title: Dysregulation of retinoic acid receptor diminishes hepatocyte permissiveness to hepatitis B virus infection through modulation of sodium taurocholate cotransporting polypeptide (NTCP) expression.
abstract: Sodium taurocholate cotransporting polypeptide (NTCP) is an entry receptor for hepatitis B virus (HBV) and is regarded as one of the determinants that confer HBV permissiveness to host cells. However, how host factors regulate the ability of NTCP to support HBV infection is largely unknown. We aimed to identify the host signaling that regulated NTCP expression and thereby permissiveness to HBV. Here, a cell-based chemical screening method identified that Ro41-5253 decreased host susceptibility to HBV infection. Pretreatment with Ro41-5253 inhibited the viral entry process without affecting HBV replication. Intriguingly, Ro41-5253 reduced expression of both NTCP mRNA and protein. We found that retinoic acid receptor (RAR) regulated the promoter activity of the human NTCP (hNTCP) gene and that Ro41-5253 repressed the hNTCP promoter by antagonizing RAR. RAR recruited to the hNTCP promoter region, and nucleotides -112 to -96 of the hNTCP was suggested to be critical for RAR-mediated transcriptional activation. HBV susceptibility was decreased in pharmacologically RAR-inactivated cells. CD2665 showed a stronger anti-HBV potential and disrupted the spread of HBV infection that was achieved by continuous reproduction of the whole HBV life cycle. In addition, this mechanism was significant for drug development, as antagonization of RAR blocked infection of multiple HBV genotypes and also a clinically relevant HBV mutant that was resistant to nucleoside analogs. Thus, RAR is crucial for regulating NTCP expression that determines permissiveness to HBV infection. This is the first demonstration showing host regulation of NTCP to support HBV infection.
authors.affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan, the Micro-signaling Regulation Technology Unit, RIKEN Center for Life Science Technologies, Wako 351-0198, Japan.
authors.forename: Senko
authors.initials: S
authors.lastname: Tsukuda
authors.affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan, kwatashi@nih.go.jp.
authors.forename: Koichi
authors.initials: K
authors.lastname: Watashi
authors.affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Iwamoto
authors.affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Suzuki
authors.affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Aizaki
authors.affiliation: the Department of Translational Oncology, St. Marianna University School of Medicine, Kawasaki 216-8511, Japan.
authors.forename: Maiko
authors.initials: M
authors.lastname: Okada
authors.affiliation: the Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa 272-8516, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Sugiyama
authors.affiliation: the Micro-signaling Regulation Technology Unit, RIKEN Center for Life Science Technologies, Wako 351-0198, Japan.
authors.forename: Soichi
authors.initials: S
authors.lastname: Kojima
authors.affiliation: the Department of Virology and Liver Unit, Nagoya City University Graduate School of Medicinal Sciences, Nagoya 467-8601, Japan, and.
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: the Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikawa 272-8516, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Mizokami
authors.affiliation: the Liver Research Center Rhode Island Hospital, Warren Alpert School of Medicine, Brown University, Providence, Rhode Island 02912.
authors.forename: Jisu
authors.initials: J
authors.lastname: Li
authors.affiliation: the Liver Research Center Rhode Island Hospital, Warren Alpert School of Medicine, Brown University, Providence, Rhode Island 02912.
authors.forename: Shuping
authors.initials: S
authors.lastname: Tong
authors.affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology; D001565:Benzoates / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002839:Chromans / Q000737:chemistry / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D005838:Genotype; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000821:virology*; D054884:Host-Pathogen Interactions / Q000187:drug effects; D006801:Humans; D015151:Immunoblotting; D015394:Molecular Structure; D029363:Organic Anion Transporters, Sodium-Dependent / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D018168:Receptors, Retinoic Acid / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D027981:Symporters / Q000235:genetics / Q000378:metabolism*
chemical_list: D000998:Antiviral Agents; D001565:Benzoates; C000597561:CD2665; D002839:Chromans; D029363:Organic Anion Transporters, Sodium-Dependent; D018168:Receptors, Retinoic Acid; D027981:Symporters; C084904:Ro 41-5253; C071908:sodium-bile acid cotransporter; D000218:Adamantane
keywords: Chemical Biology; HBV; Hepatitis Virus; Infection; NTCP; Permissive; RAR; Retinoid; Transcription; Transporter