デングウイルス感染蚊の網羅的遺伝子発現と新規ベクタ－コントロール

• Project/Area Number: 14J12097
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Epidemiology and preventive medicine
• Research Institution: The University of Tokyo (2015); Oita University (2014)
• Principal Investigator: ルントゥウェネ L.R. 東京大学, 新領域創成科学研究科, 特別研究員(PD)
• Project Period (FY): 2014-04-25 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥1,940,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥240,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥900,000 (Direct Cost: ¥900,000)
• Keywords: dengue virus / deep sequencing / l(2)efl / aedes aegypti / vector mosquito / RNA-seq / qRT-PCR / L(2)efl / siRNA / CCL-125 / Aedes aegypti

Outline of Annual Research Achievements

On this fiscal year, we tried to analyze the gene in-vivo by injecting l(2)efl-1, -2, -3, and -4 dsRNAs to mosquito thoraxes and then followed by DENV-2 injection 48 hours after dsRNA-injection. We used the same dsRNA that we transfected in CCL-125. We did preliminary experiment to see how long the dsRNA would give an effect in-vivo. We found that 72 hours after dsRNA injection, the level of l(2)efl-1 to l(2)efl-5 was suppressed. Because we needed to consider that after 72 hours dsRNA might be losing its effect, we decided to inject DENV-2 48 hours after dsRNA injection. Following dsRNA-injection, mosquitoes looked healthy with only little number of mosquitoes died. Forty eight hours afterwards, we injected mosquitoes with DENV-2 and sacrificed the mosquitoes 24, 48, and 72 hours later. Unfortunately, using qRT-PCR, we found that there was no significant difference between dsRNA-injected and control mosquitoes.
The reason that DENV-2 kept on replicating might because dsRNA was losing its effect at the time of DENV-2 injection. In the future, we will try to co-inject DENV-2 and dsRNA at the same time or to create a transgenic mosquito by injecting dsRNA at the embryo stage. We have submitted the results of in-vitro experiment to Japanese Journal of Infectious Disease for consideration.

