title: Human stem cell-derived hepatocytes as a model for hepatitis B virus infection, spreading and virus-host interactions.
abstract: 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.
  
  
  
  HLCs were differentiated from human embryonic stem cells and induced pluripotent stem cells. Maturation of hepatocyte functions was determined. After HBV infection, total viral DNA, cccDNA, total viral RNA, pgRNA, HBeAg and HBsAg were measured.
  
  
  
  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 possessed 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.
  
  
  
  Stem cell-derived HLCs fully support HBV infection. This novel HLC HBV infection 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.
  
  
  
  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.
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Yuchen
authors.initials: Y
authors.lastname: Xia
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Carpentier
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Cheng
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Peter Daniel
authors.initials: PD
authors.lastname: Block
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Yao
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Zhensheng
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Institute of Virology, Technical University of Munich/Helmholtz Zentrum München, 81675 Munich, Germany; German Center for Infection research (DZIF), Munich partner site, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Protzer
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States. Electronic address: jakel@bdg10.niddk.nih.gov.
authors.forename: T Jake
authors.initials: TJ
authors.lastname: Liang
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D055808:Drug Discovery; D053595:Embryonic Stem Cells / Q000166:cytology / Q000821:virology; D006509:Hepatitis B / Q000188:drug therapy / Q000473:pathology / Q000821:virology*; D006515:Hepatitis B virus / Q000472:pathogenicity / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology / Q000821:virology*; D054884:Host-Pathogen Interactions; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000821:virology; D008954:Models, Biological; D013234:Stem Cells / Q000166:cytology / Q000821:virology*; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents
keywords: Antiviral agents; Antivirals; DNA; Hepatitis B virus; Hepatocytes; Model; Stem cell; Viral