title: Modification of Three Amino Acids in Sodium Taurocholate Cotransporting Polypeptide Renders Mice Susceptible to Infection with Hepatitis D Virus In Vivo.
abstract: Sodium taurocholate cotransporting polypeptide (NTCP) was identified as a functional receptor for hepatitis D virus (HDV) and its helper hepatitis B virus (HBV). In cultured cell lines, HDV infection through mouse NTCP is restricted by residues 84 to 87 of the receptor. This study shows that mice with these three amino acids altered their corresponding human residues (H84R, T86K, and S87N) in endogenous mouse NTCP support de novo HDV infection in vivo HDV infection was documented by the presence of replicative forms of HDV RNA and HDV proteins in liver cells at day 6 after viral inoculation. Monoclonal antibody specifically binding to the motif centered on K86 in NTCP partially inhibited HDV infection. These studies demonstrated specific interaction between the receptor and the viral envelopes in vivo and established a novel mouse model with minimal genetic manipulation for studying HDV infection. The model will also be useful for evaluating entry inhibitors against HDV and its helper HBV.
  
  
  
  NTCP was identified as a functional receptor for both HDV and HBV in cell cultures. We recently showed that neonatal C57BL/6 transgenic (Tg) mice exogenously expressing human NTCP (hNTCP-Tg) in liver support transient HDV infection. In this study, we introduced alterations of three amino acids in the endogenous NTCP of FVB mice through genome editing. The mice with the humanized NTCP residues (H84R, T86K, and S87N) are susceptible to HDV infection, and the infection can be established in both neonatal and adult mice with this editing. We also developed a monoclonal antibody specifically targeting the region of NTCP centered on lysine residue 86, and it can differentiate the modified mouse NTCP from that of the wild type and partially inhibited HDV infection. These studies shed new light on NTCP-mediated HDV infection in vivo, and the NTCP-modified mice provide a useful animal model for studying HDV infection and evaluating antivirals against the infection.
authors.affiliation: Postdoctoral Program, School of Life Sciences, Peking University, Beijing, China National Institute of Biological Sciences, Beijing, China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: He
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Zhiliang
authors.initials: Z
authors.lastname: Cao
authors.affiliation: National Institute of Biological Sciences, Beijing, China Graduate Program, School of Life Sciences, Beijing Normal University, Beijing, China.
authors.forename: Fengfeng
authors.initials: F
authors.lastname: Mao
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Bijie
authors.initials: B
authors.lastname: Ren
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Li
authors.affiliation: National Institute of Biological Sciences, Beijing, China Graduate Program, School of Life Sciences, Tsinghua University, Beijing, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Huiyu
authors.initials: H
authors.lastname: Li
authors.affiliation: National Institute of Biological Sciences, Beijing, China Graduate Program, School of Life Sciences, Peking University, Beijing, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Peng
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Yan
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Yonghe
authors.initials: Y
authors.lastname: Qi
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Yinyan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: National Institute of Biological Sciences, Beijing, China.
authors.forename: Fengchao
authors.initials: F
authors.lastname: Wang
authors.affiliation: National Institute of Biological Sciences, Beijing, China Graduate Program, School of Life Sciences, Tsinghua University, Beijing, China suijianhua@nibs.ac.cn liwenhui@nibs.ac.cn.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Sui
authors.affiliation: National Institute of Biological Sciences, Beijing, China Graduate Program, School of Life Sciences, Beijing Normal University, Beijing, China Graduate Program, School of Life Sciences, Peking University, Beijing, China suijianhua@nibs.ac.cn liwenhui@nibs.ac.cn.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Li
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D004195:Disease Models, Animal; D004198:Disease Susceptibility; D003699:Hepatitis D / Q000821:virology*; D003698:Hepatitis Delta Virus / Q000502:physiology*; D051379:Mice; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D029363:Organic Anion Transporters, Sodium-Dependent / Q000235:genetics* / Q000378:metabolism*; D011991:Receptors, Virus / Q000235:genetics* / Q000378:metabolism*; D027981:Symporters / Q000235:genetics* / Q000378:metabolism*
chemical_list: D050505:Mutant Proteins; D029363:Organic Anion Transporters, Sodium-Dependent; D011991:Receptors, Virus; D027981:Symporters; C071908:sodium-bile acid cotransporter