Development of novel mucosal vaccines using reovirus vectors

• Project/Area Number: 21K19379
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 49:Pathology, infection/immunology, and related fields
• Research Institution: Osaka University
• Principal Investigator: Kobayashi Takeshi 大阪大学, 微生物病研究所, 教授 (90324847)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2022: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2021: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: レオウイルス / ワクチン / ワクチンベクター

Outline of Research at the Start

哺乳類オルソレオウイルス（MRV）は、ヒトに対して安全性が高いこと、高力価のウイルス調整が容易であること、粘膜免疫誘導能を有すること等、粘膜ワクチンベクターとして優れた特性を有している。本研究では、MRV遺伝子改変技術を駆使することで、ノロウイルスおよび新型コロナウイルスを対象とした消化器および呼吸器感染症に対する粘膜ワクチンの開発基盤の確立を目的とする。

Outline of Final Research Achievements

The aim of this project is to develop mucosal vaccines against the novel coronavirus (SARS-CoV-2), which has had a significant impact on the global economy, and norovirus (NV), which causes acute gastroenteritis, using mammalian reoviruses (MRVs). We generated recombinant MRV by inserting the receptor-binding domain of the SARS-CoV-2 spike protein, fused with a 2A sequence, into the MRV S1 gene segment. For the generation of recombinant MRV expressing the NV protein, we inserted the NV VP1 gene within the MRV S gene segment. The recombinant viruses were analyzed in vitro and in vivo, providing valuable insights for the further development of MRV vectors.

Academic Significance and Societal Importance of the Research Achievements

哺乳類レオウイルス（MRV）は病原性が極めて低く、経鼻、経口投与により粘膜免疫を誘導できることから優れた粘膜ワクチンベクター候補と考えられる。本研究ではヒトノロウイルスおよび新型コロナウイルスのワクチン開発を目的として、各種ウイルス抗原を発現する組換えMRVの作製を行った。in vivoにおける解析結果から、克服すべき課題も見つかり、引き続き、開発研究を進める必要はあるものの、これらの成果は様々な呼吸器・消化器感染症に対するワクチン開発にも応用可能と考えられ、社会的意義も高い。

