Reveling the intra-cellular replication machinery of Severe fever with thrombocytopenia syndrome virus by using live-cell imaging.

2023 Fiscal Year Annual Research Report

• Project/Area Number: 21K07059
• Research Institution: Nagasaki University
• Principal Investigator: 高松 由基 長崎大学, 熱帯医学研究所, 准教授 (00750407)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: SFTSV / RNP形成 / RNP輸送 / NPモチーフ / 転写・複製 / ウイルス様粒子

Outline of Annual Research Achievements

本研究では、五つの到達目標を定めて、「SFTSVの細胞内複製機構を制御する新しい治療薬開発の基盤を構築する」ことを目指した。本研究では細胞内動態を評価するライブセルイメージングシステム、転写・複製を評価するミニゲノムアッセイ系、ウイルス粒子形成・出芽を評価するウイルス様粒子系を構築することで、ウイルスの分子機能解析を行う基盤を整えた。またリボヌクレオプロテイン複合体（RNP）のコアを形成するヌクレオプロテイン（NP）については、機能的ドメインの報告は限られており、RNPの形成及び他のウイルスタンパク質との相互作用に関わる領域は同定されていなかった。そこでNP変異体を構築し、各種アッセイ系で変異体の機能解析を行った。
三年間のプロジェクトで、本年は最終年であった。初年度に到達目標1：感染細胞におけるRNPおよびウイルス粒子の細胞内動態をライブセルイメージング顕微鏡で明らかにする、および到達目標2： RNPやウイルス粒子の細胞内輸送における分子機構を明らかにするための非感染性のライブセルイメージングシステムを構築する、を達成した。これにより、SFTSV感染細胞におけるウイルス粒子の動きを制御するタンパク質が表面抗原タンパク質GPであり、NPはGPと共輸送されることがわかった。2年目は、NPタンパク質のペプチドモチーフに着目し、転写・複製を制御するアミノ酸領域を同定した。さらに同定したNPペプチドモチーフについて、相互作用する宿主因子の機能解析を進めた（到達目標4：ウイルスRNP形成および輸送を介在する宿主細胞因子を同定する）。3年目は、到達目標3 ：リバースジェネティクスを用いて組換えウイルスを構築する、および、到達目標5：RNPおよびRNP様構造を精製し微細構造解析を行う、を進めた。以上を通して、ウイルスの複製機構を解明し特異的な治療薬を開発するための基盤を構築した。

