Functional evaluation of the candidate proteins of hepatitis E virus receptor and identification of host factors involved in the permissibility of infection

• Project/Area Number: 19K07579
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49060:Virology-related
• Research Institution: Jichi Medical University
• Principal Investigator: Nagashima Shigeo 自治医科大学, 医学部, 講師 (60433116)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: E型肝炎ウイルス / 受容体 / 感染許容性 / 細胞内侵入 / 細胞間伝播 / 粒子形成 / 許容性 / 感染感受性 / 感染初期過程 / エクソソーム

Outline of Research at the Start

E型肝炎ウイルス(HEV)はノンエンベロープウイルスに分類されているが、培養上清中や血液中のHEV粒子は、宿主細胞のエンドソーム膜に覆われたエンベロープウイルス様の粒子形態をとる。これらの形態の異なるHEV粒子は培養細胞に対して同等の感染性を有するが、膜に覆われたHEV粒子の表面にはウイルス由来の糖蛋白質が存在していない。一方で、HEV粒子表面の膜成分はエクソソームと共通の抗原性を有することを見出した。
本研究では、これまでの研究から同定された粒子形態の異なるHEVの受容体候補蛋白質の機能評価ならびにHEVに感受性を示す細胞の許容性を規定する宿主因子を同定し、増殖過程における機能を解析する。

Outline of Final Research Achievements

Previous studies have suggested that membrane-associated hepatitis E virus (quasi-enveloped HEV : eHEV) and non-enveloped HEV (neHEV) utilize the different proteins as the receptors for infection. In this study, to evaluate the function of the receptors of eHEV and neHEV, we generated candidate receptor gene knockout cells or stable cell line that express candidate receptor proteins and analyzed virus replication using those cells. As a result, we revealed that each of the candidate receptor proteins could be a receptor for infection of eHEV or neHEV. We were also able to identify host proteins required for entry and intercellular spread of HEV.

Academic Significance and Societal Importance of the Research Achievements

HEVの感染受容体として同定された候補蛋白質の機能評価により、これらの蛋白質が、粒子形態の異なる2種類のHEVの受容体として機能していることが明らかとなった。E型肝炎に対する特異的な治療法が確立されていない現状にあって、HEVの感染初期過程に関与する宿主因子が明らかになったことで、抗ウイルス剤の開発など特異的な治療法の確立に向けた新たな研究基盤を構築できるものと考えられる。また、HEVの細胞内侵入ならびに細胞間伝播に重要な宿主蛋白質を同定することに成功した。これらの宿主蛋白質は、HEVに対する治療薬開発のための宿主側の創薬ターゲットと成り得ることが期待される。

