Functional Characterization of Organoids Derived From Irreversibly Damaged Liver of Patients With NASH

Abstract

Background and Aims NASH will soon become the leading cause of liver transplantation in the United States and is also associated with increased COVID‐19 mortality. Currently, there are no Food and Drug Administration–approved drugs available that slow NASH progression or address NASH liver involvement in COVID‐19. Because animal models cannot fully recapitulate human NASH, we hypothesized that stem cells isolated directly from end‐stage liver from patients with NASH may address current knowledge gaps in human NASH pathology. Approach and Results We devised methods that allow the derivation, proliferation, hepatic differentiation, and extensive characterization of bipotent ductal organoids from irreversibly damaged liver from patients with NASH. The transcriptomes of organoids derived from NASH liver, but not healthy liver, show significant up‐regulation of proinflammatory and cytochrome p450–related pathways, as well as of known liver fibrosis and tumor markers, with the degree of up‐regulation being patient‐specific. Functionally, NASH liver organoids exhibit reduced passaging/growth capacity and hallmarks of NASH liver, including decreased albumin production, increased free fatty acid–induced lipid accumulation, increased sensitivity to apoptotic stimuli, and increased cytochrome P450 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 severe acute respiratory syndrome–coronavirus 2 (SARS‐CoV‐2) vesicular stomatitis pseudovirus as well as up‐regulation of ubiquitin D, a known inhibitor of the antiviral interferon host response. Conclusion Expansion of primary liver stem cells/organoids derived directly from irreversibly damaged liver from patients with NASH opens up 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.

SARS-CoV-2 infection and liver involvement

“…Therefore, a potential mechanism for exaggerated SARS-CoV-2 liver injury in underlying CLD is due to greater burden of hepatocyte infection and consequently widespread hepatocyte cell death. An elegant demonstration of this is found in liver-derived organoids from cirrhotic NASH patients, which show markedly increased permissiveness to SARS-CoV-2 infection and pro-inflammatory gene expression compared to liver organoids from non-cirrhotic donors [ 12 ].…”

Organoid Models of SARS-CoV-2 Infection: What Have We Learned about COVID-19?

“…McCarron and colleagues [119] took organoid technology one step further. Nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) are becoming recognized as co-morbidities of COVID-19 [120].…”

“…Infection with the SARS-CoV-2-VSV pseudovirus that expresses the SARS-CoV-2 S protein and requires ACE2 and TMPRSS2 for fusion with the cellular membrane was more efficient in NASH organoids than in organoids established from healthy tissues. Interestingly, increased permissiveness to SARS-CoV-2 was not associated with elevated ACE2 or TMPRSS2 expression but an upregulation of ubiquitin D, a known inhibitor of the interferon antiviral response, in the NASH organoids akin to human NASH liver tissue [119]. Establishing organoids from diseased tissues has the potential to reveal the mechanisms of comorbidities in COVID-19.…”

In vitro models for non-alcoholic fatty liver disease: Emerging platforms and their applications

“…(2019) , treatment with FFA induced a dose-dependent accumulation of intracellular lipids and establishment of a NASH phenotype. McCarron et al. (2021) instead developed bipotent ductal organoids derived from end-stage NASH patient liver explants, with normal donor livers as controls.…”

“…Several organoid models recapitulating NAFLD phenotypes have used bulk RNA sequencing (RNA-seq) to identify gene signatures and pathways associated with the disease ( McCarron et al., 2021 ; Elbadawy et al., 2020 ; Beisner et al., 2021 ; Ouchi et al., 2019 ). Yet, only a few have included flow cytometry ( Ouchi et al., 2019 ; Gurevich et al., 2020 ; Ramli et al., 2020 ; Abbey et al., 2020 ) or single-cell RNA-seq ( Ouchi et al., 2019 ) in their research to determine the proportion of the different cell types present in these organoids.…”