Research Products

• [Journal Article] Role of pre-existing immunity in driving the dengue virus serotype 2 genotype shift in the Philippines: A retrospective analysis of serological data (2024)
  • Author(s): Balingit JC, Dimamay MPS, Suzuki R, Matsuda M, Xayavong D, Ngwe Tun MM, Matias RR, Natividad FF, Moi ML, Takamatsu Y, Culleton R, Buerano CC, Morita K.
  • Journal Title: Int J Infect Dis. Volume: 139 Pages: 59-68
  • DOI: 10.1016/j.ijid.2023.11.025
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mutations in the 3' non-coding region of a no-known vector flavivirus Yokose virus increased its replication ability in mosquito C6/36 cells (2024)
  • Author(s): Tajima S, Kataoka M, Takamatsu Y, Ebihara H, Lim CK
  • Journal Title: Virology Volume: 589:109928 Pages: 1-13
  • DOI: 10.1016/j.virol.2023
  • Peer Reviewed / Open Access
• [Journal Article] Clinical factors associated with SFTS diagnosis and severity in cats (2024)
  • Author(s): Osako Hiromu、Xu Qiang、Nabeshima Takeshi、Balingit Jean Claude、Nwe Khine Mya、Yu Fuxun、Inoue Shingo、Hayasaka Daisuke、Tun Mya Myat Ngwe、Morita Kouichi、Takamatsu Yuki
  • Journal Title: BioRxiv Volume: 2024.03.06. Pages: 1-44
  • DOI: 10.1101/2024.03.06.583784
  • Open Access / Int'l Joint Research
• [Journal Article] A Novel, Comprehensive A129 Mouse Model for Investigating Dengue Vaccines and Evaluating Pathogenesis (2023)
  • Author(s): Ngwe Tun MM, Nwe KM, Balingit JC, Takamatsu Y, Inoue S, Pandey BD, Urano T, Kohara M, Tsukiyama-Kohara K, Morita K
  • Journal Title: Vaccines (Basel) Volume: 11(12):1857 Pages: 59-68
  • DOI: 10.3390/vaccines11121857
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] An outbreak of a novel lineage of dengue virus 2 in Vietnam in 2022 (2023)
  • Author(s): Nabeshima T, Ngwe Tun MM, Thuy NTT, Hang NLK, Mai LTQ, Hasebe F, Takamatsu Y; Nagasaki University Vietnam Research Group
  • Journal Title: J Med Virol. Volume: 95(11):e29255 Pages: 1-6
  • DOI: 10.1002/jmv.29255
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Serological and Molecular Epidemiology of Chikungunya Virus Infection in Vietnam, 2017-2019 (2023)
  • Author(s): Nguyen TV, Ngwe Tun MM, Cao MT, Dao HM, Luong CQ, Huynh TKL, Nguyen TTT, Hoang TND, Morita K, Le TQM, Pham QD, Takamatsu Y, Hasebe F.
  • Journal Title: Viruses Volume: 15(10):2065 Pages: 1-19
  • DOI: 10.3390/v15102065
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clinical, Virological, and Immunological Features in Cosmopolitan Genotype DENV-2-Infected Patients during a Large Dengue Outbreak in Sri Lanka in 2017 (2023)
  • Author(s): Nwe KM, Ngwe Tun MM, Muthugala R, Nabeshima T, Balingit JC, Rajamanthri L, Jayawardana D, Attanayake S, Inoue S, Takamatsu Y, Urano T, Morita K.
  • Journal Title: Am J Trop Med Hyg. Volume: 109(4) Pages: 917-925
  • DOI: 10.4269/ajtmh.22-0780
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Burden of Chikungunya Virus Infection during an Outbreak in Myanmar (2023)
  • Author(s): Ngwe Tun MM, Kyaw AK, Nwe KM, Myaing SS, Win YT, Inoue S, Takamatsu Y, Urano T, Thu HM, Hmone SW, Thant KZ, Morita K
  • Journal Title: Viruses Volume: 15(8):1734 Pages: 1-11
  • DOI: 10.3390/v15081734
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Markerless bacterial artificial chromosome manipulation method by red proteins of phage λ mediated homologous recombination utilizing fluorescent proteins for both positive and counter selection (2023)
  • Author(s): Yoshikawa T, Misu M, Kurosu T, Takamatsu Y, Sugimoto S, Shimojima M, Ebihara H, Saijo M
  • Journal Title: Heliyon Volume: 9(8):e18983 Pages: 1-13
  • DOI: 10.1016/j.heliyon.2023.e18983
  • Peer Reviewed / Open Access
• [Journal Article] Epidemiological evidence of acute transmission of Zika virus infection in dengue suspected patients in Sri-Lanka (2023)