Research Products

• [Journal Article] Characterization of Sialic Acid-Independent Simian Rotavirus Mutants in Viral Infection and Pathogenesis (2023)
  • Author(s): Yamasaki Moeko、Kanai Yuta、Wakamura Yoshiki、Kotaki Tomohiro、Minami Shohei、Nouda Ryotaro、Nurdin Jeffery A.、Kobayashi Takeshi
  • Journal Title: Journal of Virology Volume: 97 Issue: 1
  • DOI: 10.1128/jvi.01397-22
  • Peer Reviewed
• [Journal Article] N-Glycosylation of Rotavirus NSP4 Protein Affects Viral Replication and Pathogenesis. (2023)
  • Author(s): Nurdin JA, Kotaki T, Kawagishi T, Sato S, Yamasaki M, Nouda R, Minami S, Kanai Y, Kobayashi T.
  • Journal Title: Journal of Virology Volume: 97 Issue: 1
  • DOI: 10.1128/jvi.01861-22
  • Peer Reviewed
• [Journal Article] A reverse genetics system for human rotavirus G2P[4] (2022)
  • Author(s): Hamajima Rina、Lusiany Tina、Minami Shohei、Nouda Ryotaro、Nurdin Jeffery A.、Yamasaki Moeko、Kobayashi Nobumichi、Kanai Yuta、Kobayashi Takeshi
  • Journal Title: Journal of General Virology Volume: 103 Issue: 12 Pages: 001816-001816
  • DOI: 10.1099/jgv.0.001816
  • Peer Reviewed
• [Journal Article] Development of an oncolytic mammalian orthoreovirus expressing the near-infrared fluorescent protein iRFP720 (2022)
  • Author(s): Ogawa Katsuhiro、Yamada Kentaro、Etoh Tsuyoshi、Kitagawa Masahiro、Shirasaka Yoshinori、Noguchi Kazuko、Kobayashi Takeshi、Nishizono Akira、Inomata Masafumi
  • Journal Title: Journal of Virological Methods Volume: 308 Pages: 114574-114574
  • DOI: 10.1016/j.jviromet.2022.114574
  • Peer Reviewed
• [Journal Article] Emerging norovirus GII.4 Sydney[P31] causing acute gastroenteritis outbreak in children in Japan, during COVID-19, 2021 (2022)
  • Author(s): Pham Ngan Thi Kim、Nishimura Shuichi、Shimizu-Onda Yuko、Trinh Quang Duy、Komine-Aizawa Shihoko、Khamrin Pattara、Okitsu Shoko、Sato Shintaro、Kobayashi Takeshi、Maneekarn Niwat、Hayakawa Satoshi、Ushijima Hiroshi
  • Journal Title: Journal of Infection and Chemotherapy Volume: 28 Issue: 9 Pages: 1347-1351
  • DOI: 10.1016/j.jiac.2022.05.015
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The nonstructural p17 protein of a fusogenic bat-borne reovirus regulates viral replication in virus species- and host-specific manners (2022)
  • Author(s): Nouda Ryotaro、Kawagishi Takahiro、Kanai Yuta、Shimojima Masayuki、Saijo Masayuki、Matsuura Yoshiharu、Kobayashi Takeshi
  • Journal Title: PLOS Pathogens Volume: 18 Issue: 6 Pages: e1010553-e1010553
  • DOI: 10.1371/journal.ppat.1010553
  • Peer Reviewed
• [Journal Article] Whole genome sequencing and evolutionary analysis of G8P [8] rotaviruses emerging in Japan (2022)
  • Author(s): Phan Tung、Kobayashi Masaaki、Nagasawa Koo、Hatazawa Riona、Thi Kim Pham Ngan、Miyashita Hideaki、Komoto Satoshi、Tajima Takeshi、Baba Tuneyoshi、Okitsu Shoko、Khamrin Pattara、Maneekarn Niwat、Kimura Hirokazu、Kobayashi Takeshi、Hayakawa Satoshi、Ushijima Hiroshi
  • Journal Title: VirusDisease Volume: 33 Issue: 2 Pages: 215-218
  • DOI: 10.1007/s13337-022-00765-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Whole genome sequence of an uncommon G9P[4] species A rotavirus containing DS-1-like (genotype 2) genes in Japan (2022)
