Study of human norovirus receptor for developing of susceptible cell line

• Project/Area Number: 15H04738
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: Kitasato University (2016-2018); National Institute of Infectious Diseases (2015)
• Principal Investigator: KATAYAMA KAZUHIKO 北里大学, 感染制御科学府, 教授 (60342903)
• Co-Investigator(Kenkyū-buntansha): 中西 章 国立研究開発法人国立長寿医療研究センター, 老化制御研究部, 室長 (60397049)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2018: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2017: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2016: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: ノロウイルス / レセプター / CRISPR/Cas9 / ホモログ / MNV / HuNoV / CD300lf, ld / ヒト腸管オルガノイド / HuNoV感受生細胞mRNAライブラリー / トランスクリプトームサブトラクション / ヒトノロウイルス / 感受性細胞 / 増殖培養系 / ネズミノロウイルス / CD300lf, CD300ld / ヒトノロウイルス感受性細胞 / mRNAトランスクリプトーム解析 / 感受生細胞 / ノロウイルスレセプター / ノロウイルス感受生細胞 / ノロウイルス感染モデル動物 / ヒト・マウスキメラノロウイルス / オルガノイド

Outline of Final Research Achievements

Human norovirus (HNV) is the leading cause of acute gastroenteritis worldwide. Since the discovery of HNV, an efficient and reproducible norovirus replication system has not been established in cultured cells. Those results imply that the identification of a functional cell-surface receptor for norovirus might be the key to establishing a norovirus culture system. Using a genome-wide CRISPR/Cas9 guide RNA library, we identified murine CD300lf and CD300ld as functional receptors for murine norovirus (MNV). The treatment of susceptible cells with polyclonal antibody against CD300lf significantly reduced the production of viral progeny. Furthermore, CD300ld, which has an amino acid sequence in the N-terminal region of its extracellular domain that is highly homologous to that of CD300lf, also functions as a receptor for MNV.

Academic Significance and Societal Importance of the Research Achievements

MNVのレセプター分子CD300lfとCD300ldを発見した。CD300lfとCD300ldのヒトホモログはHNVのレセプターとして機能しなかった。HNVは 組織血液型抗原、MNVはシアル酸など、糖鎖がレセプターだと考えられていた。しかし、本研究でMNVレセプターが膜タンパク質であることが明らかになったことで、HNVレセプターが膜タンパク質である可能性が急浮上することになった。さらに、ヒト腸管上皮の構造を試験管内で再現可能なオルガノイドの平板培養技術の開発に成功したことにより、HNVの感染感受生細胞を再現性良く作出することが可能となり、HNVレセプター探求に新たな道を開いた。

