 Okay, great. Well, thank you for coming. This is the Ethics and Research and Biotechnology Consortium series. As you know, this is a series that brings bioethics and research ethics to some of the latest research in biotechnology and bioengineering. Let me just go over some ground rules for those of you who may be joining us for the first time. We welcome questions from the audience during the talk and to do that, please enter your questions into the Q&A box found at the bottom of your screen. Don't use the chat box unless you need some technical assistance and the chat will go directly to the panelists and to the administrators. At the end of the presentation, we will have time for a Q&A with the audience and I will be moderating that. Look for upcoming events for next year at the website bioethics.hms.harvard.edu. And let me go ahead and get started introducing our speaker for today. So Dr. Julie Kim is the CZ Y-Hung Professor of Aesthetics and Gynecology at Northwestern University's Fiennberg School of Medicine. She is also Northwestern's co-director of the Center for Reproductive Science. And she received her PhD in Cellular and Molecular Biology at University Laval in Quebec. Her research focuses on understanding the development and growth of diseases that affect the uterus. She's helped develop physiological models of the female reproductive tract, which can be used to interrogate research questions surrounding fertility and disease and for testing new drugs. So she's here today to talk with us about the development and use of some of these modeling systems. The title of today's session is FEMA TAR and Other Bioengineered Models of the Female Reproductive System. Dr. Kim, the floor is yours. Welcome. Thank you. I will just share my slides now. Okay, I hope everybody can see that. Great. I really appreciate this invitation to talk to you about our research. And I think this is a great platform for all of us to share ideas, not just from the biological perspective, but really from the ethical perspective as well as reproduction is highly, a subject that is highly discussable or conversable in the ethics field. This is the slides are changing just by themselves. So, all right, I have nothing to disclose before we start. And so because this is a more broad audience and wanted to give you a good background on the female reproductive tract, I know you've heard a lot about some of the other fields of biology. But here we have the female reproductive tract that consists of multiple different units that makes up the whole tract. And so, for example, we have the ovary where we have follicles and oocytes that develop and are eventually ovulated. And once ovulation occurs, we have an organ called the corpus luteum that then secrete different kinds of hormones. Once the egg is released, it goes through the fallopian tube. I'm sorry. It goes through the fallopian tube where if there is an egg and sperm present, that's where the egg is fertilized in the fallopian tube. And then we have the endometrium or the body of the uterus and the lining of the uterus is the endometrium, where eventually the embryo will attach, implant and start to grow. We have also the cervix, which is the actual bottom part of the reproductive tract as well as the vagina. And so those are the units of the female reproductive tract that we should keep in mind. And really the goal or the function of the female reproductive tract is to be able to carry a fetus to term so that we can eventually have live birth at the end of approximately nine and a half months. And that is why it is such an important organ to have because it is essential for the propagation of our species. And so this is a little bit more detailed look of what the female reproductive tract does. As you can see here in the ovary, there are follicles which are made up of the egg, the oocyte, surrounded by support cells, usually granulosa and fika cells. And this is the unit that releases hormones such as estrogen and progesterone. So once the hormones are released and the egg is released, the hormones are shared throughout the reproductive tract to make changes of the reproductive tract tissues, the fallopian tube, the endometrium, the cervix, etc. And so the estrogen, which is predominant hormone during the follicular phase that's before ovulation occurs, it provides proliferation or growth capabilities of some of these tissues, and especially for the endometrium. Once there is ovulation, then the corpus luteum, as I mentioned, forms, progesterone is released, and progesterone then does even more things that prepares that uterine lining for implantation occur. It's a very specific window of time where an embryo can implant. And that means that the endometrium has to be receptive. It has to be perfect for that embryo to attach and implant and establish pregnancy. And so this is a very orchestrated and very controlled process that occurs every 28 days. Now, if there is no pregnancy, there is no embryo present, then of course that lining that's prepared is shed when the hormones levels drop. And so this is a more graphical presentation of what happens is when you see the follicles on top there growing and then ovulating in the corpus luteum forming those are the, that is the source of estrogen and progesterone. And in order for those follicles to grow, we need signals from the brain. These are hormones that are released by the pituitary gland, luteinizing hormone, as well as follicle stimulating hormone. This is what controls the growth of the follicles. And like I said, there's estrogen and progesterone being released. But what we can appreciate here is the fluctuation in the levels that change over the course of time. We have a peak of estradiol that's formed during the luteal, at the follicular phase, and then the rise of progesterone that occurs in the luteal phase. And that those fluctuations of hormones are what changes that endometrial lining. And this fluctuation, this partnership that occurs between the estrogen and progesterone are key for that regulated and very controlled growth of that endometrium. And so we see that it's a complicated process. There's a lot of things going on within the one menstrual cycle. When there is no pregnancy, like I said, the lining sheds, you get menses, and then the cycle starts all over again. And so we know this because of the research that has been done in humans as well as mouse models, but there are obvious limitations in the kinds of research that we can do with female reproductive tract. Mostly because women are born or girls are born with a reserve of follicles. That means there's only a set number of follicles that can ovulate. We can't make more yet. And once that reserve is used up, then there is no more. And that's hence that's what happens during menopause. And so because there is limited reserve, there is very little that we should be doing in order to insult or put these follicle polls at risk. Research is also limited because there's vast species differences in the reproductive tract. We've learned a lot using mouse models and primate models. But there are limitations because not all of the processes are identical. For example, the placentation of humans is very distinct from the rest of the species. What happens in humans is that the trophoblast or the embryonic tissues, they infiltrate or they invade very deeply and they just tap into the maternal vessels so that they're sharing of blood very early on. A lot of the species don't have that. Most of them are superficial implantation. And so knowing those species differences also. We can use that information to our advantage as well. And I'll be showing you later on how we can use differences, but then understanding the biology of what drives the human reproductive processes. We can then use those signals to make other species behave in a way that the human tissues behave and I'll go into that more in a bit detail later. Sitting hormones, especially female hormones, estrogen and progesterone, it's quite complicated because we're talking about fluctuations as well as partnerships between estrogen and progesterone. That's really hard to mimic in vitro. Also hormones do act all over the body. They don't act just on the reproductive tract, but they do affect almost every tissue in the body. And their actions are very context dependent, depending on the cell type, the tissue, how much there is, how long it's been exposed. And so every context is different. And with that said, because of this, let's see, yes. A lot of the preclinical and clinical research has been done