  • Author(s): Phan Tung、Hatazawa Riona、Komoto Satoshi、Nishimura Shuichi、Khamrin Pattara、Pham Ngan Thi Kim、Okitsu Shoko、Kobayashi Takashi、Maneekarn Niwat、Hayakawa Satoshi、Ushijima Hiroshi
  • Journal Title: Archives of Virology Volume: 167 Issue: 7 Pages: 1603-1606
  • DOI: 10.1007/s00705-022-05462-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] An increasing trend of human sapovirus infection in Japan, 2009 to 2019: An emerging public health concern (2022)
  • Author(s): Hoque S. A., Nishimura K, Thongprachum A. , Khamrin P. ,Ngan Thi Kim Pham, Islam M. T. , Khandoker n. , Okitsu S, Onda-Shimizu Y , Dey S. K. ,Maneekarn N. , Kobayashi T, Hayakawa S, Ushijima H.
  • Journal Title: Journal of Infection and Public Health Volume: 15 Issue: 3 Pages: 315-320
  • DOI: 10.1016/j.jiph.2022.01.019
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] SARS-CoV-2 infection triggers paracrine senescence and leads to a sustained senescence-associated inflammatory response (2022)
  • Author(s): Tsuji Shunya、Minami Shohei、Hashimoto Rina、Konishi Yusuke、Suzuki Tatsuya、Kondo Tamae、Sasai Miwa、Torii Shiho、Ono Chikako、Shichinohe Shintaro、Sato Shintaro、et al.
  • Journal Title: Nature Aging Volume: 2 Issue: 2 Pages: 115-124
  • DOI: 10.1038/s43587-022-00170-7
  • Peer Reviewed / Open Access
• [Journal Article] Ultrasensitive detection of SARS-CoV-2 nucleocapsid protein using large gold nanoparticle-enhanced surface plasmon resonance (2022)
  • Author(s): Yano T, Kajisa T, Ono M, Miyasaka Y, Hasegawa Y, Saito A, Otsuka K, Sakane A, Sasaki T, Yasutomo K, Hamajima R, Kanai Y, Kobayashi T, Matsuura Y, Itonaga M, Yasui T.
  • Journal Title: Scientific Reports Volume: 12 Issue: 1 Pages: 1060-1060
  • DOI: 10.1038/s41598-022-05036-x
  • Peer Reviewed / Open Access
• [Journal Article] Species A rotavirus reverse genetics: Achievements and prospects (2021)
  • Author(s): Kobayashi T, Patton J.T., Desselberger U.
  • Journal Title: Virus Research Volume: 306 Pages: 198583-198583
  • DOI: 10.1016/j.virusres.2021.198583
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Development of an entirely plasmid-based reverse genetics system for 12-segmented double-stranded RNA viruses (2021)
  • Author(s): Nouda R, Minami S, Kanai Y, Kawagishi T, Nurdin JA, Yamasaki M, Kuwata R, Shimoda H, Maeda K, Kobayash T.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 118 Issue: 42
  • DOI: 10.1073/pnas.2105334118
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Monoreassortant Rotaviruses of Multiple G Types Are Differentially Neutralized by Sera From Infants Vaccinated With ROTARIX and RotaTeq (2021)
  • Author(s): J. R. Diller, M. H. Carter, Kanai Y, S. V. Sanchez, Kobayashi T, K. M. Ogdena.
  • Journal Title: The Journal of Infectious Diseases Volume: - Issue: 10 Pages: 1720-1729
  • DOI: 10.1093/infdis/jiab479
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The Detection of Rotavirus Antigenemia by Immunochromatographic Kits: a Case Series. (2021)
  • Author(s): Ushijima H, Hikita T, Kobayashi M, Pham NTK, Onda-Shimizu Y, Kawagishi T, Okitsu S, Kanai Y, Kobayashi T, Phan T, Hoque SA, Takanashi S, Komoto S, Kumthip K, Taniguchi K, Maneekarn N, Hayakawa S, Khamrin P.
