The Liver Meeting 2022-Hans Popper State-of-the-Art Lecture

Hans Popper State-of-the-Art Lecture

Video Transcription

Good morning, dear members, guests, and colleagues. I'm Jorge Bezeja from UT Southwestern and Children's Health in Dallas.  It is my distinct pleasure to introduce the distinguished speaker who will give the 2022 Hans Popper Lecture. Dr. Popper was a legendary hepatologist, pathologist, and teacher.  He is one of the founding fathers of our specialty, hepatology.  He was born in Austria, and in the Second World War, he immigrated to the United States and started his life in Chicago.  At the University of Chicago, he first focused on, can you put my first slide, please? He first focused on obtaining a PhD focusing on pathology research.  After joining practice, he evolved to lead several, to chair several leadership positions,  including the scientific director of the Houghton Institute for Medical Research in Chicago, where he held the first hepatology meeting in 1948 with a group of colleagues.  And two years later, they founded an association that is today known as the American Association for the Studies of Liver Diseases.  Unfortunately, you don't have the slide that I prepared with Dr. Takebi,  but it is in this context of scientific and academic excellence that I have the distinct privilege to introduce you to Dr. Takenori Takebi.  Today, he is Associate Professor of Pediatrics and the Endowed Chair of Organoid Medicine at Cincinnati Children's Hospital.  He is also the full professor at Tokyo Medical and Dental University in Japan. Dr. Takebi obtained his MD from Yokohama City University in Japan,  where he became Associate Professor in 2013, shortly after his graduation from medical school. He joined Cincinnati Children's three years later in 2016.  During medical school, he carried out pioneering studies using iPSCs to engineer a liver organoid to investigate development and to model human diseases.  He is one of the founding leaders of the Center for Stem Cell and Organoid Medicine at Cincinnati Children's.  In addition to holding several grants from industry, from the New York Academy of Sciences and Foundations, he currently holds a prestigious NIH Director's Innovator Award.  His work has been published in journals of broad readership, including Cell, Nature, Nature Medicine, Cell, Stem Cell, and others.  Dr. Takebi has received several national and international awards. Perhaps more telling is that he has been the subject of interviews in the journal Science,  and the title of that interview was Mini Livers Reveal the Fine Details of Organ Development. And in Nature Medicine, the title was Creativity for a Cure.  His scientific contributions go beyond hepatology.  For example, during the peak of COVID-19 pandemic, he sought to develop other ways to save lives of patients with respiratory failure.  So, beginning with rodents and then transitioning to pig models,  he produced comprehensive data in proof-of-principle experiments showing that the intestines can be used to oxygenate animals in respiratory failure.  This study was published in MED and featured in the New York Times and other late media.  Today, he will be himself, I hope, as a hepatologist discussing how human liver organoids can be used for drug development.  Members and colleagues, please welcome Dr. Takebi to give his talk. Thank you.  Sometimes this happens when we go from all-virtual to in-person. Our apologies.  So again, my apologies on behalf of the association, Dr. Takebe. All right. Wow. It's truly a privilege for me to just to give, you know, such an amazing opportunity.  And first and foremost, and thank you so much, George, you know, with the wonderful introduction without the slides.  And also, you know, President Loli DeLev and also the leadership members and all of the participants for giving me this fantastic opportunity to share our latest work related to human stem cell organoid based approach towards a future hepatology advancements.  So my name is Taka Takebe from Cincinnati Children's Hospital and also University of Cincinnati and where Dr. Leon Schiff was actually studied in our university.  And I'm very honored in honor of Dr. Hans Popper to address my, you know, named lecturers.  But I was actually studying a lot about history of Dr. Hans Popper, but already Georgie has beautifully summarized his history.  So I probably don't need to spend time on explaining Dr. Hans Popper. He's a founder and father of hepatologies. So today.  So this is my disclosure, but I'm going to skip for the sake of time.  And today I'm going to, you know, introducing the concept of the human organoid based approach with a focus on this particular organs of interest, as many of you do as well.  So this is a liver, as you can appreciate from this beautiful live imaging analysis.  There are beautifully organized a variety of different blood vessels and perfusing lots of, you know, immune cells, metabolites and lots of key fractures to trigger the most homeostatic renewal of the liver tissues.  And not just that, as many have heard a beautiful talk from many speakers at the ASLT, there are a variety of different hepatocytes ranging from zone one down to zone three.  And even more diversity is recently appreciated by the single cell RNA sequencing approach.  So the way we are approaching this is try to make a functional human liver liperator in a petri dish so that we can study or trace the process of development as well as disease progressions.  So that's our overall questions we want to test. So the way we are approaching this is to utilize human organoid model system.  So organoid can be defined as the 3D tissue structures or organized tissue structures that contains a variety of different organ specific cell types, but not just presents.  But we need to have a structurally organized components, for example, like cellular polarities, epico-basal polarities, and as a, you know, basement membrane production.  So that sort of, you know, spatially organized feature is also one defining feature for the organoid.  