Development of a cell-based assay to identify hepatitis B virus entry inhibitors targeting the sodium taurocholate cotransporting polypeptide
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Kei Miyakawa1, Satoko Matsunaga1, Yutaro Yamaoka1,2, Mina Dairaku1, Kento Fukano3, Hirokazu Kimura4, Tomoyuki Chimuro2, Hironori Nishitsuji5, Koichi Watashi3, Kunitada Shimotohno5, Takaji Wakita3 and Akihide Ryo1
1Department of Microbiology, Yokohama City University School of Medicine, Kanagawa 236-0004, Japan
2Isehara Research Laboratory, Technology and Development Division, Kanto Chemical Co., Inc., Kanagawa 259-1146, Japan
3Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan
4School of Medical Technology, Faculty of Health Sciences, Gunma Paz University, Gunma 370-0006, Japan
5Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Chiba 272-8516, Japan
Akihide Ryo, email: [email protected]
Keywords: HBV-permissive cell; anti-NTCP monoclonal antibody; glabridin; innate immune signaling; ISG
Received: January 06, 2018 Accepted: April 24, 2018 Published: May 04, 2018
Sodium taurocholate cotransporting polypeptide (NTCP) is a major entry receptor of hepatitis B virus (HBV) and one of the most attractive targets for anti-HBV drugs. We developed a cell-mediated drug screening method to monitor NTCP expression on the cell surface by generating a HepG2 cell line with tetracycline-inducible expression of NTCP and a monoclonal antibody that specifically detects cell-surface NTCP. Using this system, we screened a small molecule library for compounds that protected against HBV infection by targeting NTCP. We found that glabridin, a licorice-derived isoflavane, could suppress viral infection by inducing caveolar endocytosis of cell-surface NTCP with an IC50 of ~40 μM. We also found that glabridin could attenuate the inhibitory effect of taurocholate on type I interferon signaling by depleting the level of cell-surface NTCP. These results demonstrate that our screening system could be a powerful tool for discovering drugs targeting HBV entry.
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