every year 10% of our sweet potatoes have a mouse bite on the top of them. And so he just chops that part off. He gives it a good healthy inch away. But then all that becomes sweet potato sauce. So tomatoes are also notorious for giving you seconds quality. We use, you can't use slicers to make pizza sauce.   Soups will gobble up seconds a lot more. But that's kind of what we were trying to think about. We were also trying to think about how it might increase our CSA share. So, and Michigan, we are definitely post peak CSA shares. And it seems like that's a pretty common theme as I go around. So how do you differentiate yourself in the market? And we do our pizza sales at the same time as our CSA pickup. And there's some pros and cons of that. The cons are you got a lot of stuff going on at the same time. The positive is when your customer is coming to pick up, you've just changed like a weekly chore into like the weekly highlight. And so the three kids that are in the car after school are now eating pizza and they're really happy. And it's not like, oh, we had to do that right between. It's a fun thing now. And it's not this another annoying thing that I have to do between all the other things that I have to do. This is Heather's farm. She had a pizza oven outside initially. And then as she wanted to extend her pizza nights, she actually built, this is her initial kitchen on the right, and then this room was off of that. And that oven was originally outdoors. And then as her business has really grown, she then added, she expanded the kitchen because she needed more space. And then she actually moved the oven indoors because she's doing pizza nights in October now and it's Wisconsin.  You don't want to be slinging pizza in like sleet and all this kind of thing. She's also got a nice barn and sleeting area. So that's her bar. I like beer if you don't notice. This is her menu. So she's doing 16 inch pizzas. We do 12, but just to give a sense, she's just $20 plus for a pizza. And that's the right price because these are all local ingredients. Good flowers, good meats. She raises meat on her farm. We also visited a smaller community farm in Wisconsin. They built this pizza oven. So they're also just trying to scope out different pizza oven ideas. Whereas we were talking in that, what we had just seen Heather's farm and what was so interesting about hers is like, what we learned the most that we needed to be thinking about was crowd management. Because you're gonna have a lot of people coming to your farm. And I'll talk about that a little bit later, but it was like, how are you gonna manage this and just flow. And then this oven, it was cool, but we're like, where's your commercial kitchen? And it was like five blocks away. And so we were kind of like scratching our head. We're just saw Heather have to like, add another kitchen space, you know, and think really carefully about that. And we were kind of like, that's cool. Like that would be cool to do community garden things, but to do pizza oven night events like that, you just have to be thinking about that. And we were kind of like, yeah, we don't know. So anyway, and that's kind of their space. They're thinking about eating in. So that Milstaff farm, this is, I made a lot of this presentation last year, but episode one of one of the farm of Pompecas has some great stuff about how they do pizza. They also do it in a way that they've kind of bypassed their food inspector. They've did it while talking to them, which is important. Don't just like, think you're gonna bypass the food inspector. That's not a good idea. But they kind of did like a club thing, kind of like a CSA thing. And it sounded to me a lot of the ways like when a dairy farmer sells raw milk. It's kind of like they were doing the same thing with pizza. So you don't necessarily maybe have to have a commercial kitchen, but you got to talk to your food inspector or whoever before you make that decision. I'm not gonna talk about how we build our pizza oven because I don't have enough time to. I'll just flip through a lot of pictures to let you know that it's a pretty intensive project. I have a construction background. I actually used to work in Stucco before farming and then did home renovations for a while. So this was like my dream, like this was just fun for me, but it took a frickin' lot of time. But there's a YouTube video that we did do. It's on YouTube and there's a couple other videos on it. It's on our Facebook page. I'll have my contact information or if you go to newcityfarm.org, my contact's on there. You can email me if you have trouble finding it. This is just to say, there are some different options in building that. So you can build these ovens. So you're cooking pizza at 800 degrees. The oven is gonna be up at 1,000 degrees. So you're not gonna play around with this. Like things like blow up at that temperature. Molecules really like to expand at 1,000 degrees. So things will crack and you gotta know what you're doing. So you can build it. You can get a kit from FornoBravo.com.  