primarily in males. I mean, it's difficult enough to have a clinical trial going and to getting data at the end. But adding people thought that adding in women to the trials would introduce a lot of complications, a lot of variability, because of the hormone variability that I just mentioned. But then the result of that, of having a lot of these trials focused on males, the result of that is that we don't really know what the side effects for women are going to be until the drug is already on the market. And that is what has been observed for many drugs that have come out. So eight out of 10 drugs have been pulled from the market because they had serious side effects in more in women than in men. And some of those are listed on the right there. Those drugs that have been pulled. Now, for example, if we take stentons and many, many people take stentons for cholesterol, we do see a lot of differences in terms of the side effects for males versus on the right versus females and on the left. And so these six differences, we cannot ignore them. They do exist. But how do you study that? How do you test drugs without putting that reproductive tract in risk or the fertility function or compromising the fertility function of women. And so this is the big gap in knowledge that we have right now. And because research is limited, and then a lot of drugs cannot be tested directly in women that can affect the reproductive tract, we don't know a lot. And, you know, it's amazing to know that a lot of the diseases that affect the female reproductive tract as listed here, there is not a great therapy for any of them other than surgery. So infertility, you know, it can stem from bad oocytes or follicles. You know, you can get ectopic pregnancies because the fallopian tube is not able to brush the embryo into the uterus. And there are a lot of endometrial disorders as well, including endometriosis, endometrial cancer. Even the muscle layer of the uterus can form tumors, such as fibroids, then there is cancer of the cervix cancer of the ovary that can occur. There's premature ovarian failure on young women and not having those, those eggs that are reserved. It's not usable. And then there's a polycystic ovarian syndrome, which I will describe a little later, that affects not just the ovaries, but a lot of other organs in the body. And so I guess the take home message for this is that there are still pathologies that affect the female reproductive tract, they're called women's diseases. There's really no good therapies right now to eradicate or to cure any of these. And this is because it's really hard to study this outside of a woman's body. And so that was our research goal is to develop a physiomimetic or something that we can test in the lab outside of a woman's body, the female reproductive tract to ultimately use as a tool to better screen compounds that come into market systems, but also to better understand the biology of reproductive tract, and then ultimately understand why diseases happen, and what we can do to combat those diseases. And before I move on, I just want to acknowledge the team that was involved in the initial building of this reproductive tract in vitro. It takes a village, honestly. And this was really driven by Teresa Woodruff, who was the PI of this program, about 10 years ago. And then we had collaborators join over debt, we focus on the fallopian tube. Myself, I was in charge of building the uterus, a spiro and Thomas hope the cervix. And then we worked with engineers john competitor, and Jeff Bernstein from Draper labs. And so again, I just want to acknowledge this whole team, and I'm just speaking on behalf of everyone on this project. And so about 10 years ago, the Tisha chip consortium was introduced or was developed. And this was basically NIH and dark by getting together and offering funds to create physio mimetics. And so organs on a chip microfluid microfluid physiological systems, where organs can be represented outside of the body, and placed in cultural systems, where it can mimic potential blood flow and more physiological components, so that we can eventually test drugs and compounds in a meaningful way. So this would be a huge a significant advance to what is already available. So we were able to obtain some of the be involved in the Tisha chip consortium and build even tar, which somebody very cleverly coined the mother of micro physiological systems. And basically this would consist of a system a microfluidic plate, where we can house different compartments or units of the female reproductive tract, including the ovary uterus cervix fallopian to. And then we put in a liver in there, in order to understand what would happen if a liver was present that with the refame of a reproductive tract, in hopes that we can add in compounds where the liver can metabolize these compounds. As I said, the collaboration was built between Draper labs and our PIs at Northwestern and UIC. These are the various people that were in charge of building a specific their specific organ of interest in vitro, and then Draper labs. Basically they built this microfluidic system, it so that these these specific units could communicate with each other, and that that would be a continuous flow of media, kind of mimicking the blood. And so this was the platform they built and designed for us. It's basically a plug and play kind of unit where each of these compartments you can take, add in or take out. And once they're added in, they are all connected through microfluidic channels, and so they would share media in any specific direction you tell it to. But the first, the criteria for building such a system would be that it could be sterilized because we all know in vitro cultures in vitro, it cannot be toxic to the cells or follicles, depending on the materials they're made of. Sometimes they do leach toxins, and that would be detrimental to any of the cell types that we use. And this is something that's unique to our system is that they cannot bind hormones. So a lot of the fluidics platforms out there are made of substances such as PDMS, which absorb hormones absorb hydrophilic compounds. So that the actual cells don't see them as in the same concentrations you think they're supposed to see so that would not be conducive to our system. And of course these pumping systems would be computer controlled and it would be customized to whatever we think the flow rate should be. And so this was built. And we also needed, because we're talking about the human menstrual cycle and I give you some background on that. We also needed a source of hormones that would not only provide them in a certain level but they would be fluctuating over a period of 28 days. And so this was a challenge. And we, we thought the only thing that can really do this is an ovary. And so this is where Teresa did her magic in that we used mouse ovaries, instead of the human ovaries to to provide that source of ovarian hormones, including estrogen and progesterone. So this was kind of tricky, because the mouse ovary or the mouse their astro cycle is only four days long, whereas the human menstrual cycle is 28 days. But as I was mentioning before, knowing what drives hormone production. And in what context, she was able to treat these mouse ovaries with LH and FSH, which are the pituitary gland hormones that then act on the ovaries to grow the follicles and mature them. So she was able to add in FSH and LH in the program, or like a human menstrual cycle. And so she would add an FSH over a period of 14 days, and assuming that the mouse ovaries would respond to that. And she would cause them to ovulate by adding a high dose of LH or the, what we use in the lab as HCG, which is an LH mimic, and then continue that on so that progesterone can be produced. The reason why mouse ovaries are used here is that there is not many human ovaries that we can use from the clinic, nor should we because we are looking for ovaries from a fertile woman, so premenopausal. And these tissues are very hard to come by. Plus, as I mentioned, women have a dictated reserve of follicles that you don't want to mess up with or put anything at risk of fertility. So that is an area where I guess the ethics committee can be really discussed and converse about this that, you know, the whole, is the human ovary necessary for us to do human studies in the reproductive tract. And so that is the reason why mouse ovaries are used. Julie, I have a quick question for you. Is there any interest by your group or other groups in the future to maybe use human ovarian tissue from younger persons