  • Journal Title: Clinical Laboratory Volume: 67 Issue: 10/2021 Pages: 2275-2282
  • DOI: 10.7754/clin.lab.2021.210125
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] FAST Proteins: Development and Use of Reverse Genetics Systems for Reoviridae Viruses (2021)
  • Author(s): Kanai Y, Kobayashi T.
  • Journal Title: Annual Review of Virology Volume: 8 Issue: 1 Pages: 515-536
  • DOI: 10.1146/annurev-virology-091919-070225
  • Peer Reviewed
• [Journal Article] Epidemiology and genetic diversity of group A rotavirus in pediatric patients with acute gastroenteritis in Thailand, 2018-2019 (2021)
  • Author(s): Nutthawadee J, Kattareeya K, Arpaporn Y, Kanai Y, Okitsu S, Kobayashi T, Nuthapong U, Ushijima H, Niwat M, Pattara K.
  • Journal Title: Infection, Genetics and Evolution Volume: 95 Pages: 104898-104898
  • DOI: 10.1016/j.meegid.2021.104898
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] DsRNA Sequencing for RNA Virus Surveillance Using Human Clinical Samples (2021)
  • Author(s): Izum T, Morioka Y, Urayama S, Motooka D, Tamura T, Kawagishi T, Kanai Y, Kobayashi T, Ono C, Morinaga A, Tomiyama T, Iseda N, Kosai Y, Inokuchi S, Nakamura S, Tanaka T, Moriishi K, Kariwa H, Yoshizumi T, Mori M, Matsuura Y, Fukuhara T.
  • Journal Title: Viruses Volume: 13 Issue: 7 Pages: 1310-1310
  • DOI: 10.3390/v13071310
  • Peer Reviewed / Open Access
• [Journal Article] Rotavirus reverse genetics systems: Development and application (2021)
  • Author(s): Kanai Y, Kobayashi T.
  • Journal Title: Virus Research Volume: 295 Pages: 198296-198296
  • DOI: 10.1016/j.virusres.2021.198296
  • Peer Reviewed
• [Journal Article] Combining machine learning and nanopore construction creates an artificial intelligence nanopore for coronavirus detection (2021)
  • Author(s): Taniguchi M, Minami S, Ono C, Hamajima R, Morimura A, Hamaguchi S, Akeda Y, Kanai Y, Kobayashi T, Kamitani W, Terada Y, Suzuki K, Hatori N, Yamagishi Y, Washizu N, Takei H, Sakamoto O, Naono N, Tatematsu K, Washio T, Matsuura Y, Tomono K.
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 3726-3726
  • DOI: 10.1038/s41467-021-24001-2
  • Peer Reviewed / Open Access
• [Presentation] Whole Genome Characterization of HAstV5-HAstV8-HAstV1 Recombinant Strains of ORF1a-ORF1b-ORF2 Regions (2022)
  • Author(s): Pattara Khamrin, Kattareeya Kumthip, Shintaro Sato, Shoko Okitsu, Takeshi Kobayashi, Satoshi Hayakawa, Hiroshi Ushijima, Niwat Maneekarn
  • Organizer: 第68回日本ウイルス学会学術集会
• [Presentation] Recent advances in rotavirus reverse genetics research (2022)
  • Author(s): 小林剛
  • Organizer: 第68回日本ウイルス学会学術集会
  • Invited
• [Presentation] ロタウイルス感染に関与する宿主因子Tumor Associated Calcium Signal Transducer 2の機能解析 (2022)
  • Author(s): 山崎萌子、金井祐太、浜島りな、小瀧将裕、南昌平、納田遼太郎、Jeffery A. Nurdin、小林剛
  • Organizer: 第68回日本ウイルス学会学術集会
• [Presentation] ロタウイルスNSP2タンパク質の機能解析 (2022)
  • Author(s): 小瀧 将裕、金井 祐太、南 昌平、納田 遼太郎、Jeffery A. Nurdin、山﨑 萌子、小林 剛
  • Organizer: 第68回日本ウイルス学会学術集会
• [Presentation] マウス順化サルロタウイルスの作出 (2022)
  • Author(s): 金井 祐太、小瀧 将裕、南 昌平、納田 遼太郎、山﨑 萌子、小林 剛
