PT  - JOURNAL ARTICLE
AU  - Elisa De Crignis
AU  - Shahla Romal
AU  - Fabrizia Carofiglio
AU  - Panagiotis Moulos
AU  - Monique M.A. Verstegen
AU  - Mir Mubashir Khalid
AU  - Farzin Pourfarzad
AU  - Shringar Rao
AU  - Ameneh Bazrafshan
AU  - Christina Koutsothanassis
AU  - Helmuth Gehart
AU  - Tsung Wai Kan
AU  - Robert-Jan Palstra
AU  - Charles Boucher
AU  - Jan M.N. IJzermans
AU  - Meritxell Huch
AU  - Sylvia F. Boj
AU  - Robert Vries
AU  - Hans Clevers
AU  - Luc van der Laan
AU  - Pantelis Hatzis
AU  - Tokameh Mahmoudi
TI  - Human liver organoids; a patient-derived primary model for HBV Infection and Related Hepatocellular Carcinoma
AID  - 10.1101/568147
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 568147
4099  - http://biorxiv.org/content/early/2020/11/11/568147.short
4100  - http://biorxiv.org/content/early/2020/11/11/568147.full
AB  - 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, 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 by 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 disease surveillance and development of HCC in HBV infected patients.Competing Interest StatementThe authors have declared no competing interest.