pre pre cancer treatment to hopefully, you know, provide a means for them to have mature oocytes later. Absolutely. Right. And so that's where the, so Teresa would have also formed this consortium called uncle fertility, where they do just that. They are interested in preserving some of that ovarian tissue, so that younger women or even girls can undergo chemotherapies or what have you. They are treated men's that usually harm the ovary. But once they take that reserve and preserve it, then they can retransplant those later on after all of the therapies are done, so that they are able to eventually can see. Wonderful. I'm sure we'll return to these issues during the Q&A. If any of you have questions for doctor camp is under that Q&A function. Thank you. So even though I said we don't have the ovarian tissues in the clinic to study. There are cases where there is healthy normal ovary that is removed in the context of, for example, prolapse where the whole uterus is coming out because the muscles aren't there to support it. And so there are cases like that. And we have researchers at Northwestern that are doing aging studies using human ovaries in this way. And so here is an example of what Teresa Woodruff's lab did using mouse ovaries. What, as you can see in the pictures, this is what a mouse ovary looks like. A mouse has multiple follicles in their ovary that can ovulate at the same time, whereas a human has usually one dominant follicle that eventually ovaries every cycle. But the mouse has multiple. As you can see that those circles in these pictures are follicles that are growing. And she adds an FSH over a period of time, the follicles grow to the point where they're growing to a certain size. She adds a bolus of LH and then the ovaries able to ovulate or that follicles are able to ovulate. And then because of these events, we have that increase in estrogen that occurs when the follicles are growing, and then the progesterone being released after ovulation when the corpus luteum has formed. And interestingly, using these mouse ovaries, she thought, well, what would happen if we continue to add LH, which is, we use HCG, like I said, it's like a chemical mimic of LH. And HCG, as we all know, is the pregnancy hormone, right? And that's one that signals to the ovary, hey, there is an embryo here. We need that ovarian support of progesterone. And that's exactly what happened when she added HCG over a prolonged period of time over the cycle. We got that high level of progesterone being maintained over the course of the cycle. So that was pretty cool. And that was really proof of concept that these mouse ovaries are indeed responding to these kinetics. And that was what was fueling or that was driving evatar. Those mouse ovarian hormones is what drove that hormone production. And then the rest of the downstream tissues, including the uterus, the cervix, the fallopian tube, was then able to respond to those ovarian hormones. Because this evatar allows communication of the media between the different units. And that's what we asked the engineers to do, is to provide channels to connect each of the compartments together, but also allow some recirculation to happen within each of the compartments so that there's a bit of mixing of the media. And so this is a schema of what was designed for the platform. We went from follicle to fallopian tube to the uterus to ecto cervix and then we had a liver. And then we had an acceptor well, which is basically media that's collected at the end of the one circuit. We had a donor well where it was fresh media where we added in the gonadotropes, FSH and LH exogenesis. Now it'd be great to add in a pituitary to be able to naturally release those, the LH and FSH. It's an incredibly complicated process. We're not there yet so we're adding in the LH and FSH on our own. So I just want to summarize what happened. What happened to the downstream tissues when we had all of these hormones there. The fallopian tube, we saw that it did provide that physiologic environment so that the cilia would be beating that epithelial cells proliferate. The endometrium also functioned and responded to the hormones in a physiologic way. There is a process called the sigilization that happens with estrogen progesterone. And that is what happened in the endometrium, the ecto cervix, that squamous layer grew, musons were secreted, etc. So what we learned from this study was that all of these tissues, when we study individually in each of our labs, they take a certain amount, there's certain type of media. But when they were all together, they all thrived in one media, which was amazing, which was suggesting that each of these tissues were able to secrete the factors that were necessarily to prolong the longevity of the tissues. And that's what we saw over the 28 days as well, that the cells were healthy, these tissue units were healthy. There was minimal damage at the end of 28 days, which is a very long time for something to be cultured in vitro. Julie, I have a quick question for you. So when this avatars system was first published and released, you know, into the public. Did you have any challenges in communicating what you did here to both media and to the public. I have to confess, I have a confession to make to you, Julie. When this first broke, I did an interview with NPR, and they had asked me a lot of questions, one of the questions they asked me was, is there anything to be concerned about for these kinds of ex vivo, you know, outside of body modeling systems of human reproduction. And I said the only thing that I could think of that might be of concern in the very distant future, this ex vivo embryo genesis or like ex vivo gestation. And I said, but we're not talking about that here. Now, when my comment was published in the article. This is the last little part about that's not what we're talking about here so I even found it challenging as a bioethicist to talk about this to a to a well respected media organization like NPR. What were your experiences like talking to me. Yeah, and then I would love to talk about this so here, great segue. And I did a ton of media attention, because basically this was the first reproductive tract ex vivo that was created. And so what I found was that people the reporters did contact us directly and did very nice interviews, and they were accurately accurately represented in the articles but then you do see a lot of these other news channels where they would just look at, not interview us but look at these other articles and then write their own things. And basically, I mean this is a pretty complicated process really to understand. But a lot of these, the higher news agencies were pretty accurate in describing our system so they called it the menstrual cyclone chip, you know, and it was, it was, it was pretty, it was pretty cool for everybody to to read about this but then you see the articles that label it as period in the Petri dish, or scientists have recreated a period. And that's the furthest things that we did, there was absolutely no bleeding or menstruation or blood vessels in this unit. And I found that, you know, I think that the potential inaccuracies that we sometimes see in the news media was really evident in this, in this study. I mean, you can't, like I said, it's a pretty complicated concept. When you talk about the menstrual cycle I don't think people know exactly what is involved in all of the different intricate steps. You talk when you talk about menstrual so you talk you think about periods, right, you talk about bleeding. And I think there was there was also one article, and I couldn't find it I was I was looking for online, because it was a very funny title it was like a vagina in a dish or something and it just made me laugh and I actually wrote back to the author and said this is inaccurate. This is not what we did. So, I hope that answered your question. Okay. So in summary of Evatar, we were able to reprogram mouse ovaries to secrete estrogen and progesterone in the pattern of a human menstrual cycle, which was a huge feat and a great thing to do. The downstream tissue is what we saw that they did respond to these hormones, and they actually really liked sharing media and peer factors with each other that increased their longevity. It made them more responsive to these hormones, unlike what we see when we're culturing them individually as separate units. Yeah, and so that's that's what we saw with Evatar. And so that was Evatar that was perfect concept. We showed that the female reproductive tract could be represented. There's