You can get a kit with just, you're not gonna go to Home Depot and buy bricks from Home Depot. You gotta have fire rated bricks for that. You gotta have insulation around it that's all fire rated up to 2,000 degrees. We got like a pallet with all those ingredients in it for $2,000 for our oven which is I think a 36 inch diameter in the center. And then we built it brick by brick. Heather built hers with a module which is that middle picture. And I think that's around about $8,000 and they ship you like six really large plates and it's a lot easier to put it together. But your price obviously just went up quite a bit. Or we have a wood fire place that just opened up in us and like one of those downtown market things and you can get the whole thing shipped to your location too. And that's like about 12 grand depending on the shipping but they're gonna just drop the whole thing right there and you don't have anything to worry about they know what they're doing. So yeah, I'm gonna flip through. We're up in Michigan so we got a good frost line. You don't want this thing to start leaning over after a little bit. It's also heavy, it's probably like 4 or 5,000 pounds literally.  And then we needed a space. So Heather has this great advantage of being like in a rural area and this beautiful valley. And so her draw is like we're having pizza in this beautiful valley. We're an urban farm so we don't have that same dynamic so we wanted to create like a nice outdoor eating space. And so this last spring, the spring before we built the pizza oven and it kind of went into August and that's why it took forever because it's like build it for six hours and then go weed and then back and forth. And then this spring we built an outdoor patio. That's our commercial kitchen. We had to do a pass-through window after talking to our food inspector and come up with a plan for how that's gonna be doing. The other thing I was gonna say, talk to your food inspector or maybe talk to your extension agent first and who should I talk to? Because sometimes you can get certified through like we call it MDAR, the Michigan Department of Agriculture but I guess you're, how do you guys pronounce it in Indiana? Who is your, what's the, how do you say it? Okay. That's harder to say. But you wanna talk to them because sometimes those that food safety stuff can come through them. But we actually, and there's different ways to do it but we already had a food inspector because we were licensed commercial so we started talking to them and what do you need, what do you want? And generally they're pretty helpful. The other thing you wanna talk to particularly if you're in an urban space is your zoning department because that also matters. And so for example, we really lucked out we could not cook within 200 feet of a residential property and where we initially wanted to put our oven was like 190 feet away from residential property. And then we were able to just move it 10 feet and put it, and that thing is a tank. If I would have built that thing, like I was saying when Korea does their thing, I'm gonna crawl into the oven. And so I was like, we couldn't move it. So you just wanna talk to zoning and figure out like what do they want because if somebody complains they can shut you down or they can be like, oh, you have to move that 10 feet.  So think about that. This is what the spinach base look like. And then this is people eating pizza. And these are students loving to make pizza and loving to sell people pizza. And these are some of our pizzas. And so then I always like to have some kind of numbers behind things because you go to presentations and you hear these amazing things that people are doing and then how are they paying for it is what I always wanna know. But these are some of the things that we do in terms of soups and in terms of different kinds of pizzas we make. But then here's some of the numbers and these are just based on 2016. That was when our project was funded. So we grew quite a bit from that year to this year. But really what we were trying to tease out of these numbers is what is our margin? Like revenue doesn't really matter. I don't know why farms like love to talk around revenue so much. I'm more interested in what your hourly average wage is. That's the number we're always trying to measure. And by the way, I've done that with seven farmers and the average of the seven farmers was $6.66 an hour, which is a demonic number for one, which is a problem. But also it's just a problem. So like what Chris was saying, if the numbers don't work you're gonna be stressed out all the time. And that's part of what's leading us to think through this as well. So when we're doing soups, we're making $32 an hour. So I was making 6.66 and now I'm making 32. When I'm doing pizzas I'm doing $22 an hour. And then when I'm doing pickles and condiments I'm only doing 16. Now that's not a true comparable because these are just, these are variable costs. They don't have our fixed costs in it. So fixed costs would be all of our infrastructural costs that are just gonna hang over. This is just us like, what are the ingredients you put in the pizza and how many hours did it take you to prep that pizza, make that pizza, sell that pizza or do that with the soup. And so when I did the 6.66 number that was everything. That was fixed and variable. So it's not a true comparable in that regard. But it would be hard for us to tease all that out. We have a free kitchen. So that's a big thing to think about. If you were gonna, so we thought, we have some incubator kitchens by us. So we were particularly thinking about soup doing incubator kitchens. Incubator kitchens by us are about $20 an hour. And so if you went in there with just yourself and you were making soups in an incubator kitchen you would still be making $12 an hour. So that's like fixed and variable at that point.  