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Xia, Y. ; Carpentier, A.* ; Cheng, X. ; Block, P.D.* ; Zhao, Y.* ; Zhang, Z.* ; Protzer, U. ; Jake Liang, T.*

Human stem cell-derived hepatocytes as a model for hepatitis B virus infection, spreading and virus-host interactions.

J. Hepatol. 66, 494-503 (2017)

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BACKGROUND AND AIMS: One major obstacle of hepatitis B virus (HBV) research is the lack of efficient cell culture system permissive for viral infection and replication. The aim of our study was to establish a robust HBV infection model by using hepatocyte-like cells (HLCs) derived from human pluripotent stem cells. METHODS: HLCs were differentiated from human embryonic stem cells and induced pluripotent stem cells. Maturation of hepatocyte functions was determined. After HBV infection, viral total DNA, cccDNA, total RNA, pgRNA, HBeAg, HBsAg were measured. RESULTS: More than 90% of the HLCs expressed strong signals of human hepatocyte markers like albumin as well as known host factors required for HBV infection, suggesting that these cells present key features of mature hepatocytes. Notably, HLCs expressed the viral receptor sodium-taurocholate cotransporting polypeptide more stably than primary human hepatocytes (PHHs). HLCs supported robust infection and some spreading of HBV. Finally, by using this model, we identified two host-targeting agents, Genistin and PA452, as novel antivirals. CONCLUSIONS: Stem cells-derived HLCs fully support HBV infection. This novel HBV infection HLCs model offers a unique opportunity to advance our understanding of the molecular details of the HBV life cycle, to further characterize virus-host interactions and to define new targets for HBV curative treatment. LAY SUMMARY: Our study used human pluripotent stem cells to develop hepatocyte-like cells (HLCs) capable of expressing hepatocyte markers and host factors important for HBV infection. These cells fully support HBV infection and virus-host interactions, allowing for the identification of two novel antiviral agents. Thus, stem cell-derived HLCs provide a highly physiologically relevant system to advance our understanding of viral life cycle and provide a new tool for antiviral drug screening and development.

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