Patient transcriptomic based in vivo functional genetic dissection of NAFLD for therapeutic target identification

Abstract

Chronic liver disease is a leading cause of death globally and non-alcoholic fatty liver disease (NAFLD) is put in the spotlight with increasing prevalence. With no effective therapeutics apart from lifestyle modifications and liver transplantation, treatment of NAFLD remains a challenge. As NAFLD is characterised by impaired liver regeneration capacity which causes eventual liver failure, enhancing hepatocyte proliferation may ameliorate disease progression. Here, we identified 6 novel gene candidates potentially involved in regulating liver regeneration, through in vivo RNA interference functional genetic screen using small hairpin RNA (shRNA). The effect of shRNA-mediated knockdown of the candidates on cell proliferation were evaluated in vitro. Based on the results, we narrowed down to one candidate, Target 1. Downregulation of Target 1 increased hepatocyte proliferation in vitro and in vivo. Similar phenotype was also observed in hepatocytes treated with Target 1 antagonist. Our results suggest that Target 1 may be further explored for NAFLD treatment. We also established an in vitro steatosis model with NAFLD phenotypes – excessive lipid accumulation and slower cell proliferation. Future studies can be focused on elucidating mechanism of Target 1 using the steatosis model, to gain insights into pathways regulating liver regeneration and to identify new therapeutic targets.