just so much potential there that we where we can use this. So the next phase of our study was to model a disease using our microphobics. And so this is the second part to ship to. And really the goal was to understand the disease better because we are, we have multiple units looking at sharing media with each other that we can test drugs to see what happens when multiple units are there. We can potentially use it as personalized medicine, depending on what kind of cell types that we use. And then one of the things that we do in the lab that we couldn't do before is to understand if risk factors can directly act on tissue and damaged tissue. And so this is this opened up a whole new world of research of the female reproductive tract. The second part was divided into two parts. The first part was, we loved Evatar and all, but it was a very, very difficult to use, very expensive. You know, it wasn't as versatile as we wanted to because we were biologists and only engineers knew the language of how to run Evatar. And so we wanted to make a simpler system that was user friendly to biologists that were cheap, that was cheaper to use. So, but that provided the same concept, multiple units talking to each other. And so we have built that we call it lattice, we call it the daughter of Evatar. It's a second generation version. And I'll tell you all about that. The second part was choosing a disease to model. And there is a reproductive disease, at least from the ovarian perspective called polycystic ovarian syndrome. This is a real mysterious syndrome. It impacts a lot of women. And not only do these ovaries have multiple cysts like structures, but they don't ovulate as much and they release high testosterone. And so you see symptoms of high testosterone and PCOS women. But the other thing is, it's not just an ovarian disease. It impacts the liver, the pancreas, muscle. It involves multiple other organ systems because a lot of these women also have insulin resistance and obese, they are obese. They have high metabolic syndrome. I mean, there's a lot going on. And we don't really know what the cause of polycystic ovarian syndrome is, nor do we have anything an effective treatment for this disease. And so we thought we thought this would be a perfect disease for us to study in the multi organ fashion. So I just wanted to go over this picture just to show you what the evolution of lattice looked like. I mean, there were certain criteria that we had to meet. It had to be user friendly and to be familiar for biologists researchers, we all use those 24 well plates. And so we thought it would be great to mimic a 24 well plate. We had to make it from material that did not absorb hormones. And we had to make it from materials that were very cost effective. It had to be reliable, etc, etc. We did have a version before lattice that we use we thought maybe we can just 3D print everything because that would be really cheap. We did do that use materials that were conducive for 3D printing, but then found that these materials do leach toxins that are detrimental to the ovary and so that was scrapped. And so then we partnered up with engineers and other companies to outsource to build us lattice and what lattice is is basically a plate. And as you can see in the black plate with multiple wells that had microfluidic channels that connect all of the walls together. It has a pumping so and then the bottom part is the base station that controls pumping of media across these channels into these compartments. And so what the motors are and the sensors are, etc. And so very simple, relatively cost efficient, and it's pretty easy to use. And so, I want to show you this video, it was actually made from a very talented master's student at UICs, my biomedical visual program. And so what lattice does is it's a multi organ in vitro system to study dynamic interactions of up to eight unique organ cultures, like an interact with each other for extended period of time and here we chose 28 days which is a menstrual cycle. A liquid media supports nutrient exchange waste elimination and enables secreted factors to interact with different tissues via these micro scale channels and then lattice is connected to a computer, which controls the microfluidic actuation of the system, which provides precise media flow through each chamber, and there's a valve mechanism as you can see here so that precisely regulates the volume of the culture media that is transferred from one tissue well to another so it's a very simple system as you can see, it just rotates that media into one well to the other And so just a little bit more about polycystic ovarian syndrome, it's associated with hyper androgenism, ovulatory dysfunction and polycystic ovary. And again, I said, as I mentioned it's involved is associated with insulin resistance cardiovascular disease type two diabetes. It also puts the woman at risk for endometrial cancer. There's endometrial hyperplasia going on, which is why that's very interested in disease for my lab. And so the question is how do you study a complex endocrinopathy like PCOS there's so many factors involved there's different tissues involved. What is the ideology, I mean are there environmental factors involved. There are no animal models, you know, how do we do in vitro modeling. And so that was the challenge that we would put. And so this is our PCOS in addition lattice, where we have organ systems represented in our lattice including the ovary and tube in the endometrium as well as the liver, the pancreas, and we have space here to put other tissues as well. You know we always have to remind ourselves, it's conceptually it makes sense to put X, Y and Z tissues on this lattice but you have to build those tissues in vitro. And our approach is going 3D we do everything in 3D we have organoids, spheroids, X plants, tissues, etc. And so we feel that the 3D dimension mimics physiology better than 2D for what for certain things. So the first thing we wanted to do was to make a hyperandrogenic ovary, which is an ovary that makes a lot of androgens a lot of testosterone. And so how do we do that. When we look back in women and look at their hormone levels, especially the gonadotropes LH and FSH, we see that women with PCOS have elevated LH across the board, across the menstrual cycle, whereas normal in the normal menstrual cycle you see that peak, you see that increase and then a decrease of LH. And so we thought let's do that, let's add in high, high LH and high FSH right at the beginning. So that's what we did, we put in higher levels of gonadotropes to these mouse ovaries and then try to trigger ovulation. And what did we find? We found that these mouse ovaries were able to produce high levels of testosterone when we added these high levels of gonadotropes. And that was an amazing thing. And again, this shows how pliable, flexible the ovary is, how responsive it is to gonadotropes and how you can manipulate it to what it is you want it to do. So in this case, high testosterone. And what did this high testosterone do to the downstream tissues? Well, in the fallopian tube, one of the characteristic features is that cilia of the tissue, right? It's that cilia that waves the media along, almost like if you do have an embryo form there, then you want it to move along. But if you put in high androgens, here you can see PCOS like, what happens is that cilia stops beating, not completely, but the rate of beating of cilia decreases significantly. The other thing that we saw for endometrium, remember I said that women with PCOS are at risk for endometrial cancer. They do often have endometrial hyperplasia, which is proliferation over proliferation of those epithelial cells of the endometrium. And in vitro what we saw in our organoids of the endometrium that high testosterone did increase proliferation of the epithelial cells of the endometrium. And so we were able to mimic that in vitro. And then we still have experiments to do to see what does high testosterone do to the pancreatic islets or the liver in terms of their function and those are ongoing. The other question we want to ask is hyperinsulinemia of these women. Is it the hyperinsulinemia that are driving all of these dysfunctions even on the ovary? And so we have pancreatic islets that are able to release insulin, depending on how much PCOS that we challenged them with. And so we're going to start this experiment with these pancreatic islets as being the drivers. And so we can customize this lattice to say who's going to do what first and then who's going to influence the other