Functional characterization of organoids derived from irreversibly damaged NASH patient liver.

McCarron, Sarah; Bathon, Brooke; Conlon, Donna M; Abbey, Deepti; Rader, Daniel J; Gawronski, Katerina; Brown, Christopher D; Olthoff, Kim M; Shaked, Abraham; Raabe, Tobias D

BACKGROUND AND AIMS Nonalcoholic steatohepatitis (NASH) will soon become the leading cause of liver transplantation in the US and is also associated with increased COVID-19 mortality. Currently, there are no FDA approved drugs available that slow NASH progression or address NASH liver involvement in COVID-19. Since animal models cannot fully recapitulate human NASH, we hypothesized that stem cells isolated directly from end-stage NASH patient liver may address current knowledge gaps in human NASH pathology. APPROACH AND RESULTS We devised methods allowing derivation, proliferation, hepatic differentiation and extensive characterization of bipotent ductal organoids from irreversibly damaged NASH patient liver. The transcriptomes of organoids derived from NASH liver, but not healthy liver show significant upregulation of pro-inflammatory and cytochrome p450-related pathways, as well as of known liver fibrosis and tumor markers, with the degree of upregulation being NASH patient-specific. Functionally, NASH liver organoids exhibit reduced passaging/growth capacity and show hallmarks of NASH liver, including decreased albumin production, increased free fatty acid induced lipid accumulation, increased sensitivity to apoptotic stimuli and increased CYP450 metabolism. After hepatic differentiation, NASH liver organoids exhibit reduced ability to dedifferentiate back to the biliary state, consistent with the known reduced regenerative ability of NASH livers. Intriguingly, NASH liver organoids also show strongly increased permissiveness to SARS-CoV-2-VSV pseudovirus as well as upregulation of ubiquitin D (Fat10), a known inhibitor of the antiviral interferon host response. CONCLUSION Expansion of primary liver stem cells/organoids derived directly from irreversibly damaged NASH patient liver opens up new experimental avenues for personalized disease modeling and drug development that has the potential to slow human NASH progression and to counteract NASH related SARS-CoV-2 effects.