Identification of host factors that regulate host susceptibility to hepatitis B virus infection and analysis of mechanisms.

• Project/Area Number: 16K19145
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Virology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Tsukuda Senko 国立研究開発法人理化学研究所, 生命医科学研究センター, 研究員 (20710263)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: B型肝炎 / B型肝炎ウイルス / 受容体 / NTCP / 宿主因子 / 局在制御 / ウイルス / HBV / ウィルス / 感染症

Outline of Final Research Achievements

Recent identification of NTCP as a receptor for hepatitis B virus (HBV) has facilitated the analysis of molecular mechanism of the HBV entry, however, the mechanism has not been fully understood. In this study, we aimed to identify host factors that regulate host susceptibility to HBV infection and analyze mechanisms. We identified host factors that affected on the HBV entry process through a transcriptome analysis of both HBV susceptible and non-susceptible cells. Further analysis revealed that one of the identified factors was involved in the HBV internalization into the cells through the regulation of NTCP localization in the cytosol without affecting NTCP expression. These findings will promote understanding of the mechanism of the HBV entry process after the viral attachment of its receptor.

Academic Significance and Societal Importance of the Research Achievements

B型肝炎ウイルス研究では、汎用的かつ高効率にHBV感染を許容する感染培養細胞系の改善、およびマウスモデルなどのin vivo評価系の樹立が求められている。近年HBV感染受容体としてNTCPが同定され解析が進んだが、HBV侵入過程の詳細な制御機構に関しては未だほとんど不明である。今後さらに詳細な解析が必要であるが、本研究で同定したHBVの細胞内への侵入過程を制御する宿主因子は、HBV感染の動物モデルのあるいは新たな抗HBV薬の標的となり得るため、新たな薬剤開発に寄与できると考えられる。