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Susceptibility of Aedes flavopictus miyarai and Aedes galloisi mosquito species in Japan to dengue type 2 virus (2016)
  • Author(s): Raweewan Srisawat, Thipruethai Phanitchat, Narumon Komalamisra, Naoki Tamori, Lucky Runtuwene, Kaori Noguchi, Kyoko Hayashida, Shinya Hidano, Naganori Kamiyama, Ikuo Takashima, Tomohiko Takasaki, Ichiro Kurae, Narihiro Narita, Takashi Kobayashi, Yuki Eshita
  • Journal Title: Asian Pacific Journal of Tropical Biomedicine Volume: 6 Pages: 1-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 迫り来る蚊媒介感染症 (2015)
  • Author(s): 江下優樹,Runtuwene Lucky Ronald,林田京子,飛騨野真也,神山長慶,小林隆志
  • Journal Title: 生体の科学 Volume: 66 Pages: 352-355
  • Peer Reviewed
• [Journal Article] Vector competence of Aedes aegypti to dengue virus (2014)
  • Author(s): Lucky R. Runtuwene, Kaori Noguchi, Akinori Tokunaga, Takashi Kobayashi, Kenta Nakai, Yuki Eshita
  • Journal Title: Urban Pest Managament Volume: 4 Pages: 1-14
  • Peer Reviewed / Open Access
• [Journal Article] Dengue transmission model by means of viremic adult immuno-competent mouse (2014)
  • Author(s): Lucky Ronald Runtuwene, Eiji Konishi, Atsushi Yamanaka, Yoshihiro Makino, Yutaka Suzuki, Tomohiko Takasaki, Ichiro Kurane, Takashi Kobayashi and Yuki Eshita
  • Journal Title: Parasites & Vectors Volume: 7 Issue: 1 Pages: 143-143
  • DOI: 10.1186/1756-3305-7-143
  • Peer Reviewed / Open Access
• [Presentation] 多発生硬化症の病態形成機に関わるT細胞の活性化を制御するNOD様受容体とTRAF6の役割の解明 (2015)
  • Author(s): 神山ながのり,飛騨野真也,林田京子,Runtuwene R. Lucky,野口かおり,園田光,江下優樹,小林隆
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2015-12-01
• [Presentation] 熱帯熱マラリア原虫のシングルセルトランスクリプトーム解析 (2015)
  • Author(s): 救仁郷圭祐,Runtuwene Lucky Ronald,朝日博子,山岸潤也,鈴木穣
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2015-12-01
• [Presentation] 日本国内への侵入が危惧される節足動物媒介性ウイルスと日本の蚊について (2015)
  • Author(s): 江下優樹,福田昌子,Lucky R. Runtuwene,野口香緒里,飛弾野真也,神山長慶,林田京子,小林隆志, Thipruethai Phanitchat,Raweewan Srisawat,Narumon Komalamisra,成田弘成,牛島廣治,倉根一郎,高崎智彦
  • Organizer: 第36回都市有害生物管理学会大会・総会
  • Place of Presentation: 東京農業大学
  • Year and Date: 2015-07-04
• [Presentation] Susceptibility of Japanese Aedes (Stegomyia) mosquitoes to arboviruses (2015)
  • Author(s): Yuki Eshita, Masako Fukuda, Lucky R. Runtuwene, Kaori Noguchi, Shinya Hidano, Naganori Kamiyama, Kyoko Hayashida, Raweewan Srisawat, Narumon Komalamisra, Hironari Narita, Hiroshi Ushijima, Arthur E. Mongan, Josef Tuda, Mihoko Imada, Junya Yamagishi, Chihiro Sugimoto, Ryuichiro Maeda, Yutaka Suzuki, Takashi Kobayashi
  • Organizer: The 2nd conference on Asian Pediatric Infectious Diseases
  • Place of Presentation: The University of Tokyo
  • Year and Date: 2015-06-19
• [Presentation] 日本産Aedes (Stegomyia) scutellarisグループ蚊のチクングニアウイルス媒介能について (2015)
  • Author(s): 福田昌子、Lucky R. Runtuwene、野口香緒里、Thipruethai Phanitchat、Raweewan Srisawat、Narumon Komalamisra、牛島廣治、倉根一郎、高崎智彦、小林隆志、江下優樹
  • Organizer: 第67回日本衛生動物学会大会
  • Place of Presentation: 金沢大学宝町キャンパス（石川県金沢市）
  • Year and Date: 2015-03-27 – 2015-03-29
• [Presentation] Susceptibility of Japanese Aedes mosquitoes to arboviruses (2015)
  • Author(s): Yuki Eshita, Masako Fukuda, Lucky R. Runtuwene, Shinya Hidano, Kyoko Hayashida, Raweewan Srisawat, Narumon Komalamisra, TomohikoTakasaki, Ichiro Kurane, Hironari Narita, Hiroshi Ushijima, Junya Yamagishi, Chihiro Sugimoto, Yutaka Suzuki and Takashi Kobayashi
  • Organizer: Second International Workshop on Aedes albopictus, the dengue vector
  • Place of Presentation: Guanzou, P.R. China
  • Year and Date: 2015-03-23 – 2015-03-25
• [Presentation] RT-LAMP法の改良によるアルボウイルス症診断の検討 (2014)
  • Author(s): 岩中 愛、松原祥恵、福田昌子、Thipruethai Phanitchat、Raweewan Srisawat、Lucky R. Runtuwene、野口香緒里、Arthur E. Mongan、鈴木 穣、Narumon Komalamisra、成田弘成、森田公一、倉根一郎、 高崎智彦、林田京子、杉本千尋、小林隆志、江下優樹
  • Organizer: 第26回日本環境動物昆虫学会年次大会
  • Place of Presentation: 長崎大学大学教育学部（長崎県長崎市）
  • Year and Date: 2014-11-29 – 2014-11-30
• [Presentation] Potential novel anti-dengue in Aedes aegypti mosquito (2014)
  • Author(s): Lucky R. Runtuwene, Shuichi Kawashima, Kaori Noguchi, Yutaka Suzuki, Sumio Sugano, Kenta Nakai, Ryuichiro Maeda, Junya Yamagishi, Chihiro Sugimoto, Tomohiko Takasaki, Ichiro Kurane, Yuki Eshita and Takashi Kobayashi
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 一過性的にデングウイルス血症をマウスに起こして、定量的感染蚊を大量に作る新規の方法 (2014)
  • Author(s): 江下優樹, Lucky R.Runtuwene, 小西英二, 山中敦史, 牧野芳大,野口香緒里, 川上絵理, 福田昌子, 小林隆志, Raweewan Srisawat, Narumon Komalamisra, 成田弘成, 牛島廣治, Arthur E. Mongan, 今田美穂子,山岸潤也, 鈴木 穣, 中井謙太, 前田龍一郎, 杉本千尋, 倉根一郎, 高崎智彦
  • Organizer: 第35回都市有害生物管理学会大会・総会
  • Place of Presentation: 日本大学生産工学部（千葉県県習志野市）
  • Year and Date: 2014-06-28 – 2014-06-29
• [Book] NGSアプリケーションRNA－seq実験ハンドブック (2016)
  • Author(s): 鈴木絢子,Lucky Runtuwene,阿部佳澄
  • Publisher: 羊土社