Research Products

• [Journal Article] ノロウイルス総説 (2019)
  • Author(s): 片山和彦
  • Journal Title: 感染制御と予防衛生 Volume: 3 Pages: 4-11
  • Peer Reviewed
• [Journal Article] A Genetic Determinant of Norovirus for Susceptibility to Free Chlorine (2018)
  • Author(s): Andri Rachmadi, Masaaki Kitajima, Kozo Watanabe, Sakiko Yaegashi, Joeselle Serrana, Arata Nakamura, Toyoko Nakagomi, Osamu Nakagomi, Kazuhiko Kazuhiko Katayama, Satoshi Okabe, and Daisuke Sano
  • Journal Title: Applied and Environmental Microbiology Volume: 印刷中 Issue: 13
  • DOI: 10.1128/aem.00244-18
  • Peer Reviewed / Open Access
• [Journal Article] 院内・施設内感染症に関わる胃腸炎ウイルス (2018)
  • Author(s): 片山和彦
  • Journal Title: 感染制御と予防衛生 Volume: 2 Pages: 66-72
  • Peer Reviewed
• [Journal Article] Viral population changes during murine norovirus propagation in RAW 264.7 cells. (2017)
  • Author(s): 7.Kitamoto T, Takai-Todaka R, Kato A, Kanamori K, Takagi H, Yoshida K, Katayama K, Nakanishi A.
  • Journal Title: Front Microbiol. Volume: 8 Pages: 1091-1091
  • Peer Reviewed
• [Journal Article] Effects of disinfectants against norovirus virus-like particles predict norovirus inactivation. (2016)
  • Author(s): Sato J, Miki M, Kubota H, Hitomi J, Tokuda H, Takai-Todaka R and Katayama K.
  • Journal Title: Microbiol Immunol Volume: 60 Pages: 609-616
  • NAID: 40020945581
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Complete Genome Sequence of a Recombinant GII.P16-GII.4 Norovirus Detected in Kawasaki City, Japan, 2016. (2016)
  • Author(s): Matsushima Y, Shimizu T, Ishikawa M, Komane A, Okabe N, Ryo A, Kimura H, Katayama K, and Shimizu H.
  • Journal Title: Genome Announcements Volume: 4
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Functional receptor molecules CD300lf and CD300ld enable murine norovirus to internalize into host cells. (2016)
  • Author(s): Haga K, Fujimoto A, Takai-Todaka R, Miki M, Doan Y. H, Murakami K, Yokoyama M, Murata M, Nakanishi A, and Katayama K.
  • Journal Title: Proc Natl Acad Sci U S A Volume: 113(41)
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Molecular evolution of the capsid gene in human norovirus genogroup II. (2016)
  • Author(s): Kobayashi M, Matsushima Y, Motoya T, Sakon N, Shigemoto N, Okamoto-Nakagawa R, Nishimura K, Yamashita Y, Kuroda M, Saruki N, Ryo A, Saraya T, Morita Y, Shirabe K, Ishikawa M, Takahashi T, Shinomiya H, Okabe N, Nawasawa K, Suzuki Y, Katayama K, Kimura H.
  • Journal Title: Sci Rep. Volume: 7,6 Issue: 1 Pages: 29400-29400
  • DOI: 10.1038/srep29400
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Examination of a plasmid-based reverse genetics system for human astrovirus. (2015)
  • Author(s): Chapellier B, Tange S, Tasaki H, Yoshida K, Zhou Y, Sakon N, Katayama K, Nakanishi A.
  • Journal Title: Microbiol Immunol. Volume: 59 Issue: 10 Pages: 586-596
  • DOI: 10.1111/1348-0421.12317
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Molecular Evolution of the Capsid Gene in Norovirus Genogroup I. (2015)
  • Author(s): Kobayashi M, Yoshizumi S, Kogawa S, Takahashi T, Ueki Y, Shinohara M, Mizukoshi F, Tsukagoshi H, Sasaki Y, Suzuki R, Shimizu H, Iwakiri A, Okabe N, Shirabe K, Shinomiya H, Kozawa K, Kusunoki H, Ryo A, Kuroda M, Katayama K, Kimura H.
  • Journal Title: Sci Rep. Volume: 5 Issue: 1 Pages: 13806-13806
  • DOI: 10.1038/srep13806
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Study of norovirus infection mechanism (2018)
  • Author(s): Kazuhiko Katayama, Reiko Takai-Todaka, Chihong Song, Motohiro Miki, Akira Fujimoto, Masaru Yokoyama, Naoyuki Miyazaki, Kenji Iwasaki, and Kazuyoshi Murata.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Selective pressure of free chlorine disinfection on murine norovirus (2018)
  • Author(s): Andri Taruna Rachmadi，Masaaki Kitajima，Kozo Watanabe，Sakiko Yaegashi， Joeselle Serrana，Arata Nakamura，Toyoko Nakagomi，Osamu Nakagomi， Kazuhiko Katayama，Satoshi Okabe，Daisuke Sano.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Possible alternative: Mechanism of bile-requiring GII.3 human norovirus replication in human intestinal enteroids (2018)
  • Author(s): Kosuke Murakami, Victoria R. Tenge, Umesh Karandikar, Sasirekha Ramani, Khalil Ettayebi, Xi-Lei Zeng, Sue E. Crawford, Kazuhiko Katayama, Robert L. Atmar, Mary K. Estes.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Capsid structure of murine norovirus revealed by cry-electron microscopy (2018)
  • Author(s): Song C, Todaka R, Haga K, Fujimoto A, Yokoyama M, Miyazaki N, Iwasaki K, Katayama K, Murata K.
  • Organizer: 生物物理学会