tissues. And so this is a very versatile platform that we can ask a lot of these different questions. And so this is just a summary of what we are intending to do. We're going to take healthy as well as PCOS ovaries, which means high endogen releasing ovaries. We're going to add in healthy levels of insulin, unhealthy levels of insulin, etc. And see what happens to their function. And we can do this in the lattice system in a very controlled fashion to see who is influencing what the most. The other thing that I am really interested in is looking at how the PCOS ovary affects the endometrium. We can get benign endometrium, so not endometrial cancer, but benign endometrium from women that are undergoing hysterectomies, make organoids from them, and then expose them long term to high endogens. And we want to see, well, what happens to that endometrium, what are some of the epigenetics that are epigenic marks that are changing, etc. And so this is one study that we have where we're looking at obesity as a risk factor for endometrial cancer. And what we're doing is we're co-culturing fat steroids like in 3D with endometrial organoids over a period of 28 days or longer in the presence of hormones. So this is basically a pre-menopausal, obese women exposed to menstrual cycle level hormones. What happens to that endometrium or the endometrial organoid when this happens? And we can look at it at molecular level. We can look at potential mutations, epigen activation, epigenetic changes, etc. And so this is an example of how we can start studying chronic stressors when we are able to culture more than one tissue at a time for a long period of time. So this is a schema of what we can do with PCOS. We can also, along with the hormones of PCOS, add in environmental disruptors that are linked to PCOS, endocrine disrupting chemicals, etc. And so we are able to then, like I said, control the environment and control the factors that we're studying on these different tissues at the same time. This is from our collaborator where he can screen all these different endocrine disrupting compounds using just the ovary or the follicle cultures. And this is a very sensitive biological unit where, you know, it's amazing you think anything would disrupt the ovary or follicles, right? But it doesn't. There are only certain specific endocrine disrupting factors that affect the ovary. And then we can see then test some of these endocrine disrupting factors on the rest of the reproductive tract. And along these lines, yes, we can test drugs. This adds complexity to what's currently available in vitro. A lot of the drugs are screened pretty clinically using cell lines. But here we have 3D renditions of each tissue. And then not only that, we can put them together to make a more complex system to see how drugs are either metabolized or maybe retained, to see how that eventually affects reproductive tissues, etc. So there's a lot of possibilities that has opened up with the development of lattice. And so here I showed you how we can apply lattice to disease modeling. So as a summary, just being able to study a more systematic system. And understand how risk factors that are that we've known are associated through epidemiologic studies, we can now test them in the lab to see, yes, these particular risk factors such as that, or such as hyperinsulinemia. This is how it affects the downstream tissues biologically. We can test multiple drugs as well. And personalized medicine, I just glossed over it, but this is something that we're thinking of doing where we've started using induced pluripotent stem cells. We have been able to generate these stem cells from women with endometriosis to see if there is any genetic component of the disease that is contributing to the progesterone resistance that we find in these women, as well as the disease itself. So so much can be done. And so, building a reproductive tract in vitro. There's a lot of different choices that we can make in terms of the cell sources, whether we use primary or IPCC or cell lines or tissues, whether we organize men 2D or 3D or an excellence. And whether we culture them in normal 24 world plates, or in a dynamic tissue culture system, there are many to choose from, etc. But really the take home message here is, we have to allow biology to dictate what models we use we cannot force models on biology. And so each experiment is going to be context dependent. We need to know what we want to study what we think is going to happen, and then build models to answer those questions. And so that that's what I have for you today. I do want to acknowledge the team that first started in this project, Teresa Woodruff as the PI, and all of the, my colleagues at UIC and Northwestern, as well as Draper labs who really made that this event our system and set the ball rolling. And then our second part is the disease modeling tissue chip to part where we brought in people from Northwestern that are experts in PCOS like Margaret Urbanek reproductive immunology doctors such as Christina Boots, Teresa Woodruff's lab who started this. And then at UIC Joanna Brudette's lab that are experts in ovarian cancer and fallopian tube. And then Dr. Shau at Rutgers University, who does all of the compound screenings and from his follicle cultures. And so I also do want to acknowledge all the funding sources of NIEHS National Cancer Institute Office of Research of Women's Health and National Center for Advancing Translational Sciences. And so I guess that's all I have and I'm sure you guys have questions and look forward to having this discussion. Wonderful. Thank you so much, Dr. Kim. That was fascinating. So while I wait for some further questions to trickle in, let me just ask you just a few to serve off of for me. It looks like the lattice system will be capable of also modeling the male reproductive system a few the cell types is that true. It's not that we need any more research on males as you pointed out, but but it's possible right. Absolutely. And Teresa actually did start that and she called it the dude cube. She's really great at naming these systems. And basically, we're able to take testes tissues and look at you know testosterone function and production to study like azospermia, etc. And so yes, absolutely. If there are tissues available and a biological disease to study these, these systems are conducive for that. Yes. Okay, so I'm going to categorize some of the questions that have come in first I'm going to start with some of the more technical specific questions and then we'll zoom out to some of the broader social questions. In fact, Julie, if you want to open up the Q&A box, you can kind of follow on with this first question. It's a little bit technical from Ian, that's Sica. Yes, this is a technical question. Was the LH surge prior to P increase being artificially introduced or replicated or replicated in the mouse model with the larger dose injections given by the investigator. That's a great, that's a great question. Yes, the there are certain concentrations that have been figured out where we do add a lot more HCG a day prior. And after that what we see is a swelling of the follicle and actual the popping out of the O site. Can I share a movie that we just got hot off the press to show this. Yeah, go ahead. I am so excited about this because it's incredible. I'm going to stop sharing. Oh, I'm going to share here. So I want to show you this is time lapse over many days. And this is an ovary. As you can see the follicles are growing. Right. And then after we do you see that fall that O site coming out. Here's another one coming out. Here's another one coming out. It's incredible. I mean we have ovulation occurring in lattice. Isn't that amazing. This actually leads into a really excellent question is just coming from Jim bark Parker says even our lattice have been developed. Are they in general use by other labs. And if so what applications are being studied. Yes. And so we are using lattice in our lab. So even to our is available, but not currently being used for reasons that I mentioned how expensive it is and how difficult it is to use lattice is being developed and we do want to commercialize it we want to get it in the hands of the researchers. And so we use it in the lab. Other our collaborators use it, and you can do it's the system is tissue agnostic and disease agnostic as long as you have a question where two different or two plus different tissues are interacting in a specific disease, or it doesn't even have to be disease you can look at tissue longevity, long term cultures of