  • Organizer: 第68回日本ウイルス学会学術集会
• [Presentation] ヒトロタウイルスHN126株におけるリバースジェネティクス系の確立 (2022)
  • Author(s): 浜島 りな、Tina Lusiany、南 昌平、納田 遼太郎、Jeffery A. Nurdin、山﨑 萌子、小林 宣道、金井 祐太、小林 剛
  • Organizer: 第68回日本ウイルス学会学術集会
• [Presentation] Single amino acid substitution in VP7 promotes the pathogenicity of simian rotavirus in mice (2022)
  • Author(s): Yuta Kanai, Eriko Hirai, Tomohiro Kotaki and Takeshi Kobayashi
  • Organizer: 14th International dsRNA Virus Symposium
  • Int'l Joint Research
• [Presentation] N-glycosylation of rotavirus NSP4 protein affects viral replication and pathogenesis (2022)
  • Author(s): Tomohiro Kotaki, Jeffery A. Nurdin, Takahiro Kawagishi, Shintaro Sato, Moeko Yamasaki, Ryotaro Nouda, Shohei Minami, Yuta Kanai, Takeshi Kobayashi
  • Organizer: 14th International dsRNA Virus Symposium
  • Int'l Joint Research
• [Presentation] Analysis of the role of rotavirus NSP4 protein in viral replication and pathogenicity (2022)
  • Author(s): Tomohiro Kotaki, Nurdin Jeffery A., Kawagishi Takahiro, Sato Shintaro, Yamasaki Moeko, Nouda Ryotaro, Minami Shohei, Kanai Yuta, Kobayashi Takeshi
  • Organizer: 第20回 あわじ感染と免疫国際フォーラム
  • Int'l Joint Research
• [Presentation] Identification of TACSTD2 as a host factor involved in rotavirus infection (2022)
  • Author(s): Moeko Yamasaki, Yuta Kanai, Rina Hamajima, Tomohiro Kotaki, Shohei Minami, Ryotaro Nouda, Takeshi Kobayashi.
  • Organizer: 第20回 あわじ感染と免疫国際フォーラム
  • Int'l Joint Research
• [Presentation] A genome-wide CRISPR-Cas9 screening approach to identify host factors for rotavirus infection CRISPR-Cas9法によるロタウイルス感染に関与する宿主因子のゲノムワイドスクリーニング (2021)
  • Author(s): 山﨑 萌子、浜島 りな、金井 祐太、小瀧 将裕、南 昌平、納田 遼太郎、Nurdin Jeffery、小林 剛
  • Organizer: 第68回ウイルス学会学術集会
• [Presentation] Altered infectivity of rotaviruses in mice treated with antibiotics 抗生物質を投与したマウスにおけるロタウイルスの増殖能 (2021)
  • Author(s): 金井 祐太、Jeffery Nurdin、南 昌平、納田 遼太郎、山崎 萌子、小林 剛
  • Organizer: 第68回ウイルス学会学術集会
• [Presentation] Development of an entirely plasmid-based reverse genetics system for 12-segmented double-stranded RNA virus. (2021)
  • Author(s): Ryotaro Nouda, Shohei Minami, Yuta Kanai, Takahiro Kawagishi, Jeffery A. Nurdin, Moeko Yamasaki, Ryusei Kuwata, Hiroshi Shimoda, Ken Maeda, Takeshi Kobayashi.
  • Organizer: 第68回ウイルス学会学術集会
• [Presentation] Determining the role of rotavirus NSP4 N-glycosylation sites using reverse genetics approach (2021)
  • Author(s): Jeffery A. Nurdin, Yuta Kanai, Tomohiro Kotaki, Takahiro Kawagishi, Shohei Minami, Ryotaro Nouda, Moeko Yamasaki, Takeshi Kobayashi.
  • Organizer: 第68回ウイルス学会学術集会
• [Presentation] Analysis of intestinal epithelial cells differentiated from human induced pluripotent stem cells as a more suitable tool for the study of SARS-CoV-2 (2021)
  • Author(s): Shohei Minami, Naomi Matsumoto, Ryotaro Nouda, Jeffery Nurdin, Moeko Yamasaki, Tomohiro Kotaki, Yuta Kanai, Shintaro Sato, Takeshi Kobayashi.
  • Organizer: 第68回ウイルス学会学術集会
• [Book] 医学のあゆみ280巻9号 ウイルスを創る，ウイルスを視る (2021)
  • Author(s): 金井祐太、納田遼太郎、小林 剛
  • Publisher: 医歯薬出版株式会社
• [Book] 創薬研究者がこれだけは知っておきたい最新のウイルス学 (2021)
  • Author(s): 金井祐太、納田遼太郎、山﨑萌子、小林 剛
  • Publisher: 技術情報協会
  • ISBN: 9784861048555