Human liver organoids; a patient-derived primary model for HBV Infection and Related Hepatocellular Carcinoma

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 interrogate the potential of human liver organoids as a platform for modeling HBV infection and related tumorigenesis. We show that organoids derived from HBV-infected patients display an aberrant early cancer gene signature, which clusters with the HCC cohort on the TCGA LIHC dataset and away from healthy liver tissue. Furthermore, we demonstrate HBV infection in healthy donor liver organoids after challenge with recombinant virus or HBV infected patient serum. Ex vivo infected liver organoids produced cccDNA, expressed intracellular HBV RNA and proteins, and produced infectious HBV. HBV replication supported by ex vivo infected liver organoids was blocked by treatment with Tenofovir, highlighting the potential of this model system as a primary differentiated hepatocyte platform for HBV drug screening. Interestingly, transgenic organoids exogenously over expressing the HBV receptor NTCP by lentiviral transduction are not more susceptible to HBV, suggesting the necessity for additional host factors for efficient infection. Finally, we generated transgenic organoids harboring integrated HBV, representing a long-term culture system also suitable for viral production and the study of HBV transcription.