Identification and functional analysis of host factors involved in early steps of hepatitis E virus infection

• Project/Area Number: 16K08817
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Virology
• Research Institution: Jichi Medical University
• Principal Investigator: Nagashima Shigeo 自治医科大学, 医学部, 講師 (60433116)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: E型肝炎ウイルス / 感染初期過程 / 受容体 / 細胞内侵入 / 感染感受性 / エクソソーム / エンベロープ / 細胞侵入 / マイクロアレイ / 細胞内輸送

Outline of Final Research Achievements

In this study, to identify the possible entry receptors for hepatitis E virus (HEV) infection, we investigated the antigenicity and morphological features of the membrane-associated HEV particles. Our results indicated that membrane components are common between the membrane-associated HEV particles and exosomes, and that the capsids of HEV particles are individually covered by lipid membranes. In addition, our results suggested that the two candidate proteins identified in our previous study could serve as different entry receptors for two distinct forms of HEV particles.

Academic Significance and Societal Importance of the Research Achievements

これまでの解析からHEVの感染受容体としての関与が示唆されている候補蛋白質に対して、siRNAを用いたスクリーニングを行った。その結果、どちらか一方の粒子形態でのみ増殖効率が低下する蛋白質を同定することができた。また、感染阻害実験やウイルスとの結合実験により、これらの蛋白質がHEVの感染受容体として機能することを示すことができた。感染受容体はHEVの感染初期過程に非常に重要な分子であるため、両者の結合を阻害するような化合物を探索することで、治療薬開発への基礎的知見を提供できるものと期待される。