But then if you put two workers in there or three workers and you increase, you're maximizing that hourly thing, your hourly wage will go up. If you're doing condiments, we found you're actually losing money by the time you do it you would lose about $3 an hour. So you might feel good about like not throwing away all those cucumbers and not pickling like, okay, I all pickled them and everything but people don't really want to pay that much unless it's like McClure's pickles or whatever. But even that, I'm sure they have like some pretty significant processing facilities that allow them to do that at scale and at a pace that's efficient. If you're just doing it kind of in an incubator kitchen I think you're gonna be hard pressed to find that you're gonna make a good hourly wage. Is this a good use for seconds? So this was another question that we were trying to ask. So soups down here, percentage of vegetables that we're using that are second. So soups just kill it. I mean, soups are like pretty much all vegetables and there's a lot of seconds that you can easily use in soups whereas pizzas, A, you're buying in dough and oil which we don't do, we also don't do animals. So we're buying in, we do do meats on our pizza and we have a good local butcher that we work with. So we're paying for quite a bit of the ingredients off and then we're making our own sauce and we're doing all the vegetable toppings. So we don't use as much seconds with that and you can use quite a bit of seconds like a lot of times you get a broccoli head that will go wonky or something like that. Joel loves broccoli seasoning he's making broccoli cheddar soup all the time. I love Joel for that because then all of the broccoli that I should have harvested yesterday aren't just going and getting composted, we're using that stuff and then he's actually making more money when we do that than if I sell it. So I shouldn't even draw the regular broccoli I guess. I don't know, he's kind of mad at me but. So here's some things that we thought that you might want to think about. Is this the right thing to do? So kind of getting back to the capital investments we had some assets in place. So if you didn't have those assets in place and you had to put in a commercial kitchen you need to run the numbers to make sense of if that's a good decision for you. One idea I have had, because most of that's the biggest pushback I get when I talk about this is I think if you are a farm like and you're in that moment where you're gonna like up the pack and wash station I think there's some interesting space to overlay the two together. A lot of the things you need for a commercial kitchen like a washable wall, a triple sink, a hand washing station. These are all things that a good pack and wash station need. So maybe you design your, I just saw, who did the talk yesterday on the wash and pack station? What's that? Yes, he was great. His wash and pack station, one of the things he designed was like hey this was an urban farm all their sinks and stuff could move in and out of that wash and pack. So I'd be really curious to build a wash and pack with having that in mind. Maybe we're using this as a kitchen to make pizzas two days out of the week and we're using it at our wash pack when we're going to farmers markets. I don't know, it's just an idea or I thought I don't have to figure it out because I have one, but it's an idea. But you do want to know what your fixed costs are like and then depreciate that over time and be like okay, that patio was a $45,000 investment for us. So that's going to take a while to pay that back. So think about that. The bigger thing to me is do you have the skills for commercial kitchen cooking? And so don't think it's not a skill set. Like I think times people don't think farmers really have a skill set, but then when you do it you're like this is really hard to figure out how to weed effectively and efficiently and to make money doing it. I think cooking when I watch Joel is completely the same. So if you don't have, I think in the same way you would expect somebody to run a farm that they've worked out a farm for a couple of years. I think you need to work in a commercial kitchen for a couple of years if you're going to get into it. You can do it the hard way too. I never actually worked on a farm before I started farming and it took me a lot longer to make something decent of a living. And then you got to make sure you insured food safety. I think I scared you about that enough earlier. But then some value that you also can think about. So you have an increased exposure to potential customers. So we had a lot of late CSA signups this year because they were eating pizza. So the pizza cafe is open to the general public. They're coming for pizza on a Thursday. They get to see the CSA pick up at the same time. So we drew in a lot more CSA customers. Probably another five or 10 CSA customers in June when the cafe opened. We only run the cafe from June to August when all of the students are out of school. The CSA we run from May to the third week of December. But in that moment when the cafe was open we saw a lot more people interested in it. And then we haven't measured because it was between this year and next like what our CSA retention numbers are if they change because now people are having a more satisfactory experience with their CSA. I think the other thing we've always struggled with with CSA is we've never done community events well. You know like some CSAs are like really great at like loving their customers and bringing them on farm. I just have, we've just never have really done that well and this became like a win-win for that because now they're hanging out on the farm and we're making money at the same time. So we're being fairly compensated for all that effort because it's a lot of effort to put on an event like that. And it doesn't feel weird for somebody to be paying for a pizza at it. You know, nobody was complaining about that. And then it was fun for us. Like it was fun for me to hang out in the cafe and then I could eat pizza with a customer that's been with us for three or four years. And when they're just going through a CSA line you just don't have the same experience. So I really enjoyed that part of it. And then for us it was a great educational thing to bring that our employees all send go from the full circle right all the way from seed to pizza. And then existential questions.  