Modeling of the hepatitis B virus life cycle and the efficacy of antivirals in human iPSC-derived hepatic organoids

Hepatitis B virus (HBV) infection remains a major global health burden, affecting approximately 296 million people worldwide, and yet progress in mechanistic studies and development of antivirals has been limited by the lack of physiologically relevant and sustainable in vitro models. This study established a human induced pluripotent stem cell (hiPSC)-derived multilineage hepatic organoid system that robustly supports the complete HBV life cycle, including viral entry, replication, covalently closed circular DNA (cccDNA) formation, antigen secretion, and production of infectious progeny virus. These organoids exhibit stable expression of sodium taurocholate cotransporting polypeptide (NTCP), a key receptor for HBV entry, and remain viable long term under infection conditions for at least 20 days, with sustained secretion of HBsAg and HBeAg. Importantly, the model recreates key pathological features of chronic HBV infection, including downregulation of hepatocyte functional genes (e.g., ALB and CYP3A4) and induction of fibrosis-associated markers such as COL1A1, reflecting early extracellular matrix remodeling. Moreover, results indicated the utility of this platform in the evaluation of antivirals. Treatment with tenofovir effectively reduced viral DNA and antigen production without affecting cccDNA levels, whereas bulevirtide resulted in stage-specific inhibition of viral entry, highlighting the model's capacity to resolve mechanism-of-action differences. At the same time, drug-induced hepatotoxicity was assessed within the same system. Collectively, this hiPSC-derived hepatic organoid model provides a scalable and physiologically relevant platform that bridges the gap between conventional cell culture and in vivo systems, offering a powerful tool for studying HBV pathogenesis, host-virus interactions, and preclinical antiviral discovery.