Research Products

• [Journal Article] An analysis of two open reading frames (ORF3 and ORF4) of rat hepatitis E virus genome using its infectious cDNA clones with mutations in ORF3 or ORF4 (2018)
  • Author(s): Tanggis、Kobayashi Tominari、Takahashi Masaharu、Jirintai Suljid、Nishizawa Tsutomu、Nagashima Shigeo、Nishiyama Takashi、Kunita Satoshi、Hayama Emiko、Tanaka Takeshi、Mulyanto、Okamoto Hiroaki
  • Journal Title: Virus Research Volume: 249 Pages: 16-30
  • DOI: 10.1016/j.virusres.2018.02.014
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hepatitis E virus subtype 3f strains isolated from Japanese hepatitis patients with no history of travel to endemic areas ? The origin analyzed by molecular evolution (2018)
  • Author(s): Nakano Tatsunori、Takahashi Masaharu、Takahashi Kazuaki、Nagashima Shigeo、Suzuki Yusuke、Nishigaki Yoichi、Tomita Eiichi、Okano Hiroshi、Oya Yumi、Shiraki Katsuya、Takase Kojiro、Sugimoto Kazushi、Koyama Junichi、Mizuo Hitoshi、Ikezawa Kazuto、Aikawa Tatsuya、Arai Masahiro、Okamoto Hiroaki
  • Journal Title: Virology Volume: 513 Pages: 146-152
  • DOI: 10.1016/j.virol.2017.08.008
  • Peer Reviewed
• [Journal Article] Full-length genomic sequence analysis of new subtype 3k hepatitis E virus isolates with 99.97% nucleotide identity obtained from two consecutive acute hepatitis patients in a city in northeast Japan (2017)
  • Author(s): Miura Masahito、Inoue Jun、Tsuruoka Mio、Nishizawa Tsutomu、Nagashima Shigeo、Takahashi Masaharu、Shimosegawa Tooru、Okamoto Hiroaki
  • Journal Title: Journal of Medical Virology Volume: 89 Issue: 6 Pages: 1116-1120
  • DOI: 10.1002/jmv.24743
  • Peer Reviewed
• [Journal Article] Autochthonous sporadic acute hepatitis E caused by two distinct subgenotype 3b hepatitis E virus strains with only 90% nucleotide identity (2017)
  • Author(s): Yamaguchi Yasuko、Takagi Hitoshi、Suzuki Yuhei、Maruhashi Kyoko、Kosone Takashi、Kakizaki Satoru、Sato Ken、Yamada Masanobu、Nagashima Shigeo、Takahashi Masaharu、Okamoto Hiroaki
  • Journal Title: Clinical Journal of Gastroenterology Volume: 10 Issue: 2 Pages: 168-173
  • DOI: 10.1007/s12328-017-0718-3
  • Peer Reviewed
• [Journal Article] Distinct changing profiles of hepatitis A and E virus infection among patients with acute hepatitis in Mongolia: The first report of the full genome sequence of a novel genotype 1 hepatitis E virus strain (2017)
  • Author(s): Tsatsralt-Od Bira、Primadharsini Putu Prathiwi、Nishizawa Tsutomu、Ohnishi Hiroshi、Nagashima Shigeo、Takahashi Masaharu、Jirintai Suljid、Nyamkhuu Dulmaa、Okamoto Hiroaki
  • Journal Title: Journal of Medical Virology Volume: 90 Issue: 1 Pages: 84-92
  • DOI: 10.1002/jmv.24907
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Full-length genome of a novel genotype 3 hepatitis E virus strain obtained from domestic pigs in Japan (2017)
  • Author(s): Primadharsini PP, Miyake M, Kunita S, Nishizawa T, Takahashi M, Nagashima S, Tanggis, Ohnishi H, Kobayashi T, Nishiyama T, Jirintai S, Okamoto H
  • Journal Title: Virus Research Volume: 240 Pages: 147-153
  • DOI: 10.1016/j.virusres.2017.08.003
  • Peer Reviewed
• [Journal Article] Characterization of the Quasi-Enveloped Hepatitis E Virus Particles Released by the Cellular Exosomal Pathway (2017)
  • Author(s): Nagashima Shigeo、Takahashi Masaharu、Kobayashi Tominari、Nishizawa Tsutomu、Nishiyama Takashi、Primadharsini Putu Prathiwi、Okamoto Hiroaki、Tanggis
  • Journal Title: Journal of Virology Volume: 91 Issue: 22
  • DOI: 10.1128/jvi.00822-17
  • Peer Reviewed
• [Journal Article] Full-length genomic sequences of new subtype 1g hepatitis E virus strains obtained from four patients with imported or autochthonous acute hepatitis E in Japan (2017)
  • Author(s): Nishizawa Tsutomu、Primadharsini Putu Prathiwi、Namikawa Masashi、Yamazaki Yuichi、Uraki Satoko、Okano Hiroshi、Horiike Shinichiro、Nakano Tatsunori、Takaki Shintaro、Kawakami Manri、Nagashima Shigeo、Takahashi Masaharu、Okamoto Hiroaki
  • Journal Title: Infection, Genetics and Evolution Volume: 55 Pages: 343-349
  • DOI: 10.1016/j.meegid.2017.10.007
  • Peer Reviewed
• [Journal Article] Clinical and virological features of acute hepatitis E in Gunma prefecture, Japan between 2004 and 2015. (2017)
  • Author(s): Yamazaki Y, Naganuma A, Arai Y, Takeuchi S, Kobayashi T, Takakusagi S, Hatanaka T, Hoshino T, Namikawa M, Hashizume H, Takizawa D, Ohyama T, Suzuki H, Horiguchi N, Takagi H, Sato K, Kakizaki S, Kusano M, Nagashima S, Takahashi M, Okamoto H, Yamada M.
  • Journal Title: Hepatology Research Volume: 47 Issue: 5 Pages: 435-445
  • DOI: 10.1111/hepr.12765
  • Peer Reviewed
• [Journal Article] Characterization and epitope mapping of monoclonal antibodies raised against rat hepatitis E virus capsid protein: An evaluation of their neutralizing activity in a cell culture system. (2016)
  • Author(s): Kobayashi T, Takahashi M, Tanggis, Mulyanto, Jirintai S, Nagashima S, Nishizawa T, Okamoto H.
  • Journal Title: Journal of Virological Methods Volume: 233 Pages: 78-88
  • DOI: 10.1016/j.jviromet.2016.03.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Analysis of adaptive mutations selected during the consecutive passages of hepatitis E virus produced from an infectious cDNA clone. (2016)
  • Author(s): Nagashima S, Kobayashi T, Tanaka T, Tanggis, Jirintai S, Takahashi M, Nishizawa T, Okamoto H.
  • Journal Title: Virus Research Volume: 223 Pages: 170-180
  • DOI: 10.1016/j.virusres.2016.07.011
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Production of monoclonal antibodies against the ORF3 protein of rat hepatitis E virus (HEV) and demonstration of the incorporation of the ORF3 protein into enveloped rat HEV particles. (2016)
  • Author(s): Takahashi M, Kobayashi T, Tanggis, Jirintai S, Mulyanto, Nagashima S, Nishizawa T, Kunita S, Okamoto H.
  • Journal Title: Archives of Virology Volume: 161 Pages: 3391-3404
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Cathepsin L is required for entry step of hepatitis E virus (2018)
  • Author(s): Shigeo Nagashima, Putu Prathiwi Primadharsini, Tominari Kobayashi, Masaharu Takahashi, Takashi Nishiyama, Tsutomu Nishizawa, Hiroaki Okamoto
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Cathepsin L is required for entry step of hepatitis E virus (2017)
  • Author(s): Shigeo Nagashima, Masaharu Takahashi, Tominari Kobayashi, Tanggis, Tsutomu Nishizawa, Takashi Nishiyama, Putu Prathiwi Primadharsini, Hiroaki Okamoto
  • Organizer: International Union of Microbiological Societies 2017 Singapore
  • Int'l Joint Research
• [Presentation] Capsids of hepatitis E virus particles are individually covered by lipid membranes (2017)
  • Author(s): Shigeo Nagashima, Masaharu Takahashi, Tominari Kobayashi, Tanggis, Putu Prathiwi Primadharsini, Takashi Nishiyama, Tsutomu Nishizawa, Hiroaki Okamoto
  • Organizer: 第65回日本ウイルス学会学術集会 大阪
• [Presentation] Characterization of hepatitis E virus cell entry (2016)
  • Author(s): Shigeo Nagashima, Tominari Kobayashi, Tanggis, Masaharu Takahashi, Tsutomu Nishizawa, Hiroaki Okamoto
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2016-10-23
• [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/