Because like Heather's business it's a pizza farm more than it's a farm from when I went and experienced it and kind of as I heard her talk about it at the podcast more of her revenue I think is coming from pizza than it is from her two acres of vegetable growing. And I think when she was talking to Chris Blanchard she was, I think they even stopped CSA. So they've kind of almost gone through the entire circle and just realized we're gonna just grow on these two acres for our pizza nights. Which also kind of gets at the other panel's thing when it's like how do we do the farm to table thing well and truthfully? Like let's just do it all ourselves because then you don't need to deal with an annoying chef who's like greenwashing you. You're doing it all yourself. And then the other thing I kind of touched on a little bit will you enjoy weekly crowds at your farm? If you're gonna have 200 people show up what are they gonna do for the two hours? Heather was talking about people falling into her fire that she had and kids trampling on her vegetables because there's a whole bunch of kids running around. Where are they all gonna park? Where are they all gonna go to the bathroom? You gotta think about all those things. So crowd management, where are, how are you gonna move all those people? Think about lunch and the really long line that we had at lunch today and how that's stressful. And you saw the caterers being really stressed and the people in line really stressed. You can only make like four pizzas at a time. They cook in three minutes, but you can't make 200 pizzas all at the same time. So Heather has lines like we're getting them out pretty quick. Like in an hour, we didn't have anybody wait more than 15 minutes. We also didn't advertise to anybody. So to put in perspective this year, we did like of this last year, we did 10 nights and we averaged about $2,000 in sales every time we did it. It was on a Thursday from 1230 to six. So it wasn't even on any kind of prime time hours. And we advertised to no one. I was like, don't tell anybody about this because Heather freaked me out and I didn't want all those people coming. And actually that was the right crowd for us. Like that was like perfect. We got a good rush at lunchtime when our first CSA customers are coming and we had a bigger rush at dinnertime. But we could get them a pizza in 15 minutes but we didn't have the band playing for an hour. And that's why Heather had to kind of figure out what are you gonna, if they're gonna wait for an hour, what are they gonna do? Or maybe you have to have two ovens or three ovens. So kind of thinking about all that. So I think that's all I got. Those are my contacts, my email and those are our website and our Facebook page. And yeah, shoot with question. So there are mobile wood-fired pizza ovens too. And something to think about too is maybe your farm doesn't even wanna get into this but maybe you do wanna have four pizza nights at your farm. And I think a good option is talking to the mobile wood-fired pizza oven guy and say, hey, will you use my toppings? We'll collaborate in advertising this thing.  And now you don't need to know how to do pizza. You don't need to have any infrastructure in place and you have 10 really cool events at your farm. You get some of those other tendential benefits of like CSA customers are more excited and more interested. So that would be where I might think about starting. Yeah. Oh, so did all the ingredients for the soups come from the farm? So no, like that percentage is like 90, like over 90% did but we don't make cheese. So if he's doing a broccoli. So you just labeled that like for the transparency stuff. How do you communicate that? Yeah, so what we do is it's a quart jar and it has a little card on it with a label on it. Like a glass ball jar? Glass ball jar with a little string with a little label on it with our logo on it and a broccoli cheddar soup. It has our expiration date on it. So when he makes the soups, what he does is he hot packs them and they seal and they'll keep very long because they seal we only let people keep them for two weeks but we actually used to have an expiration date of a month and we bumped that back to two weeks now but they have to be refrigerated the whole time. If you know anything about canning, like it doesn't do anything for botulism but it will preserve it quite well but it has to be, it has to stay below that temp the entire time and you can't, you don't get botulism. Yeah, so it's just like keep refrigerated used by two weeks past when he made it and then it will have all of our ingredients and there'll be an asterisk and every one of them that are off of the farm and then we only use organic ingredients when they're sourced from off. And do you have to include nutritional information like you see in the store? We originally did that and the food sector actually said we don't need to do that. So we did originally do that and when we talked to him about it because of the limit, like I think if we were going into a retail establishment with it, then he was gonna want that but when we described the context by which we were selling it, he didn't require that. When did your customers bring those drugs back? Yeah, they love bringing them back.  