Organoid-guided Precision Hepatology for Steatohepatitis

PROJECT SUMMARY Non-alcoholic fatty liver disease (NAFLD) affects billions of people. NAFLD was first defined in the USA 35 years ago, yet progress toward a true cure has been incremental and limited to lifestyle modification. The long-term goal of our research program is to take a fresh view on human liver metabolism through the lens of precision hepatology to define personalized mechanisms as a fundamental step toward developing evidence-based, rational solutions for mechanism-directed diagnostic and therapeutic investigation. Here, we propose to explore liver effect by integrative pre-clinical research, combining patients’ and engineered human tissues using new technologies such as human organoids, gene editing and comprehensive genomics/transcriptomics. We propose to use our pioneering steatohepatitis organoids in combination with an innovative population-scale pluripotent stem cell (PSC) approach to identifying NASH genotype-phenotype associations under disrupted metabolic states. Our Preliminary Data using this human organoid GWAS system suggests that GCKR rs1260326 confer risk for developing steatosis and inflammation under oleic acid induced insulin resistant conditions, supported by cohort data from thousands of NASH patients. Based on these exciting Preliminary Data, the overall objective in this application is to elucidate variant-dependent effects of common coding single nucleotide polymorphism on liver specific steatosis and inflammation under disrupted metabolic states. Our central hypothesis is that GCKR rs1260326 activates glucokinase to trigger hepatic lipogenesis under an insulin resistant state, thereby exacerbating pathogenic inflammation through mitochondrial oxidative stress. The rationale for the proposed research is that a mechanistic determination of GCKR genetic predisposition in T2D complicated steatohepatitis will provide a strong foundation for identifying susceptible populations and molecular pathway directed therapy development. We will critically test and validate our hypothesis in the following Aims: In Aim1, we will determine the mechanistic role of GCKR rs1260326 in steatohepatitis liver organoids. In Aim2, we will interrogate GCKR rs1260326 mediated mitochondrial dysregulation in organoid and patients. In Aim3, we will redefine GCKR rs1260326 and steatohepatitis associations in diabetic NASH patients. At the completion, we will have determined how common risk variants and metabolic activities in the liver interact to cause aggressive forms of NASH, informing risk stratification strategies, as well as the potentially diverging efficacy of available therapies. Our studies will lead to a better mechanistic understanding of mitochondrial dysregulation associated with GCKR rs1260326 at the level of liver, forming a basis for mitochondrial metabolism-directed therapeutic investigations. Further, by demonstrating the utility of our population-based organoid model system, this investigative strategy may open the door to many other studies of pleiotropic heritable factors, potentially transforming the future of precision hepatology.