DEVELOPMENT OF IN VITRO NON-ALCOHOLIC STEATOHEPATITIS (NASH) MODEL FOR DRUG TESTING AND PATHOGENESIS STUDIES

Abstract

Non-alcoholic steatohepatitis (NASH) is a severe chronic fatty liver disease that currently has no effective intervention. Hepatocellular ballooning is one of the key features of NASH diagnosis. Due to their scarcity, it is difficult to study in vivo and isolate for in vitro investigations. How ballooned hepatocytes form and its role in NASH progression is poorly understood. Here, an in vitro hepatocellular ballooning model is developed using HepaRG cells subjected to free fatty acids and Kupffer cell condition media to induce NASH-like phenotype. The induced ballooned hepatocytes exhibited typical features observed in in vivo settings. We recognized cytokeratin 18 fragment (M30) as a quantifiable biomarker for hepatocellular ballooning, which positively correlated with the level of steatosis and inflammation. ScRNA-seq was performed to find the ballooned hepatocyte cluster. An AMPK signalling was proposed to be involved in formation mechanism. Lastly, the application for scalable high-throughput drug screening has been demonstrated.