  • Author(s): Ngwe Tun MM, Raini SK, Fernando L, Gunawardene Y, Inoue S, Takamatsu Y, Urano T, Muthugala R, Hapugoda M, Morita K
  • Journal Title: J Infect Public Health. Volume: 16(9) Pages: 1435-1442
  • DOI: 10.1016/j.jiph.2023.07.014.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Coinfection and circulation of chikungunya virus and dengue virus in pediatric patients in Myanmar, 2019 (2023)
  • Author(s): Ngwe Tun MM, Kyaw AK, Nabeshima T, Dumre SP, Soe AM, Nwe KM, Myaing SS, Lwin EP, Win YT, Inoue S, Takamatsu Y, Urano T, Thu HM, Thant KZ, Htun ZT, Morita K
  • Journal Title: Microbes Infect. Volume: 25(6):105129 Pages: 1-9
  • DOI: 10.1016/j.micinf.2023.105129
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Clinical manifestations of dengue in relation to dengue serotype in Vung Tau province, Vietnam 2020 to 2021 (2023)
  • Author(s): Nguyen Thi Thanh Ngan, Stefania Fraenkel Calcena, Takeshi Nabeshima, Dalouny Xayavong, Jean Claude Balingit Palma, Mya Myat Ngwe Tun, Kouichi Morita, Futoshi Hasebe, and Yuki Takamatsu
  • Organizer: 第29回トガ・フラビ・ペスチウイルス研究会プログラム
• [Presentation] Development of new RT-PCR primers largely utilized in Asian dengue epidemics (2023)
  • Author(s): Stefania Fraenkel Calcena, Takeshi Nabeshima, Dalouny Xayavong, Jean Claude Balingit Palma, Mya Myat Ngwe Tun, Kouichi Morita, Futoshi Hasebe, and Yuki Takamatsu
  • Organizer: 第57回日本脳炎ウイルス生態学研究会
• [Presentation] Clinical manifestations of dengue in relation to dengue serotype in Vung Tau province, Vietnam 2020 to 2021 (2023)
  • Author(s): Nguyen Thi Thanh Ngan, Taichiro Takemura, Takeshi Nabeshima, Yuki Takamatsu, Futoshi Hasebe, etc.
  • Organizer: 第57回日本脳炎ウイルス生態学研究会
• [Presentation] Dynamics of Dengue virus in Vung Tau, Southern Vietnam (2023)
  • Author(s): Nabeshima T, Ngwe Tun MM, Thuy NTT, Hang NLK, Mai LTQ, Hasebe F, Takamatsu Y; Nagasaki University Vietnam Research Group
  • Organizer: 第70回日本ウイルス学会学術集会
  • Int'l Joint Research
• [Presentation] The important motif of nucleocapsid protein to reveal the molecular machinery of SFTSV (2023)
  • Author(s): Qiang Xu, Satoshi Taniguchi, Jean Claude Balingit, Takeshi Nabeshima, Mya Myat Ngwe Tun, Masayuki Shimojima, Yuki Takamatsu
  • Organizer: 第70回日本ウイルス学会学術集会
  • Int'l Joint Research
• [Presentation] Coinfection and circulation of chikungunya virus and dengue virus in pediatric patients in Myanmar, 2019 (2023)
  • Author(s): Mya Myat Ngwe Tun, Aung Kyaw Kyaw, Takeshi Nabeshima, Shyam Prakash Dumre, Aung Min Soe, Khine Mya Nwe, Su Su Myaing, Ei Phyu Lwin, Ye Thu Win, Shingo Inoue, Yuki Takamatsu, Takeshi Urano, Hlaing Myat Thu, Kyaw Zin Thant, Zaw Than Htun, Kouichi Morita
  • Organizer: 第70回日本ウイルス学会学術集会
  • Int'l Joint Research
• [Presentation] Insights on the mechanism of dengue virus serotype 2 genotype shift in the Philippines (2023)
  • Author(s): Jean Claude Balingit, Mark Pierre Dimamay, Ryosuke Suzuki, Mami Matsuda, Dalouny Xayavong, Mya Myat Ngwe Tun, Ronald Matias, Filipinas Natividad, Richard Culleton, Meng Ling Moi, Yuki Takamatsu, Corazon Buerano, Kouichi Morita
  • Organizer: 第70回日本ウイルス学会学術集会
  • Int'l Joint Research
• [Presentation] Characterization of cross-reactive peptides with SARS-CoV-2 VOCs in Vietnam before COVID-19 pandemic. (2023)
  • Author(s): Ngan Thi Thanh Nguyen, Linh Ngoc Nguyen, Hau Thi Bich Vu, Thuy Thi Thu Nguyen, Thanh Thi Le, Phuong Vu Mai Hoang, Hang Le Khanh Nguyen, Anh Duc Dang, Mai Thi Quynh Le, Meng Ling Moi, Yuki Takamatsu, Hasebe Futoshi, Morita Kouichi
  • Organizer: 第70回日本ウイルス学会学術集会
  • Int'l Joint Research
• [Presentation] ベトナムにおけるデングウイルス感染症研究 (2023)
  • Author(s): Yuki Takamatsu
  • Organizer: グローバルヘルス合同大会2023
  • Int'l Joint Research / Invited