Research Products

• [Journal Article] Identification of hepatitis E virus in wild sika deer in Japan (2022)
  • Author(s): Takahashi Masaharu、Nishizono Akira、Kawakami Manri、Fukui Emiko、Isogai Emiko、Matsuoka Hiroyuki、Yamamoto Seigo、Mizuo Hitoshi、Nagashima Shigeo、Murata Kazumoto、Okamoto Hiroaki
  • Journal Title: Virus Research Volume: 308 Pages: 198645-198645
  • DOI: 10.1016/j.virusres.2021.198645
  • Peer Reviewed
• [Journal Article] Development of Recombinant Infectious Hepatitis E Virus Harboring the nanoKAZ Gene and Its Application in Drug Screening (2022)
  • Author(s): Primadharsini Putu Prathiwi、Nagashima Shigeo、Nishiyama Takashi、Takahashi Masaharu、Murata Kazumoto、Okamoto Hiroaki
  • Journal Title: Journal of Virology Volume: 96 Issue: 6
  • DOI: 10.1128/jvi.01906-21
  • Peer Reviewed
• [Journal Article] Mechanism of Cross-Species Transmission, Adaptive Evolution and Pathogenesis of Hepatitis E Virus (2021)
  • Author(s): Primadharsini Putu Prathiwi、Nagashima Shigeo、Okamoto Hiroaki
  • Journal Title: Viruses Volume: 13 Issue: 5 Pages: 909-909
  • DOI: 10.3390/v13050909
  • Peer Reviewed / Open Access
• [Journal Article] Identification and a full genome analysis of novel camel hepatitis E virus strains obtained from Bactrian camels in Mongolia (2021)
  • Author(s): Nishizawa Tsutomu、Takahashi Masaharu、Tsatsralt-Od Bira、Nyamdavaa Khurelbaatar、Dulmaa Nyamkhuu、Osorjin Byankhuu、Tseren-Ochir Erdene-Ochir、Sharav Tumenjargal、Bayasgalan Chimedtseren、Sukhbaatar Boldbaatar、Nagashima Shigeo、Murata Kazumoto、Okamoto Hiroaki
  • Journal Title: Virus Research Volume: 299 Pages: 198355-198355
  • DOI: 10.1016/j.virusres.2021.198355
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Production of capsid proteins of rat hepatitis E virus in Escherichia coli and characterization of self-assembled virus-like particles (2021)
  • Author(s): Kobayashi Tominari、Takahashi Masaharu、Ohta Satoshi、Nagashima Shigeo、Primadharsini Putu Prathiwi、Mulyanto、Kunita Satoshi、Murata Kazumoto、Okamoto Hiroaki
  • Journal Title: Virus Research Volume: 302 Pages: 198483-198483
  • DOI: 10.1016/j.virusres.2021.198483
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Clinical and virologic features of hepatitis E virus infection at a university hospital in Japan between 2000 and 2019 (2020)
  • Author(s): Sato Naoto、Watanabe Shunji、Miura Kouichi、Morimoto Naoki、Takaoka Yoshinari、Nomoto Hiroaki、Isoda Norio、Nagashima Shigeo、Takahashi Masaharu、Okamoto Hiroaki、Yamamoto Hironori
  • Journal Title: Journal of Medical Virology Volume: 92 Issue: 12 Pages: 3572-3583
  • DOI: 10.1002/jmv.26248
  • Peer Reviewed
• [Journal Article] Prevalence and genotype/subtype distribution of hepatitis E virus (HEV) among wild boars in Japan: Identification of a genotype 5 HEV strain (2020)
  • Author(s): Takahashi Masaharu、Nishizawa Tsutomu、Sato Yukihiro、Miyazaki Shinichi、Aikawa Tatsuya、Ashida Kozo、Tamaru Tomoko、Oguro Kunihiko、Hayakawa Fumihiro、Matsuoka Hiroyuki、Ozaki Hideaki、Kodera Yuuji、Irokawa Masahiko、Hirose Hideo、Nagashima Shigeo、Kawakami Manri、Mizuo Hitoshi、Okamoto Hiroaki、Murata Kazumoto
  • Journal Title: Virus Research Volume: 287 Pages: 198106-198106
  • DOI: 10.1016/j.virusres.2020.198106
  • Peer Reviewed
• [Journal Article] A Nationwide Survey of Hepatitis E Virus Infection and Chronic Hepatitis in Heart and Kidney Transplant Recipients in Japan (2020)
  • Author(s): Owada Y, Oshiro Y, Inagaki Y, Harada H, Fujiyama N, Kawagishi N, Yagisawa T, Usui J, Akutsu N, Itabashi Y, Saito K, Watarai Y, Ichimaru N, Imamura R, Kyakuno M, Ide K, Shibuya Y, Okabe Y, Ono M, Sasaki K, Shiose A, Yamagishi K, Ohnishi H, Nagashima S, et al.
  • Journal Title: Transplantation Volume: 104 Issue: 2 Pages: 437-444
  • DOI: 10.1097/tp.0000000000002801
  • NAID: 120007132798
  • Peer Reviewed