We're still buying jars for sure but like, I bet you about 10% of the jars we use we're actually purchasing and then the rest come in and then really what we're doing is kind of buying some and then there's just ways where they kind of forget and if we send out an email to the shareholders or put up a sign at the CSA pickup, hey, we like, love to have your jars back then people will tend to remember and they'll bring like 15 back but a lot of times I think it's just, it's something people forget to do. So we're in an urban neighborhood so it's very easy to get employees because there's a lot of kids that want to work and they don't want to work at McDonald's which is their next best alternative and so yeah, so we've actually never advertised a high school job position. It's generally word of mouth just through our former high school employees and stuff and as we've built relationships with some of the school we let them know of the opportunity and sometimes a counselor will kind of identify some student and send them our way but mainly it's from the students talking to their friends. Yep, so first year students come as an intern and so they get $1,000 stipend for the summer then second year students get $9 an hour and third year students get $10 an hour.


logy of sort of testing and experimentation and falsification of hypotheses and communication with other experts who are doing similar work in the field rather than a methodology of, you know, just make some inferences and then, you know, claim that that's absolutely true. So until such time as I've been presented with better evidence, I'm going to go with the theory of evolution, which at this stage is one of the pillars of modern medicine. Thank you. Thank you, muted, Amy. And thank you all for watching. I always do that. It's the double tap. Thank you to Amy and Church of Entropy and, of course, James for letting me on here. It's been an absolute blast. And I believe both Amy, you and Jen are muted. No, it's the Zoom meeting. It is. As long as she all transitions. I can't hear Amy. I'm not muted right now. I just checked. No, it's the Zoom call. I can see the mute is on the Zoom call. Should I stop the screen? There you go. Yes. Thank you so very much, Mark, as the screen share. I love. Don't even worry about the screen share. Seriously, don't even worry about it. It looks like an absolute train wreck, but I honestly love it. And don't even worry about changing it to something that looks good. So if anyone wants to see the actual model, you can see it not looking like a train wreck on my channel after. But yeah, it's some evidence requires a certain standard of axioms. You have to accept the axioms to get proof, quote unquote, or evidence in support of more accurately, right? So the theory goes beyond that because it's telling us not just what's happening, but also how to frame what's happening.  These metaphysical assertions and decisions are. The ground of proof. So you're asking for proof. Like you're basically asking why you can't drive the engine of a car. You can't drive the engine of a car mark because you drive a car, but you need an engine in a car. Don't you? To drive it. So there's never going to be proof on that level. The argument is basically this is a more efficient means of getting to what you could call proof because it accords with what's been used traditionally. And I'm actually considering to keep these things as simple as possible. Informationally, though, there is this big initial. Learning curve. Once you get it, it's there and you can just draw it at any time you want. So hopefully this diagram is a little bit of an. Insight as to how the model works. Yes, you have to look at reality. But once you transcend reality, you can find the. What are called margas of things, the root. And the openings and everything basically functions on that basic. Premise, which is why on the bottom right talks about the way of mind. So we can go into that more in the after show. And I think do have it will be joining. So please check us out. Church of entropy. Thanks again. Sorry for the long answer. Thank you so very much, Jen. And that's right. You can both check out the after show either on Jen's channel or at the modern day debate discord. But I do want to thank you all for joining us out here on modern day debate. Katz and all we are a neutral platform welcoming everybody from all walks of life. If you're looking for even more juicy debates, don't forget to like and subscribe, including tonight's debate on evolution on trial. I want to thank both of our interlocutors, Jen and Mark for joining us tonight. And if you liked anything that either of our guests said tonight, you can see their links in the description or the chat. Go out and check that out, ladies and gentlemen. With that, I am Amy Newman with modern day debate, and we hope you continue having great conversations, discussions, and debates. Good night, everybody. Good night. Thanks for having me. Are we done? Are we up? Sorry, I can't hear you. I can't hear you if you're talking.