And because of this, we now are able to monitor the organ specific functionality.  For instance, in the kidney case, maybe uline production filtration processes can be detected in the organoid culture.  Same for liver, lots of sealant proteins is processed towards the culture media, whereas bio-acid is eliminated towards the bio-canonical side in organoid model system.  So we can play around a lot about the hepatic functionalities as well as pathogenesis process.  So just to summarize how organoid is evolved and where we are now in the space of organoids.  So organoid can be either generated from the primary patient samples or adults than progenitor cell populations.  So if you can get a biopsy of the patient samples from like intestines or liver, you can nicely glow the patient specific either healthy or diseased organoid model.  That has been tremendously led by the Dr. Hans Krieber's group at the Netherland.  And nowadays, lots of other, you know, pioneering faculties doing this adults than progenitor-based organoid generations.  The other approach which we are primarily working on is a human prepotent stem cell-based approach,  wherein we generate iPS cells or embryonic stem cells from the humans to directly differentiate it into the variety of different organ system in our body.  And this work is particularly pioneered by Dr. Yoshiki Sasai in the field of Blaine's or eye organoid formation process.  Now followed by lots of, you know, key leaders in this group including Dr. Jim Wells of Cincinnati Children's Hospital and Australia for the kidney,  Marissa Little for the kidney organoid generation to name a few.  But nowadays, almost, you know, entire all of the organ system can be glown as organoid system ranging from brain down to the bladder or prostate components.  And either from adults themselves populations or prepotent stem cells.  And more importantly to the hepatology communities, these are, you know, preserving some of the patient's phenotype from a diseased condition.  So we can potentially make cancer organoids as well as disease model system by, you know, triggering the pathology stimuli in a culture conditions.  So one of the major differences you need to appreciate between these adult stem versus prepotent stem cell-based approach is diversity.  So adult stem cells is really, you know, recapitulating the patient's conditions, but unfortunately they are creating predominantly for epithelial cell types.  Whereas the prepotent stem cell-based approach has the capacity to diversify into the different lineages including the mesenchyme populations, immune cell populations, and as a population as well.  So using the prepotent stem cell-based approach, maybe we can test a variety of different tissue or cell type contribution in our culture.  But regardless of these differences, we can now generate human stem cells from adult populations as well as prepotent stem cells that preserve the patient information  so that we can test the human-specific mechanistic angle in the pathology.  And in future, potentially we can personalize medical applications or helping the industry to identify the better drugs to facilitate the drug development process.  And in the longer term, we can transplant the healthy organoids for the patients with endostage liver conditions. So that is the sort of future dream of our organoid-based approach.  I hope that at the end of my talk, you might be excited about either of the potential applications of this organoid medicine potential.  So we are primarily, as I said, we are primarily working on the human prepotent stem cells.  So the way we are creating the human organoid model is always facilitated by the, you know, decades of studies around the development of biology.  For instance, early, you know, fertilized egg is actually divided into the multi-germ layer, including like endoderm, mesoderm, and ectoderm.  And this endoderm is actually giving rise to most of the digestive tracts.  And that process is pretty well studied in a number of key animal model studies, including like fish models or frog models or mouse model or other models as well.  So these decades of important studies is really hinting us to, you know, study the soluble factors or cellular factors which is driving the each step of organogenesis process.  For instance, my close collaborator, Jim Wells and Alan Zorn at Cincinnati Children's Hospital,  spending almost 20 years to understand how liver-pancreatic digestive tract is formed from the prepotent stem cell stage.  So we now are in an exciting time to really manipulate these pathways to direct the human liver formation process.  For instance, liver is actually emerged at the boundary between the foregut cells as well as meat gut cells.  So we think that this sort of positional cue is very important to pattern or generate the liver biliary system.  So to test this idea, really talented Pastor Ken and Hilo was really spearheading these challenges.  So what they have found out is to specify the foregut cells in blue in these movies as well as meat gut cells with the green in these movies.  After making a boundary, providing a spatial cue that is found in a normal development,  they have discovered hepato-biliary pancreatic progenitor is actually specified in the border of these two tissues.  And after figuring out how to culture them in the longer term, applying the air-liquid interface culture,  they have found out the process of imagination, process of budding, process of the hepato-biliary domain segregating to the liver,  as well as extra-hepatic biliary system, as well as pancreatic system. So not just, you know, the presence of each organ domains,  they have also confirmed after exposing the fluorescent biocid in this culture conditions, they could figure out the connection in each other.  For instance, in this particular experiment, they have exposed a CLF that is uptaken by the hepatocytes at the red color in these cartoons.  That is eliminated towards the biliary regions as well as at the end into the duodenal structures. It's a little bit of exaggeration, though.  