• [Journal Article] The spontaneous clearance of hepatitis E virus (HEV) and emergence of HEV antibodies in a transfusion-transmitted chronic hepatitis E case after completion of chemotherapy for acute myeloid leukemia (2020)
  • Author(s): Okano H, Nakano T, Ito R, Tanaka A, Hoshi Y, Matsubayashi K, Asakawa H, Nose K, Tsuruga S, Tochio T, Kumazawa H, Isono Y, Tanaka H, Matsusaki S, Sase T, Saito T, Mukai K, Nishimura A, Kawakami K, Nagashima S, et al.
  • Journal Title: Clinical Journal of Gastroenterology Volume: 13 Issue: 2 Pages: 252-259
  • DOI: 10.1007/s12328-019-01024-3
  • Peer Reviewed
• [Journal Article] IFN-λ3 as a host immune response in acute hepatitis E virus infection (2020)
  • Author(s): Murata K, Kang JH, Nagashima S, Matsui T, Karino Y, Yamamoto Y, Atarashi T, Oohara M, Uebayashi M, Sakata H, Matsubayashi K, Takahashi K, Arai M, Mishiro S, Sugiyama M, Mizokami M, Okamoto H.
  • Journal Title: Cytokine Volume: 125 Pages: 154816-154816
  • DOI: 10.1016/j.cyto.2019.154816
  • Peer Reviewed
• [Journal Article] Multivesicular body sorting and the exosomal pathway are required for the release of rat hepatitis E virus from infected cells (2020)
  • Author(s): Primadharsini PP, Nagashima S, Takahashi M, Kobayashi T, Nishiyama T, Nishizawa T, Yasuda J, Mulyanto, Okamoto H.
  • Journal Title: Virus Research Volume: 278 Pages: 197868-197868
  • DOI: 10.1016/j.virusres.2020.197868
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Screening of novel drugs for inhibiting hepatitis E virus replication (2019)
  • Author(s): Nishiyama T, Kobayashi T, Jirintai S, Kii I, Nagashima S, Primadharsini PP, Nishizawa T, Okamoto H.
  • Journal Title: Journal of Virological Methods Volume: 270 Pages: 1-11
  • DOI: 10.1016/j.jviromet.2019.04.017
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Changing clinical and molecular characteristics of hepatitis E virus infection in Mie Prefecture, Japan: Disappearance of indigenous subtype 3e strains (2019)
  • Author(s): Nakano T, Okano H, Takahashi M, Nagashima S, Shiraki K, Oya Y, Inoue H, Ohmori S, Tsukimoto M, Satoshi I, Fujimoto S, Kobayashi M, Yamawaki M, Kumagai M, Ninomiya J, Maekawa T, Kojima Y, Araki J, Hamaoka S, Horiike S, Yoshimura H, Takeuchi K, Itoh K, Akachi S, Uraki S, Yamamoto N, Ogura S, Sugimoto K, et al.
  • Journal Title: Hepatology Research Volume: 49 Issue: 9 Pages: 1003-1014
  • DOI: 10.1111/hepr.13357
  • Peer Reviewed
• [Journal Article] Genetic Variability and Evolution of Hepatitis E Virus (2019)
  • Author(s): Primadharsini PP, Nagashima S, Okamoto H.
  • Journal Title: Viruses Volume: 11 Issue: 5 Pages: 456-456
  • DOI: 10.3390/v11050456
  • Peer Reviewed
• [Journal Article] Antiviral candidates against the hepatitis E virus (HEV) and their combinations inhibit HEV growth in in vitro (2019)
  • Author(s): Nishiyama T, Kobayashi T, Jirintai S, Nagashima S, Primadharsini PP, Nishizawa T, Okamoto H.
  • Journal Title: Antiviral Research Volume: 170 Pages: 104570-104570
  • DOI: 10.1016/j.antiviral.2019.104570
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Functional analysis of a highly conserved YXXΦ motif in the ORF2 protein of hepatitis E virus (2021)
  • Author(s): Shigeo Nagashima、Putu Prathiwi Primadharsini、Masaharu Takahashi、Hiroaki Okamoto、Kazumoto Murata
  • Organizer: 第68回日本ウイルス学会学術集会
• [Presentation] The tight junction protein ZO-1 is necessary for growth of hepatitis E virus in cultured cells (2019)
  • Author(s): Shigeo Nagashima, Putu Prathiwi Primadharsini, Tominari Kobayashi, Masaharu Takahashi, Hiroaki Okamoto
  • Organizer: 第67回日本ウイルス学会学術集会
• [Remarks] 自治医科大学 医学部 感染・免疫学講座 ウイルス学部門 ホームページ
  • URL: https://www.jichi.ac.jp/virology/
• [Remarks] 自治医科大学医学部 感染・免疫学講座 ウイルス学部門 ホームページ
  • URL: http://www.jichi.ac.jp/virology/