whatever it is you're studying. And as I mentioned in my lab we're looking at obesity, and I mean endometrial organ noise to see what kind of changes occur in the presence of fat. We're also trying to figure out whether metformin is affecting the endometrial cancer cells or is it affecting blood vessels, or is it affecting the fat in the obese context. So those are really neat questions that we can ask and experiment. Yeah. So I have, I have a follow up question to that then. So it looks like the last system is going to be wonderful for interrogating all kinds of questions around drug safety drug screening. Can you modify the system in a way that you can look at drug safety for women who are pregnant. Right. So eventually, as long as the model systems are there we can, I mean, as you know from being in the tissue chip meetings there are placental biologists that have represented placenta and microphoetics and so placental function can be definitely studied, but in terms of crossing that placental barrier, do the drugs actually get to the fetus or not. Those are something that some things that can be studied. And what other things are involved in pregnancy, maybe in the early events of pregnancy, does it actually affect an implanting embryo. So it's a really neat study, not, not in microphoetics but using organoids from a group group in the UK, where they actually co-cultured embryos with endometrial, they call it assembloids but they're basically organoids to see how the that influences embryo, as well as different properties like expansion, for example. So these, these are being done in terms of the pregnant concept, but I think we just, we need a lot more work in that in building the models. But, of course, I think it is, it is possible. So let's move on to questions that relate to donors and sources of your cells. This is a question that Bridget Dooney is asking I believe on behalf of Caroline Lowenthal, hi Caroline. Some women know they don't want to have children or know that they're done having children. Are these women ever considered as a source for biological samples that could otherwise compromise fertility. We don't. We just get access to tissues from the OR from women undergoing surgery for whatever indications. I'm not sure what the channels would be for obtaining tissues like that. I'm not aware, maybe other people are, but I guess once if a patient wanted to donate their tissues, that would be something. But we, we, we don't do that. No. So Julie, I have a question for you. All the cell types that you have on the lattice, do they have to come from the same donor? No, no, right now what we have are different cell sources, all human except for the ovary. And it is, it is a challenge when you're not working with a hospital and the number of surgeries are not there, especially during the pandemic. It was, it was very hard to get any kinds of tissues, because these are electric surgeries that we get a lot of tissues from. So, no, to answer your question, no, they don't have to come from the same person. But there is something to be said about personalized medicine. Like I said, it'd be great to see all of these different tissues being made from one patient source, and to see how that particular patient responds to whatever is hormones, drugs compounds, etc. Right. So I'm thinking that. So I was really fascinated by what you said earlier about sort of like women's participation and enrollment and research in the past and the lack of proper adequate involvement and numbers of research subjects that represent female biology. I think there may be actually through the lattice system, even like a greater opportunity to expand the scope of who gets to participate in this kind of research to populations that are typically underserved, for which it would actually even be difficult to put them as human subjects in the trial people are like far away places or people who are resource poor settings. If you just need their cells, if you just need, you know, cells that represent the biology to them put into the last system that could provide a possible way to enroll a more broad diverse range of research participants, and kind of doing you know, in-person clinical studies in a particular cohort. I mean, or do you think that there's an opportunity to use lattice, like the lattice system to sort of expand, not just, you know, greater involvement of women whose biology is represented in studies, but just among women, different types of women who typically are not enrolled in studies or don't get the opportunity to participate in the study because you're using this. Yeah, I mean, I think the possibilities are endless and I can totally see in the future after we are able to make all of these different model systems to do just that. Like I said, lattice is just the plate that allows communication between wells. Like I said, yes, we can simply get blood from women and then make them into IPSEs and then make different organ systems and then put them on lattice and study them. We're also trying to figure out if lattice is conducive to differentiating the IPSEs into the different tissue types, only because I don't know if anybody has worked in that field, it's extremely labor intensive. They require handling every single day, no holidays, no exceptions, and some of them require hundreds of days before you actually get the final tissues. But if we can use lattice to kind of provide that daily nutrients and stimulants without somebody being there every day, maybe give them a two or three day weekend. So that's something that we're also working on as well, which is really exciting. So a lot of these things are very possible. And I think the fact that you did identify those needs, I mean, I don't even think about things like that. I mean, that is such a great need to involve patients that normally wouldn't come into the clinical trials. Just get some blood from them and just do it. I mean, that is so cool. Yeah, I mean, I was thinking there are so many barriers to participation for people who are underprivileged. I mean, they can't take time off sometimes to even be in the study. So if you just need healthy tissue from them, and it's not even like they have to give up a major part of their body, they can just provide cells, somatic cells. That could really be quite revolutionary to get the kind of data that's rich for a population that's kind of got that genetic diversity you're hoping for. There are some really interesting possibilities. Let me ask you further of the lattice system. So it sounds like it's much automated, like there's that doesn't have to be somebody there 24 seven overseeing. It's not correct. It's sort of the you can sort of let it run on its own. Correct. I mean, you always we have a system where there is there's a well that has media and you have about three mils of media and that depending on how much media you want it to flow through has to be replenished. And that can happen over a course of two or three days. So yes, it can run for for a couple of days without you watching it. Right. Well, yeah, I mean that the time of labor intensive nature of, for example, I PSL research. When you said that it just ran right and he doesn't want to be involved in that because it's just like you need said not being allowed all weekend long. So it's very it's very labor intensive but if you could do a lot of this work through an automated that could really, I think make changes to the workforce and sort of Yes, involved at the lab. So you don't get people like my son saying I don't want to go to that field because I can't I don't want to go in every day. It takes a certain person to dedicate every day to come into the lab to babysit this. Yes. Yeah, yeah, absolutely. So, I'm wondering. So let's get on to this workforce issue. There are a lot of young kids who are interested in STEM, who say they want to be a doctor or they want to be an engineer. I don't think a lot of them not yet anyway think that they want to become somebody who works in developing micro physiological systems right like how do you how do you sort of get the word out like, how does one find out about this besides going into initially biology cell biology and then learning in graduate school the kind of work that you guys are doing. How does how would somebody learn to get this field on the radar with a with a kind of like hear about this through engineering. How does a young person like find out about what you do. I really think young people know so much more about technology and where to find things and the avenues that they have are just