But also, they have tested the genetic fidelity to the animal model by targeting S1, which is a notch effector genes that if disrupted, they are known to cause biliary abnormalities  that is converted to the pancreatic tissues. And that can be also confirmed in this model system. So at least with these experiments, we could confirm this in vitro model system  is actually recapitulating what's seen in our animal body. So overall, there are a number of critical protocols just to navigate into the blood  or different liver components to be modeled. So I think I would call it as a modular differentiation, because I think if you want to push, for example, kinetic life formations,  we need to expose linoic acid to induce the biokinetic pathways. But if you persist this, maybe you're going to lose the hepatic identity.  So there are lots of pros and cons depending on the protocol. So what you want to see is really defining the protocols to generate, for instance,  like extra hepatic biliary systems or maybe hematopoietic progenitors, alkyl cultures. And more recently, we figured out the sympathetic nerve can be also induced  in our organ culture system that modulates the gluconeogenesis process by sympathetic nerve. But what I am going to try to address today is more of the complexities,  including organ-specific vasculature components. So as many of you know that sinusoidal endothelial cells is really key for a number of important liver functions,  but never introduced in this type of organ and motor system. So Dr. Saiki, a really talented postdoc, is really tackling these problems. So we previously, in 2013 and 2017,  showing the three different progenitor co-culture systems is actually enabling the vascularization of the liver organoids,  including hepatic endoderm, mesoderm, and arterial progenitor populations. But what we are missing in that publication is the sinusoid.  So Saiki actually looking back the normal development by looking at the single-cell transcriptomic signatures of developing human fetal vascular networks,  and finally finding out vitelline vein progenitor cells, which is a connecting vessel between the yolk sac towards the sinus spinosus,  is pretty much precursor for the sinusoidal vessels in normal development. And also they are Wnt2-expressing populations.  So what he has applied is, in addition to these three ingredients, he also layered, overlaid these sinusoidal progenitor cells  at the middle of liver organoid formation to see what's going on. And what we have discovered is quite interesting. For example, this is a time-lapse imaging  to just show you the vascularization or network formation from the endothelial population. But going to the down pictures,  you can nicely appreciate the heterogeneity of the blood vessels in this culture. For instance, when you look at the CD31, this is a panendothelial markers,  and entirely expressed in the network of this entire organoids. Whereas when you look at the CD32B, which is more towards the sinusoidal-specific markers,  that is only expressed in the middle part of this organoid culture system, which is probably around 5 mm in size. When you look at the surface by electron microscope analysis,  you can nicely appreciate the fenestrated structure that is completely missing in arterial progenitor populations. We also confirmed some of the functionality as well.  But what we are so fascinated after this sinusoidal integration is in the next slides. For instance, when you do the sectioning analysis of the HE in these 5 mm tissues,  you can nicely see the hepatocytic population is inside in this organoid culture, which is very gigantic. But when you look at the zonal-specific markers like GS and GLS2,  you can nicely see the segregation of each population. For instance, in the lower pictures, you can see the live one positive sinusoid is distributed across the entire tissues.  But GS positive pericentral population is actually located adjacent to the location where we put the progenitors on the bottom.  So there are nice segregation of zone 1 and zone 3 phenotype after sinusoidal co-culture-based approach. And we're also looking at the functional outputs,  including like CYP cytochrome activities as well as glutamate synthase activities that is specialized for the pericentral population. In the previous arterial endothelial culture,  it does not provide much CYP activities as well as GS activities, but they can be highly upregulated after sinusoidal co-culture.  And the mechanisms of that is likely due to the Wnt2-based crosstalk activating the pericentral activities. For instance, when you target Wnt2 in a sinusoidal population,  you're going to quickly lose the GS positive hepatocytic population, indicating this endothelial population is actually helping to create this donor specification profile.  But perhaps most important sinusoidal function is, I believe, is the coagulation factor synthesis. For example, factor VIII is actually produced by the sinusoidal endothelial cells.  So to really test the functionalities of the factor VIII synthesis from the sinusoidal vessels, we created the factor VIII deficient knockout  immunodeficiency animals and tested the transplant efficacy. On the panel B, a suggested confirmed human CD32V, which is a human sinusoid, is expressing factor VIII  by blue color, whereas the mouse CD31 positive cells is completely missing the factor VIII as confirmed by immunostaining. And panel C and D is actually looking  at the breeding phenotype. It's completely reversed in this organotransplanted mouse, as we've seen in a normal therapeutic approach, which  is the recombinant factor VIII infusion methods, whereas in a sham treated one is continued breeding from the tail cut vein. And panel D is looking at the APTT.  So the beauty of this system is you don't repeat the injection of the factor VIII. So even at the five months after transplantations,  you're going to see massive effective prevention of breeding phenotype as seen in repeated injection of recombinant factor VIII synthesis protocols.  