Development and characterization of a human skin stem cell-derived in vitro liver model for anti-NASH drug screening.

Non-alcoholic fatty liver disease (NAFLD) ranges from benign steatosis to severe, life-threatening non-alcoholic steatohepatitis (NASH). Steatosis is mostly asymptomatic and does not cause health complications. However, in 5-10% of the cases it proceeds to NASH in which hepatic inflammation occurs. Today’s investigations of NAFLD and NASH rely mainly on animal models, which are not representative for the human situation. Therefore, the research objective of this project is to develop a predictive, human-based in vitro model that could be used to study NASH, identify druggable targets against it and evaluate potential novel anti-NASH compounds. The host laboratory has previously introduced a human stem cell-based hepatic model that could successfully predict drug-induced liver steatosis. This model will now be further developed to mimic NASH in vitro. Hereby, hepatic cells will be first exposed to steatogenic compounds, in order to obtain steatosis. Subsequently, inflammation will be introduced by adding a cytokine cocktail and by co-culture with immune cells that secrete the inflammatory factors. Characterization of the novel model and comparison with samples obtained from NASH patients will be done using RT-qPCR, state-of-the-art transcriptomics and specific functional assays for the determination of lipid accumulation, mitochondrial stress, apoptosis and cytokine secretion. The applicability of the model for drug discovery will be assessed by testing anti-NASH compounds.