incredible. I think if we do a good job on just highlighting what what tools we have into to use in the lab. That'll disseminate through like social media, Twitter, etc. I think, you know, I think as light as the word gets out. It's, it's so fascinating. And I think somebody would be immediately drawn to something that can be to do so many different things for the betterment of physiology and for diseases etc. I don't think it would be too hard to to really represent that in just overall. Where would these young kids hear it from. You know, other than what I just said, I we we also host you know, high schools to come in and take a look for a day, what we do. And there are many programs that we can do for that too. I don't know what I don't know. I have this question on my mind a lot because I'm doing work in the Cambridge Boston area with a lot of companies that are struggling to sort of develop the kind of workforce that they need in the future for these kinds of technologies and a lot of people don't really realize that this is like an open new area of research using micro micro floating systems, exceed the modeling using cell types from donors. It's not in the traditional kind of radar of people thinking about medicine and research and human subjects research. It's got it's quasi engineering is quasi cell biology is quasi medicine is kind of got a lot of that. Can you explain a little bit describe a little bit more to me your team members like when are their disciplines. They're obviously collaborated with lots of people who, who's typically comes together on a project like this and what are their backgrounds, how did they get together. Yes, my lab is primarily made of cellular molecular biologists, but we do have bio engineer postdoc that we recruited. We did work with another bio engineering student that was in the bio engineering field at Northwestern. So it's very important to collaborate with those bio engineers. We did partner up with engineers at Draper. And it was a very interesting interaction. I mean we learned so much from them and vice versa. It's a different language, but I feel like those, those borders or those boundaries are being blurred more as people become more interested and they extend out. I don't think you have to have a bio engineering degree to be able to work on micro fluidics, and etc the you don't have to be a strict molecular biologist to understand what bio engineer can do with their systems, as long as them, the team interact and closely together there's so much information out there as well that we can plug into so I do have a diverse team. And I do, I do work with experts in each of the areas to make sure we're doing it right. I was also fascinated when you were describing the development of these technologies that what you have to use our materials that don't bind with hormones. How was that discovered was that through trial and error. I thought that was really fascinating. Yeah, I believe so, but the PDMS, I believe we had tried it very early on and we were, we were not getting the hormone levels that we should have and then you know we found that yes, those hydrophilic molecules to bind to PDMS. And so it was that was a challenge of looking for a good material to be able to to make these fluidics. And so we just went back to to what everybody uses in cell culture is polystyrene. And that's inert. And so that's that's what we based our construction out of. But it is, it is an area like a lot of the micro fluidics out there the MPS is are made out of PDMS. And so there are limitations exactly as long as you're measuring what the tissue is seeing if it's sufficient I think those are some of the precautions that need to be taken. And we do need to be aware that some of these materials are not good and finding that some of these materials actually leach toxins. I mean if you're working with cells that are pretty robust and you know they're there are hardy. They don't really show too many signs of these toxins but then you're working with brilliant sensitive cultures like the ovary or the follicles. And yeah it matters that matters what materials you use. Yeah, yeah I have a few more technical questions and then I'll get to some of that the broader ethical issues in a minute. So in the very early days of tissue culture research people thought that light had an effect on the tissue culture so people were like dark clothes maybe like in darkened rooms. But are there, this may be a hard question for your answer but are there like concerns you have about other ways in which you may be creating artifacts in your system that you're constantly on guard for how do you, how do you scan the horizon for those kinds of concerns So I suppose that the system is kind of a body temperature and it's kind of got like it mimics as much as possible the human body. Are there other like anything else besides the materials that you're using and whether they're leaching toxins or finding with with hormones or the other things about the system that you're kind of worried in the back of your mind, maybe this is doing something we don't want it to do. Yeah, yeah, of course I mean that that that is always at the back of our minds, other than the actual materials or the environment okay so there's one thing in the environment and that's not just our micro fluidic environment but just cell culture in general. So we do put them in 5% CO2 right and in the body there's a lot more than 5% CO2 and that that's always been kind of strange to me but that that is the standard that we use for all incubators. And that's that's what we've done. You know what the things that not concern me but that are at the forefront of my mind is the missing cell types right in the body you have immune cells that are really important they do so much more than just to protect you from infections. You have biological roles in each of the tissues, especially if you're talking about the menstrual cycle and the tissues, the changes that are occur in response to hormones, those immune cells are really important in contributing to some of those changes right blood vessels we don't have any blood vessels in our system and they're really important, especially if you want to see bleeding, if you want to see all the blood cells, you know, degrading at men's sees, you know, so, so there are certain cell types that are missing. We don't know how to incorporate them yet into our system, but we think that they will be essential to really give us a more complete view of what what's happening. I guess I apologize for my audience have a few more technical questions and then you know, we'll jump into the other side. So, so the other question, are there any mechanical, or other like pressure aspects of the system that you need to be mindful of so the microfluidic system obviously you can control things like pressure and mechanical forces is there is there much thinking that has to go into that. So, that's we really wanted to make a more simpler system so it's the computer that really directs and controls how much flow goes by how much. This is a question for my bioengineering post actually, but while we were building lattice we did run into a lot of those problems to figure out what is the good flow rate that is representative. The pressure, the hydrostatic you know differences, depending on your your culture system and the design. What happens when there's extra media that's left over and you know you're rotating it like for for days, is there is there going to be a reduction that needs to occur. You know these are all things that definitely need to be worked out when building a system like lattice and I think we've run into every problem that is possible and finally we're at a working place right now, and it's working really well So, did I answer your question. Oh yeah, you did. So it is, it is, it is true that there's a complicating factors that yes, yes. Are you, are you submitting to FDA or have you submitted to FDA this lattice and, and if so what application are you, if you can, if you can speak about that. So submitting to the FDA just just like for FDA approved, this is something that needs FDA approval is it going to be a diagnostic like how do you sort of frame this to FDA. I first want researchers to use them at the bench. We're going to collect more data we did that's what we need more data, not just in the reproductive system but data like anywhere to see how useful it can be. So we want to make more of it and train more people on it. My postdoc always says I can train you in an hour to use it so it's a pretty easy system. But even