So we think that this sinusoidal vessels is capable of performing some of the key sinusoidal functions. I just wanted to introduce one unpublished and uncompleted  work, because there are lots of implications. So this one is actually related to immunosuppressant-based drug-induced liver toxicity. So one hepatologist really applauded me  about the patients who manifested with the anti-cymoglobin infusion after transplantation to suppress immunorejections. But it turns out the patient was severely  induced for the drug-induced liver damage, with elevated STLT and elevated bilirubin. So he really wanted to try to understand what's the pathologies behind this ATG-mediated liver  toxicities. So we can leverage our sinusoidal liver organoid model system to test that out after transplantation. For instance, this on the left panel  is just to show you the engrafted liver organoid with sinusoidal populations.  And when you expose anti-cymoglobins, you can nicely see the breeding and coagulation thrombotic phenotype in the transplanted locations. But not just that macromorphologies,  you can nicely confirm lots of immune interaction, including neutral field visualization by light 6G antibody, and also platelet interactions by CD41 antibodies.  So you can nicely label human LCEC population by green color with the blood perfusion by blue color. You can dissect the neutral field platelet interactions  to trigger this type of sinus obstruction syndrome conditions. And that is really specific to human blood vessels. And also ATG is specifically uptaken  by the sinusoidal population, found in this and other live imaging analysis. So I think that combination of organoid transplantation with the intraviral imaging approach  might be effective ways to look at the immune interactions to trigger some of the pathogenesis. So I just wanted to highlight this as an example of the advanced disease model.  So now I'm going to switching over to the NASH. And Dr. Mora is already beautifully summarized. So I don't need to make a quick deep introduction for these disorders.  But one thing I want to appreciate is all of the patients is really related to insulin resistance, as many of you have already learned from a number of beautiful talks.  And also, we need to appreciate, for example, in the Asian population, relatively non-high BMI population also showing the insulin resistance with the NASH phenotype.  So we clearly need a more personalized basis to understand what's the differences between these type of different pathologies.  So I think organoid can fill the gap to some extent, right? So animal model is really crucial for the more systemic crosstalk. But it's a little hard to actually see the hepatocytes  in animals or even in humans. And this is a problem because, as Dr. Mora kind of summarized, adipose tissues or muscle tissues always helping to de-steatosis of the liver,  and also environmental perturbations and comorbidities, those ways confounding the interpretation of the imbalance in energy intake. But in organoid culture, you can specifically  fix the perturbations in terms of genetic level or non-genetic level. So you can test the mechanistic consequences on the metabolic dysfunction.  So that's why we think organoid-based approach can be a complementary model for the human NASH studies. So to tackle this problem, we need  to solve the problem of the model complexity. So Lien actually spearheaded this problem by developing the protocols where we can generate  not just hepatocytes, but also macrophage-like population, like copper-like population, as well as fibroblastic stellate-like population in culture after 20 days from IPO cells.  And she went on to expose very fatty culture or hyperinsulinemic conditions in the culture media supplements to induce the progressive NASH-like pathology.  For instance, when you look at this live imaging, this is like a live pathology analysis, looking at the highly-statotic organoid, or SHLO,  versus normal HLO, showing you the massive body PY, which is triglyceride equivalent uptakes in the liver cells. And then also, you can test the ballooning-like phenotype,  including the cytoskeletal filaments deposition in a culture system. So you can nicely see the pathology consequences in a live manner. And when you look at the day one tissues,  you can see those dependent uptake of the lipid accumulations. And on day three, because of macrophage contributions, we could see the number of key inflammatory cytokine  productions. And down below, at the day seven, you can see the fibrosis deposition in this culture system. So essentially, we are shortening or accelerating  the process of NASH fibrosis transition process within seven days. So we also like to note that this model is probably not reflecting entire NASH populations,  but rather specific to metabolic dysfunction associated NASH. Because when you look at the insulin response, like euglycemic clamp analysis, we  can do the same thing in a hyperinsulinemic culture condition of the organoids. When you look at the HLO normal organoids exposing insulin, you can nicely see the separation  of the glucose production. Whereas if you induce a SHLO condition, a set of hepatitis conditions, you're going to lose the insulin-dependent glucose  production suppressive capacity, indicating this is insulin-resistant. And also, a variety of other energy metabolism pathway, like fatty acid oxidation process is impaired.  And also, looking at the oxygen consumption rate, indicating mitochondria is somehow disturbed after SHLO production, as we see many in patients with type 2 diabetic NASH.  So in essence, SHLO in this particular culture condition likely reflecting NASH plus type 2 diabetic condition in the patient's conditions.  So we next went on to study the differences in the patients.  