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.