• [Presentation] Study of norovirus virion structure and new antigen delivery system (2018)
  • Author(s): Katayama K., Takai-Todaka R., Song C., Miki M., Fujimoto A., Yokoyama M., Miyazaki N., Iwasaki K., Murata K.
  • Organizer: U.S.-Japan Cooperative Medical Sciences Program (USJCMSP) 20th International Conference on Emerging Infectious Diseases in the Pacific Rim
  • Int'l Joint Research
• [Presentation] Study of norovirus infection mechanism (2018)
  • Author(s): Katayama K., Takai-Todaka R., Song C., Miki M., Fujimoto A., Yokoyama M., Miyazaki N., Iwasaki K., Murata K.
  • Organizer: U.S.-Japan Cooperative Medical Sciences Program (USJCMSP) 20th International Conference on Emerging Infectious Diseases in the Pacific Rim
  • Int'l Joint Research
• [Presentation] Towards development of a human norovirus culture system (2018)
  • Author(s): Fujimoto A., Takai-Todaka R., Sugimoto S., Matano M., Sato T., Katayama K.
  • Organizer: U.S.-Japan Cooperative Medical Sciences Program (USJCMSP) 20th International Conference on Emerging Infectious Diseases in the Pacific Rim
  • Int'l Joint Research
• [Presentation] ノロウイルス研究の最前線 (2017)
  • Author(s): 片山和彦
  • Organizer: 日本獣医師学会
  • Place of Presentation: 金沢
  • Year and Date: 2017-02-24
  • Invited
• [Presentation] Identification of the functional receptor for murine noro virus (2017)
  • Author(s): Kazuhiko Katayama
  • Organizer: The 2017 Japan-NIH Joint symposium
  • Place of Presentation: 仙台
  • Year and Date: 2017-02-15
  • Int'l Joint Research / Invited
• [Presentation] Cultivation of human norovirus using human duodenal enteroids. (2016)
  • Author(s): Akira Fujimoto, Kei Haga, Shinya Sugimoto, Toshiro Sato, Yen Hai Doan, Motohiro Miki, Reiko Takai-Todaka, and Kazuhiko Katayama.
  • Organizer: 日本ウイルス学会
  • Place of Presentation: 札幌
  • Year and Date: 2016-10-23
• [Presentation] Identification of the functional receptor for murine norovirus (2016)
  • Author(s): Kei Haga, Akira Fujimoto, Yen Hai Doan, Reiko Takai-Todaka, Motohiro Miki, Kosuke Murakami, Masaru Youkoma, Kazuyoshi Murata, Akira Nakanishi and Kazuhiko Katayama.
  • Organizer: 日本ウイルス学会
  • Place of Presentation: 札幌
  • Year and Date: 2016-10-23
• [Presentation] Cultivation of human norovirus using human duodenal organoids. (2016)
  • Author(s): Akira Fujimoto, Kei Haga, Shinya Sugimoto, Toshiro Sato, Yen Hai Doan, Motohiro Miki, Reiko Takai-Todaka, and Kazuhiko Katayama.
  • Organizer: 6th international Calicivirus Conference
  • Place of Presentation: Savannah, GA, USA
  • Year and Date: 2016-10-09
  • Int'l Joint Research
• [Presentation] Identification of the functional receptor for murine norovirus (2016)
  • Author(s): Kei Haga, Akira Fujimoto, Yen Hai Doan, Reiko Takai-Todaka, Motohiro Miki, Kosuke Murakami, Masaru Youkoma, Kazuyoshi Murata, Akira Nakanishi and Kazuhiko Katayama.
  • Organizer: 6th international Calicivirus Conference
  • Place of Presentation: Savannah, GA, USA
  • Year and Date: 2016-10-09
  • Int'l Joint Research
• [Presentation] Norovirus Special Workshop (2016)
  • Author(s): Kazuhiko Katayama
  • Organizer: 5th International Society for Food and Environmental Virology
  • Place of Presentation: 草津
  • Year and Date: 2016-09-13
  • Int'l Joint Research / Invited
• [Presentation] Development of norovirus in cell replication efficiency monitoring system based on reverse genetics system (2015)
  • Author(s): 戸高玲子、村上耕介、朴英斌、吉田和央、中西章、片山和彦
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] Functional Analysis of Human Norovirus RNA-dependent RNA Polymerase (2015)
  • Author(s): 朴英斌、戸高玲子、芳賀慧、下池貴志、福士秀悦、染谷雄一、横山勝、佐藤裕徳、河合文啓、朴三用、片山和彦
  • Organizer: 日本ウイルス学会
  • Place of Presentation: 福岡
  • Year and Date: 2015-11-22
• [Book] 決定版 オルガノイド実験スタンダード (2019)
  • Author(s): 佐藤 俊朗、武部 貴則、永樂 元次
  • Total Pages: 368
  • Publisher: 羊土社
  • ISBN: 9784758122399
• [Book] ワクチン (2018)
  • Author(s): 日本ワクチン学会
  • Total Pages: 376
  • Publisher: 朝倉書店
  • ISBN: 9784254301151
• [Book] ウイルス検査法 (2018)
  • Author(s): 日本臨床ウイルス学会
  • Total Pages: 382
  • Publisher: 春恒社
  • ISBN: 9784915698132
• [Patent(Industrial Property Rights)] ヒト下痢症ウイルスの感染・増殖培養用２Ｄオルガノイド及びその使用 (2017)
  • Inventor(s): 佐藤俊朗、片山和彦 他
  • Industrial Property Rights Holder: 佐藤俊朗、片山和彦 他
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-163711
  • Filing Date: 2017
  • Overseas
• [Patent(Industrial Property Rights)] ノロウイルスが増殖可能な遺伝子組換え培養細胞、及び、その用途 (2016)
  • Inventor(s): 平成28年2月2日
  • Industrial Property Rights Holder: 平成28年2月2日
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-02-02