Research Products

• [Journal Article] Troglitazone Impedes the Oligomerization of Sodium Taurocholate Cotransporting Polypeptide and Entry of Hepatitis B Virus Into Hepatocytes (2019)
  • Author(s): Fukano Kento、Tsukuda Senko、Oshima Mizuki、Suzuki Ryosuke、Aizaki Hideki、Ohki Mio、Park Sam-Yong、Muramatsu Masamichi、Wakita Takaji、Sureau Camille、Ogasawara Yuki、Watashi Koichi
  • Journal Title: Frontiers in Microbiology Volume: 9 Pages: 3257-3257
  • DOI: 10.3389/fmicb.2018.03257
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Concept of Viral Inhibitors via NTCP. (2019)
  • Author(s): 1)Tsukuda S, Fukano K, Watashi K, Wakita T.
  • Journal Title: Semin Liver Dis. Volume: 39 Issue: 01 Pages: 78-95
  • DOI: 10.1055/s-0038-1676804
  • Peer Reviewed
• [Journal Article] A Single Adaptive Mutation in Sodium Taurocholate Cotransporting Polypeptide Induced by Hepadnaviruses Determines Virus Species Specificity (2018)
  • Author(s): Takeuchi Junko S.、Fukano Kento、Iwamoto Masashi、Tsukuda Senko、Suzuki Ryosuke、Aizaki Hideki、Muramatsu Masamichi、Wakita Takaji、Sureau Camille、Watashi Koichi
  • Journal Title: Journal of Virology Volume: 93 Issue: 5 Pages: 04132-18
  • DOI: 10.1128/jvi.01432-18
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A new strategy to identify hepatitis B virus entry inhibitors by AlphaScreen technology targeting the envelope-receptor interaction. (2018)
  • Author(s): Saso W, Tsukuda S, Ohashi H, Fukano K, Morishita R, Matsunaga S, Ohki M, Ryo A, Park SY, Suzuki R, Aizaki H, Muramatsu M, Sureau C, Wakita T, Matano T, Watashi K.
  • Journal Title: Biochem Biophys Res Commun Volume: 印刷中 Issue: 2 Pages: 374-379
  • DOI: 10.1016/j.bbrc.2018.04.187
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chemical array system, a platform to identify novel hepatitis B virus entry inhibitors targeting sodium taurocholate cotransporting polypeptide (2018)
  • Author(s): Kaneko M, Futamura Y, Tsukuda S, Kondoh Y, Sekine T, Hirano H, Fukano K, Ohashi H, Saso W, Morishita R, Matsunaga S, Kawai F, Ryo A, Park S-Y, Suzuki R, Aizaki H, Ohtani N, Sureau C, Wakita T, Osada H, Watashi K
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 2769-2769
  • DOI: 10.1038/s41598-018-20987-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cyclosporin derivatives inhibit hepatitis B virus entry without interfering with NTCP transporter activity. (2017)
  • Author(s): Shimura S, Watashi K, Fukano K, Peel M, Sluder A, Kawai F, Iwamoto M, Tsukuda S, Takeuchi S J, Miyake T, Sugiyama M, Ogasawara Y, Park SY, Tanaka Y, Kusuhara H, Mizokami M, Sureau C, Wakita T
  • Journal Title: J Hepatol Volume: 66 Issue: 4 Pages: 685-692
  • DOI: 10.1016/j.jhep.2016.11.009
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A new class of hepatitis B and D virus entry inhibitors, proanthocyanidin and its analogs, that directly act on the viral large surface proteins. (2017)
  • Author(s): Tsukuda S, Watashi K, Hojima T, Isogawa M, Iwamoto M, Omagari K, Suzuki R, Aizaki H, Kojima S, Sugiyama M, Saito A, Tanaka Y, Mizokami M, Sureau C, Wakita T
  • Journal Title: Hepatology Volume: 65 Issue: 4 Pages: 1104-1116
  • DOI: 10.1002/hep.28952
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Fungus-Derived Neoechinulin B as a Novel Antagonist of Liver X Receptor, Identified by Chemical Genetics Using a Hepatitis C Virus Cell Culture System (2016)
  • Author(s): Nakajima S, Watashi K, Ohashi H, Kamisuki S, Izaguirre-Carbonell J, Kwon AT, Suzuki H, Kataoka M, Tsukuda S, Okada M, Moi ML, Takeuchi T, Arita M, Suzuki R, Aizaki H, Kato T, Suzuki T, Hasegawa H, Takasaki T, Sugawara F, Wakita T
  • Journal Title: J Virol Volume: 90(20) Issue: 20 Pages: 9058-74
  • DOI: 10.1128/jvi.00856-16
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Oligomerization of NTCP induces hepatitis B virus internalization, (2018)
  • Author(s): Kento Fukano, Senko Tsukuda, Mio Ohki, Sam-Yong Park, Ryosuke Suzuki, Hideki Aizaki, Masamichi Muramatsu, Takaji Wakita, Yuki Ogasawara, Koichi Watashi,
  • Organizer: International HBV Meeting
  • Int'l Joint Research
• [Presentation] 感染培養系を用いたHBV創薬と侵入機構の解析 (2017)
  • Author(s): 九十田千子
  • Organizer: 第13回肝免疫・ウイルス・フロンティア（Liver2017）
  • Invited
• [Presentation] B型肝炎ウイルスを標的とするproanthocyanidinの同定と作用機序の解析 (2017)
  • Author(s): 九十田千子、小嶋聡一、脇田隆字
  • Organizer: 第53回日本肝臓学会総会
• [Presentation] 1.Identification of a new class of HBV and HDV entry inhibitor, proanthocyanidin (2017)
  • Author(s): Tsukuda S, Watashi K, Kojima S, Wakita T
  • Organizer: AASLD The Liver Meeting
  • Int'l Joint Research
• [Presentation] NTCPを利用したHBV創薬研究およびNTCPを介したHBV侵入機構の解析 (2017)
  • Author(s): 九十田千子, 渡士幸一, 小嶋聡一, 脇田隆字
  • Organizer: 生命科学系学会合同年次大会 (ConBio2017)
  • Invited
• [Presentation] Characterization of origomeric flavonoids as a novel class of anti-HBV entry inhibitors that directly target viral envelope protein. (2016)
  • Author(s): Tsukuda S, Watashi K, Hojima T, Isogawa M, Iwamoto M, Suzuki R, Aizaki H, Kojima S, Sugiyama M, Saito A, Tanaka Y, Mizokami M, Wakita T
  • Organizer: 第64回日本ウイルス学術集会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2016-10-23
• [Presentation] Retinoid inhibitors abolish the host permissiveness to hepatitis B virus infection by modulating the expression of NTCP (2016)
  • Author(s): Tsukuda S, Watashi K, Iwamoto M, Suzuki R, Aizaki H, Kojima S, Sugiyama M, Saito A, Tanaka Y, Mizokami M, Wakita T
  • Organizer: 23rd International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Kyoto International Conference Center
  • Year and Date: 2016-10-11
  • Int'l Joint Research
• [Presentation] Proanthocyanidin and its analogs are new class of HBV and HDV entry inhibitors that target the viral preS1 region (2016)
  • Author(s): Tsukuda S, Watashi K, Hojima T, Isogawa M, Iwamoto M, Suzuki R, Aizaki H, Kojima S, Sugiyama M, Saito A, Tanaka Y, Mizokami M, Wakita T
  • Organizer: International meeting on molecular biology of hepatitis B viruses
  • Place of Presentation: Yonsei University
  • Year and Date: 2016-09-21
  • Int'l Joint Research
• [Presentation] B型肝炎ウイルスを標的とした新規侵入阻害剤の同定と作用機序の解析 (2016)
  • Author(s): 九十田千子, 小嶋聡一, 脇田隆字
  • Organizer: 第52回日本肝臓学会総会
  • Place of Presentation: ニューホテルオータニ幕張/東京ベイ幕張ホテル
  • Year and Date: 2016-05-19
• [Presentation] B型肝炎ウイルス粒子を直接標的とする侵入阻害剤proanthocyanidin (2016)
  • Author(s): 九十田千子、渡士幸一、濱田義知、齊藤安貴子、五十川正記、田中義正、鈴木亮介、相崎英樹、小嶋聡一、杉山真也、田中靖人、溝上雅史、脇田隆字.
  • Organizer: 第26回抗ウイルス療法学会総会
  • Place of Presentation: 名古屋東急ホテル、名古屋市立大学
  • Year and Date: 2016-05-13
• [Presentation] Inhibition of HBV entry process through targeting host and viral molecules (2016)
  • Author(s): Tsukuda S, Watashi K, Hamada Y, Isogawa M, Iwamoto M, Suzuki R, Aizaki H, Kojima S, Sugiyama M, Saito A, Tanaka Y, Mizokami M, Wakita T.
  • Organizer: 4th Taiwan-Korea-Japan Research Symposium on Hepatitis B Virus
  • Place of Presentation: Humanities and Social Sciences Building, Taiwan
  • Year and Date: 2016-04-09
  • Int'l Joint Research
• [Book] Encyclopedia of Signaling Molecules Second Edition -NTCP- (2017)
  • Author(s): Senko TsukudaMasashi IwamotoKoichi Watashi
  • Publisher: Springer, Cham
  • ISBN: 9783319671987
• [Book] Encyclopedia of Signaling Molecules, 2nd Edition (2017)
  • Author(s): Tsukuda S, Iwamoto M, Watashi K
  • Total Pages: 4700
  • Publisher: Springer Nature