Metabolism-mediated epigenetic regulation of hepatic stellate cell activation during liver fibrosis

The National Institute of Diabetes and Digestive and Kidney Diseases awarded Mayo Clinic $149,190 on September 30, 2026, under the Diabetes, Digestive, and Kidney Diseases Extramural Research program (CFDA 93.847) to conduct research on metabolism-mediated epigenetic regulation of hepatic stellate cell activation during liver fibrosis.

The funded research investigates novel mechanisms driving liver fibrosis progression to guide development of potential treatment strategies. The project tests the central hypothesis that glycolysis-mediated histone 3 lysine 9 acetylation (H3K9AC) by lysine acetyltransferase 2B (KAT2B) promotes hepatic stellate cell activation and liver fibrosis. Work employs sophisticated cellular and animal models, including in vitro carbon-13 glucose tracing, chromatin immunoprecipitation sequencing, dCas-KRAB models, in vivo HSC-specific KAT2B deletion models, and ex vivo liver fibrosis models using human precision-cut liver slices. Preliminary findings demonstrate that platelet-derived growth factor increases acetyl-CoA levels and KAT2B expression in hepatic stellate cells, that PDGF-mediated glycolysis increases H3K9AC, and that pharmacological inhibition of H3K9AC by CPTH6 prevents hepatic stellate cell proliferation and migration in vitro and reduces liver fibrosis in vivo.

Mayo Clinic is the recipient organization. The award is structured as a Project Grant with a period of performance from September 30, 2026, through March 30, 2029, and performance takes place in Rochester, Minnesota.

Abstract

Cirrhotic stage liver disease is the 9TH leading cause of mortality in the US, and no treatment exists for late-stage disease other than liver transplantation. Thus, the overall objective of this proposal is to elucidate novel mechanisms that drive liver fibrosis progression and to guide the development of potential treatment strategies. Liver fibrosis is characterized by the activation of hepatic stellate cells (hscs). Our preliminary data in primary human and mouse hscs as well as in vivo demonstrate that 1. Platelet derived growth factor (pdgf) increases acetyl-COA levels in hscs and the expression of one of the histone acetyltransferases, lysine acetyltransferase 2B (KAT2B); 2. Pdgf-mediated glycolysis increases the epigenetic transcription activation mark, histone 3 lysine 9 acetylation (H3K9AC) in hscs; 3. In vitro pharmacological inhibition of H3K9AC by CPTH6 prevents proliferation and migration of pdgf-stimulated hscs; 4. In vivo drug inhibition of H3K9AC by CPTH6 reduces liver fibrosis; 5. H3K9AC levels and KAT2B expression are increased in mice fibrotic livers. We utilized our novel findings to generate the cental hypothesis of the current proposal that glycolysis-mediated H3K9AC by KAT2B promotes HSC activation and liver fibrosis. We will employ sophisticated cellular and animal models, including in vitro carbon-13 glucose tracing, chromatin immunoprecipitation sequencing (chip-SEQ), in vitro utilization of dcas-krab model, in vivo HSC-specific KAT2B deletion model as well as an EX vivo liver fibrosis model with human precision cut liver slices (pcls), to investigate the following integrated, yet independent aims. In aim 1, we will test the hypothesis that glycolysis derived acetyl-COA promotes H3K9AC dependent pro- fibrogenic gene expression. We will uncover the gene programs regulated by glycolysis- linked H3K9AC in hscs by: a. Studying how glucose metabolism contributes to the production of acetyl-COA, the main metabolite to acetylate the histones, and how this affects H3K9AC; and b. Investigating how investigating the gene promoters in pdgf-stimulated hscs that are enriched with H3K9AC. In aim 2, we will test the hypothesis that KAT2B- dependent H3K9AC and subsequent HSC activation induce liver fibrosis. We will dissect how HSC-specific H3K9AC amplifies liver fibrosis by: a. Studying how KAT2B, the acetyltransferase responsible for H3K9AC, impacts HSC activation in vitro, including proliferation and migration; and b. Examining how H3K9AC amplifies EX vivo and in vivo liver fibrosis. This novel and innovative line of inquiry will define an HSC-specific glycolysis- dependent model of liver fibrosis amplification and set a trajectory towards new and significant advances to treat liver fibrosis and cirrhosis in humans. Lastly, completing the planned education outlined in goals for fellowship training and this project will contribute to my overall formation as an independent scientist with a career in advancing the research for digestive health.