The molecular events that drive Hepatitis B virus (HBV)-mediated transformation and tumorigenesis have remained largely unclear, due to the absence of a relevant primary model system. Here we propose the use of human liver organoids as a platform for modeling HBV infection and related tumorigenesis. We first describe a primary ex vivo HBV-infection model derived from healthy donor liver organoids after challenge with recombinant virus or HBV-infected patient serum. HBV infected organoids produced cccDNA, HBeAg, expressed intracellular HBV RNA and proteins, and produced infectious HBV. This ex vivo HBV infected primary differentiated hepatocyte organoid platform was amenable to drug screening for both anti-HBV activity as well as for drug-induced toxicity. We also studied HBV replication in transgenically modified organoids; liver organoids exogenously overexpressing the HBV receptor NTCP after lentiviral transduction were not more susceptible to HBV, suggesting the necessity for additional host factors for efficient infection. We also generated transgenic organoids harboring integrated HBV, representing a long-term culture system also suitable for viral production and the study of HBV transcription. Finally, we generated HBV-infected patient-derived liver organoids from non-tumor cirrhotic tissue of explants from liver transplant patients. Interestingly, transcriptomic analysis of patient-derived liver organoids indicated the presence of an aberrant early cancer gene signature, which clustered with the HCC cohort on the TCGA LIHC dataset and away from healthy liver tissue, and may provide invaluable novel biomarkers for the development of HCC and surveillance in HBV infected patients.© 2021, De Crignis et al.
About The Expert
Elisa De Crignis
Tanvir Hossain
Shahla Romal
Fabrizia Carofiglio
Panagiotis Moulos
Mir Mubashir Khalid
Shringar Rao
Ameneh Bazrafshan
Monique Ma Verstegen
Farzin Pourfarzad
Christina Koutsothanassis
Helmuth Gehart
Tsung Wai Kan
Robert-Jan Palstra
Charles Boucher
Jan Mn IJzermans
Meritxell Huch
Sylvia F Boj
Robert Vries
Hans Clevers
Luc Jw van der Laan
Pantelis Hatzis
Tokameh Mahmoudi
References
PubMed