But the question was, I think this type of model is too complicated to compare patient-to-patient basis and not scalable. So we want to scale it up to the level  where we can test the population-level analysis. So Masaki has very creative ideas to really scale up this analysis. So he has generated 24 different IPU cells  in making progenitor cells and mixing them exactly in the same well in the same drop-off of the materials and exposing fatty acid and hyperinsulinemic conditions  to determine its identical conditions, how many diversities in each donor presents. So what he has looked at is initially on the steatosis by looking at the live imaging analysis,  then correlating back to the SNPLA analysis data  to perform organoid G-ROS type of analysis. What he has found out within just a tiny sample size of 24, he could find out the PNPLA3 is positively predicted  as a risk factor for the NASH in this organoid model system. But also he could find out that GCKR is also the major driver for increasing the steatotic conditions.  So we next try to understand how this genetic variance is influencing. And I just wanted to introduce Hans Popper here again, because he was the big fan of tackling  the controversial question in the hepatologies. So based on the learning from Dr. Hans Popper, I decided not to pursue PNPLA3, which is already kind of proven  and very positive for the NASH prediction, but we rather prefer to look at the GCKR because this is very controversial. So just as a background, GCKR, Glucokinase Regulator,  is actually a known factor to promote NASH, but also protective for type 2 diabetes because of these mechanisms. So GCKR is actually preventing the glucokinase activation,  but when you have a risk allele noted as TT, GCK, glucokinase, is highly abbreviated in the cytosolic locations, meaning glucose is actually utilized at the top,  whereas at the downstream, this was used for de novo lipogenesis process. So it's kind of interesting, right? Because this is protecting type 2 diabetes,  but promoting NAFLD, which is often complicated each other. And also this variance is extremely common across the population, except African American populations.  So most of the population is representing like 40 to 50% distribution over the risk allele carriers.  So we decided to look initially on the mechanistic level, looking at the glucokinase activities. So when you look at the GCKR TT patients  or plyme edited or gene edited CC to TT edited ones, you can nicely appreciate the glucokinase activity increase relative to CC, normal different cell level.  So this is okay, confirming this scheme. And in addition to that, when you look at the lipogenesis phenotype or stethosis phenotype, we nicely appreciate the differences,  clear differences between the normal or allele versus gene edited ones, as well as de novo carrier of the TT variants, suggesting lipid uptakes or lipid production is enhanced.  But to really confirm this is the consequence of elevated de novo lipogenesis, we are looking at the gene important for the de novo lipogenesis. And as expected, normal allele,  relative to normal alleles gene edited and least carrier has more elevated stethotic phenotype. So taken together, at least our organic model  is confirming this GCKR is driving the stethosis, but how does it translate in humans? So that is always the question we are challenged by all of the reviewers.  So we went on looking at the Gilead Science data sets, which is containing like thousands of patient data sets. What we have found out was quite fascinating.  So like I said, our organoid is actually reflecting type two diabetic-like conditions. So we actually stratified NASH patient towards the non-diabetic versus diabetic patients,  each containing like 500-ish patient numbers. But when you look at the non-diabetic ones, this GCKR Liskalo is actually protecting the NAFLD NASH progression.  So lowering the NASH activity score, whereas in diabetic NASH conditions, you are elevating the NASH dependent pathology score, indicating if you sum up all of the population,  you're gonna lose the signal. So that's why we think this was controversial in the past. But the bad news is here, looking at the metformin responses,  which is also controversial for the NASH efficacy, right? So when you look at the non-diabetic patient's population, I'm sorry, diabetic patient's population  with a CC or CT normal allele, you can protect or you can down-regulate the fibrosis activity scores, whereas the Lisk allele carrier  has not been well-responded to the perturbations. And same on the steatopatitis HLO, suggesting the current treatments is not so effective to reverse TT-defined aggressive NASH.  But the good news here is, we look at the mechanistic analysis, so the TT carriers in a NASH-organized model, finding out the mitochondrial dysregulation  is really sitting at the bottom of this damage process. So in collaboration with also Bio, we are able to reverse this phenotype by coupling as well as NAD replenishment approach.  Because of the time, I'm not going in details, but now we are collaborating with the startup companies  to see if we can help out the clinical programs. There are two posters is gonna be presented today. So this is my final slides to conclude. So I think the key takeaways is,  we need to define how much complexity you need, or how simple is complex enough to address your questions, right? So I think simpler model always has an advantage  in terms of reproducibilities or scalabilities. So I think for the people interested in doing a screening, I would definitely suggest to start from the simpler one.  But if you wanna test more very profound, important crosstalk related involving multiple cell types, maybe you're gonna aim for more complicated model system.  So each has a pros and cons. But I think there are significant limitation, at least today. For example, we are completely lacking the system-level crosstalk.  So we don't have immunity systems, we don't have a circuit system, we don't have endocrine system to look at the liver-specific perturbation. So I think the next frontier  would be probably integrating the system in a dish. Or as I said in my middle of talk, maybe transplantation coupled organoid-based studies might be something fill the gap.  So with that, I would like to thank all my colleagues, collaborators, mentors, friends, audience, and ASLD members.  Thank you so much. Thank you.