before we get to the FDA but we eventually do want it to be able to be used as a dark screening tool, in which case then we would have to go through those regulations but I feel like we kind of need to gather the data first. Yeah, I suppose that you would have to confront this question, or you are confronting this question before you even get to the FDA if you're going to have other researchers use the system. But the question is, how confident are you that this is actually recapitulating the human biological system. Like how do you actually validate what lattice and event are doing against clinical the real the real deal the real human body is what are your reference points know that this is actually doing what you're hoping it's doing. Yeah, that's a great question. I was saying that this system is totally recapitulating what you see in the body, because when we are missing some of those key factors and, you know what we're pushing through is media, and media is not blood. We would love to put in blood sometimes you know to try to figure out how we do that. But those are some of the things that we need to tackle first. Could you, could you just repeat your question. Yeah, so I mean so what are your benchmarks for for. Yes, yes, whether it's physiological or not. Like I said, we do know a lot that goes on in vivo, because we've been able to get at least samples from women, at least you know in different cycles, but also from those animal studies, we can, we can, we know what needs to happen. So we take all of that information as our frame of reference, all the in vivo studies, and then we compare it to what we have done in vitro from what we've had. And we do see differences that recapitulate more the in vivo than the in vitro. And so we're, we find that we're kind of moving in the right direction, at least. At least it's it's a bit more physiologic we're not saying it's perfect. There's a lot more work to be done but we are getting new discoveries and new information because these these are in more natural environment as well as a more natural architecture and shape. And then last technical questions come in, anonymously, because sharing media between the organ types helps them grow or become responsive. How do you think common media changes at once every three days are influencing the system. Have you tried refreshing only half the media to see if it yields any benefits. Yeah, they're talking about conventional cell culture media when you change media every three days. Instead of culture. Yeah, definitely, depending on your cells and how metabolically active they are. They're going to be sitting in waste products and stresses. They're not going to be seeing the additional nutrients, the fresh nutrients because they've used them all up etc. So I think there are there are changes or responses that you're going to see because of cell culture artifacts. We have done some studies to show that comparing static and micro fluidic, where the, the availability of glucose is different to the, the health is different of the different cells and tissues. And so definitely there there is a difference between static and micro cultures. So if you do any ask, what were, what were you most surprised about the results of your study, or these types of studies. You know, when you do a study like this and one of the reasons why it was so fun was because it was not really hypothesis driven but it was basically building something and saying let's see what happens without much expectation. What I personally was very surprised about was how all of these different tissues from different human sources, as well as the mouse ovary, were okay in one media, and not only were they okay. They responded to those hormones a lot more robustly. So I think, going into this, we knew so little about all the parameters of each of our tissues and isolation that are so important for the other tissues. And so, I guess it was more naive on our part, but that was what surprised us the most health physiologic I put those in quotations, more physiologic these tissues were behaving. So let's see and hear about the evolution from the guitar to lattice what's what's sort of the next generation what's coming after lattice. Oh, Wow. You know, if there was a way where we can be more high throughput. As you can see lattice had eight, eight different compartments. And if you want to start testing drugs, you would need many more samples they're represented there. I'm not sure how I would do that, but that would be again, I guess the next need for the field is to be able if we're confident enough to be able to test multiple drugs and multiple concentrations and then comparing that. So I'm not saying that these MPS systems are going to replace standard 2D. I think we need all of these systems to give us to inform us of how the drugs are working, depending on the context experimental design right. And so I'm the believer that you know the more information that we get, the better. So is it going to eliminate mice studies, I don't think so it might reduce the number of mice that are used in research but I think those in vivo studies will inform us. Again, whether differences in the drug happen, different responses happen in the microfluidics versus the mice and if so, why you know, so I think those are all important questions as well. So you mentioned that that you're, you are now sharing the lattice technology with other labs is it was really also very interested in how you were trying as you're developing this technology to keep it cost effective to keep it kind of extra friendly for other other labs. Is there a little bit more thought given to trying to make this a useful tool for people in lower resource countries labs that are kind of less resource in Northwestern people in other other countries who might want to do this kind of work. Is this technology going to enable them to do a higher level of research in their more limited research setting at their institutions. Was there a thought and that kind of like you know democratization of the technology. And again you bring up such interesting questions and I think yes, the question is we would love to see that. You know we did. And that was our goal to make it to make anybody be able to afford something that they want it and that's why we went with 3D printing, right. But unfortunately, that didn't work out but somebody can find a material that is is great for 3D printing to print out these systems that would be even better. But yes, that was our goal to make it user friendly for not just Northwestern people, but for everybody across the board. And we are sharing it with just our immediate circle because we don't have enough units right now. But the goal is to partner up with a company that enable us to make more units so that the cost can go down even more potentially and then share it and the training again. It's very easy is just, you know, telling what the scripts to write on the computers and, and it'll just run so that that's our ultimate goal. So one last question. Do you have any ethical concerns about this technology anything that that you you think we should keep our minds on. Again, this is just a technology is just a tool plate. It's it's something that you can control. I think the ethical questions come in when you're asking about the experimental question. What is your experimental question. Are you going to be needing tissues that normally you can't get or shouldn't get. I think the ethical questions are there. And I think that's where you're the expert on these. But again, this is just a tool is like a cell culture plate. A second generation that allows you to do a lot more things than you can conventionally. And that was my way of skirting the question that I don't really know. That's wonderful. I mean, it's just the technology is why I invited you for this series. Thank you so much for your time. Thank you so much for leading us through these wonderful developments. Good luck with your research going forward. So I'd like to conclude our session for today. I want to thank the Center for bioethics at Harvard Medical School for sponsoring this event. I want to thank actually Troutman and how SF and I just for all the logistical support in this series. And I want to thank you our audience for joining us today for this session is hope you've learned a lot I know I did. Please join us next fall when the session when the series continues, and we'll have plenty more to talk about next fall. So until then, thank you for joining us have a great weekend. We'll see you next time. Thank you so much.