Mechanistic analysis of viral replication using anti-DENV compound

• Project/Area Number: 15K19109
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Virology
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: HISHIKI Takayuki 公益財団法人東京都医学総合研究所, ゲノム医科学研究分野, 主任研究員 (30535761)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: デングウイルス / 複製機構 / 抗ウイルス化合物 / ケミカルバイオロジー / 抗ウイルス剤 / 抗ウイルス

Outline of Final Research Achievements

Dengue virus (DENV) infectious disease is a major public health problem worldwide; however, specific antiviral drugs against this infection are not available. To identify novel anti-DENV compounds, we screened pharmacologically active compounds using focus reduction assay. We identified some compounds that have potent anti-DENV activity and low cytotoxicity. Time-of-drug-addition and time-of-drug-elimination assays revealed that compound inhibits translation and/or replication steps in the DENV life cycle. A subgenomic replicon system was used to verify that compound restricts RNA replication step. Furthermore, analysis of compound-resistant clone suggested that compound targets viral nonstructural protein.

Research Products

• [Journal Article] Evaluation of Macaca radiata as a non-human primate model of Dengue virus infection. (2018)
  • Author(s): Kato Fumihiro, Ishida Yuki, Kawakami Akihiko, Takasaki Tomohiko, Saijo Masayuki, Miura Tomoyuki, Hishiki Takayuki.
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 3421-3421
  • DOI: 10.1038/s41598-018-21582-9
  • NAID: 120006523833
  • Peer Reviewed / Open Access
• [Journal Article] Hirsutine, an Indole Alkaloid of Uncaria rhynchophylla, Inhibits Late Step in Dengue Virus Lifecycle (2017)
  • Author(s): Hishiki Takayuki、Kato Fumihiro、Tajima Shigeru、Toume Kazufumi、Umezaki Masahito、Takasaki Tomohiko、Miura Tomoyuki
  • Journal Title: Frontiers in Microbiology Volume: 8 Pages: 1674-1674
  • DOI: 10.3389/fmicb.2017.01674
  • Peer Reviewed / Open Access
• [Journal Article] Novel antiviral activity of bromocriptine against dengue virus replication. (2016)
  • Author(s): Kato F, Ishida Y, Oishi S, Fujii N, Watanabe S, Vasudevan SG, Tajima S, Takasaki T, Suzuki Y, Ichiyama K, Yamamoto N, Yoshii K, Takashima I, Kobayashi T, Miura T, Igarashi T, Hishiki T
  • Journal Title: Antiviral Res. Volume: 131 Pages: 141-147
  • DOI: 10.1016/j.antiviral.2016.04.014
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Dengue Virus Reporter Replicon is a Valuable Tool for Antiviral Drug Discovery and Analysis of Virus Replication Mechanisms. (2016)
  • Author(s): Kato Fumihiro, and Hishiki Takayuki.
  • Journal Title: Viruses. Volume: 8 Issue: 5 Pages: 122-122
  • DOI: 10.3390/v8050122
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Generation of a neutralization-resistant CCR5 tropic SHIV-MK38 molecular clone, a derivative of SHIV-89.6. (2016)
  • Author(s): Ishida Yuki, Yoneda Mai, Otsuki Hiroyuki, Watanabe Yuji, Kato Fumihiro, Matuura Kanako, Kikukawa Minako, Matsushita Shuzo, Hishiki Takayuki, Igarashi Tatsuhiko, Miura Tomoyuki.
  • Journal Title: J Gen Virol Volume: 97(5) Issue: 5 Pages: 1249-60
  • DOI: 10.1099/jgv.0.000421
