title: Stem cell-derived hepatocytes: A novel model for hepatitis E virus replication.
abstract: OBJECTIVE
  Yearly, approximately 20million people become infected with the hepatitis E virus (HEV) resulting in over 3million cases of acute hepatitis. Although HEV-mediated hepatitis is usually self-limiting, severe cases of fulminant hepatitis as well as chronic infections have been reported, resulting annually in an estimated 60,000 deaths. We studied whether pluripotent stem cell (PSC)-derived hepatocytes, mesodermal and/or neuroprogenitor cells support HEV replication.
  
  
  METHODS
  Human PSC were differentiated towards hepatocyte-like cells, mesodermal cells and neuroprogenitors and subsequently infected with HEV. Infection and replication of HEV was analyzed by qRT-PCR, RNA in situ hybridization, negative strand RT-PCR, production of infectious virions and transfection with a transient HEV reporter replicon.
  
  
  RESULTS
  PSC-derived hepatocytes supported the complete replication cycle of HEV, as demonstrated by the intracellular presence of positive and negative strand HEV RNA and the production of infectious virions. The replication of the virus in these cells was inhibited by the antiviral drugs ribavirin and interferon-α2b. In contrast to PSC-derived hepatocytes, PSC-derived mesodermal cells and neuroprogenitors only supported HEV replication upon transfection with a HEV subgenomic replicon.
  
  
  CONCLUSIONS
  We demonstrate that PSC can be used to study the hepatotropism of HEV infection. The complete replication cycle of HEV can be recapitulated in infected PSC-derived hepatocytes. By contrast other germ layer cells support intracellular replication but are not infectable with HEV. Thus the early steps in the viral cycle are the main determinant governing HEV tissue tropism. PSC-hepatocytes offer a physiological relevant tool to study the biology of HEV infection and replication and may aid in the design of therapeutic strategies.
authors.affiliation: Stem Cell Institute, University of Leuven (KU Leuven), Leuven, Belgium. Electronic address: nicky.helsen@med.kuleuven.be.
authors.forename: Nicky
authors.initials: N
authors.lastname: Helsen
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, University of Leuven (KU Leuven), Belgium.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Debing
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, University of Leuven (KU Leuven), Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Paeshuyse
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, University of Leuven (KU Leuven), Belgium.
authors.forename: Kai
authors.initials: K
authors.lastname: Dallmeier
authors.affiliation: Stem Cell Institute, University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Ruben
authors.initials: R
authors.lastname: Boon
authors.affiliation: Institut d'Investigacions Biomèdiques August Pi I Sunyer (IDIBAPS) - Hospital Clinic, Liver Unit, Spain.
authors.forename: Mar
authors.initials: M
authors.lastname: Coll
authors.affiliation: Institut d'Investigacions Biomèdiques August Pi I Sunyer (IDIBAPS) - Hospital Clinic, Liver Unit, Spain.
authors.forename: Pau
authors.initials: P
authors.lastname: Sancho-Bru
authors.affiliation: Stem Cell Institute, University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Christel
authors.initials: C
authors.lastname: Claes
authors.affiliation: Laboratory of Virology and Experimental Chemotherapy, Rega Institute for Medical Research, University of Leuven (KU Leuven), Belgium.
authors.forename: Johan
authors.initials: J
authors.lastname: Neyts
authors.affiliation: Stem Cell Institute, University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Verfaillie
mesh_terms: D056945:Hep G2 Cells; D016752:Hepatitis E virus / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology*; D012367:RNA, Viral / Q000032:analysis; D053586:Virus Internalization; D014779:Virus Replication
chemical_list: D012367:RNA, Viral
keywords: Cell culture model; Hepatitis E virus; Hepatocytes; Pluripotent stem cells