Video Summary

Dr. Takenori Takebe, an Associate Professor of Pediatrics and the Endowed Chair of Organoid Medicine at Cincinnati Children's Hospital, gave a lecture on how human liver organoids can be used for drug development. Organoids are 3D tissue structures made from human stem cells that mimic the structure and function of organs, allowing for the study of organ development and disease progression. Dr. Takebe's research focuses on creating functional human liver organoids in a petri dish to study liver tissue renewal, development, and disease. He has successfully developed liver organoids that mimic the zonal-specific markers and functionalities of the liver, such as bile acid elimination and cytochrome activities. He has also integrated sinusoidal endothelial cells and demonstrated their role in liver-specific functions such as factor VIII synthesis and immune interactions. Furthermore, Dr. Takebe has used liver organoids to study non-alcoholic fatty liver disease (NAFLD) and has identified genetic variants, such as GCKR, that contribute to the development of NAFLD. He has also shown differences in the response to treatment, such as metformin, based on genetic variants. Looking ahead, Dr. Takebe highlighted the need for more complex models that incorporate multiple cell types and system-level crosstalk, as well as the potential for transplantation-coupled organoid-based studies to bridge the gap between in vitro and in vivo models.

Asset Caption

Presented by Dr. Takanori Takebe, MD, Pediatrics, Cincinnati Children's Hospital Medical Center

The goal of this program is to propose integrative pre-clinical research concept for human NAFLD/NASH, combining patients’ and engineered human tissues using emerging technologies such as induced pluripotency, organoids, gene editing and comprehensive genomics. By introducing population-based organoid system, this investigative strategy may open the door to the studies of pleiotropic factors, potentially transforming the future of precision hepatology.

Keywords

Human liver organoids

Drug development

Liver tissue renewal

Non-alcoholic fatty liver disease

Genetic variants

Treatment response

Complex models