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of RyDEN (C19orf66) as an Interferon-Stimulated Cellular Inhibitor against Dengue Virus Replication. (2016)
  • Author(s): Suzuki Youichi, Chin Weixin, Han Qien, Ichiyama Koji, Lee Ching, Eyo Zhin, Ebina Hirotaka, Takahashi Hirotaka, Takahashi Chikako, Tan Beng, Hishiki Takayuki, Ohba Kenji, Matsuyama Toshifumi, Koyanagi Yoshio, Tan Yee, Sawasaki Tatsuya, Chu Justin, Vasudevan Subhash, Sano Kouichi, Yamamoto Naoki.
  • Journal Title: PLoS Pathog Volume: 6;12(1) Issue: 1 Pages: e1005357-e1005357
  • DOI: 10.1371/journal.ppat.1005357
  • NAID: 120005763386
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hepatitis C virus utilizes VLDLR as a novel entry pathway. (2016)
  • Author(s): Ujino Saneyuki, Nishitsuji Hironori, Hishiki Takayuki, Sugiyama Kazuo, Takaku Hiroshi, Shimotohno Kunitada.
  • Journal Title: Proc Natl Acad Sci USA Volume: 5;113(1) Issue: 1 Pages: 188-93
  • DOI: 10.1073/pnas.1506524113
  • Peer Reviewed
• [Journal Article] Apolipoprotein E, but Not Apolipoprotein B, Is Essential for Efficient Cell-to-Cell Transmission of Hepatitis C Virus. (2015)
  • Author(s): Gondar Virginia, Molina-Jimenez Francisca, Hishiki Takayuki, Garcia-Buey Luisa, Koutsoudakis George, Shimotohno Kunitada, Benedicto Ignacio, Majano Pedro.
  • Journal Title: J Virol Volume: 89(19) Issue: 19 Pages: 9962-73
  • DOI: 10.1128/jvi.00577-15
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Murine Herc6 Plays a Critical Role in Protein ISGylation In Vivo and Has an ISGylation-Independent Function in Seminal Vesicles. (2014)
  • Author(s): Arimoto KI, Hishiki T, Kiyonari H, Abe T, Cheng C, Yan M, Fan JB, Futakuchi M, Tsuda H, Murakami Y, Suzuki H, Zhang DE, Shimotohno K
  • Journal Title: J Interferon Cytokine Res. Volume: 35 Issue: 5 Pages: 351-358
  • DOI: 10.1089/jir.2014.0113
  • Peer Reviewed
• [Presentation] 不飽和脂肪酸合成酵素阻害剤によるデングウイルス複製抑制機構の解析 (2017)
  • Author(s): 日紫喜隆行、加藤文博、仁尾泰徳、宮崎恭行、田島茂、土方誠、高崎智彦
  • Organizer: 第65回 日本ウイルス学会学術集会
• [Presentation] デングウイルス感染症 (2017)
  • Author(s): 日紫喜隆行
  • Organizer: 第160回 日本獣医学会学術集会
  • Invited
• [Presentation] Development of dengue virus replicon cells expressing secretory luciferase (2017)
  • Author(s): Takayuki Hishiki, Fumihiro Kato, Shigeru Tajima, Tomohiko Takasaki
  • Organizer: 2nd International Meeting on Arboviruses and their Vectors
  • Int'l Joint Research
• [Presentation] 分泌型ルシフェラーゼ発現レプリコン細胞を用いた抗デングウイルス化合物の探索 (2017)
  • Author(s): 日紫喜隆行、加藤文博、田島茂、高崎智彦
  • Organizer: 第52回 日本脳炎ウイルス生態学研究会
• [Presentation] Development of dengue virus replicon cells expressing secretory luciferase (2016)
  • Author(s): Takayuki Hishiki, Fumihiro Kato, Shigeru Tajima, Tomoyuki Miura, Tatsuhiko Igarashi, Tomohiko Takasaki, Michinori Kohara
  • 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
• [Presentation] Bromocriptine is a novel inhibitor of dengue virus replication (2016)
  • Author(s): Takayuki Hishiki, Fumihiro Kato, Yuki Ishida, Shinya Oishi, Satoru Watanabe, Subhash G. Vasudevan, Shigeru Tajima, Tomohiko Takasaki, Tomoyuki Miura, and Tatsuhiko Igarashi
  • Organizer: 4th Antivirals Congress
  • Place of Presentation: Barcelona (Spain)
  • Year and Date: 2016-09-18
  • Int'l Joint Research
• [Presentation] デングウイルス1型レポーターレプリコン細胞の樹立 (2016)
  • Author(s): 日紫喜隆行、加藤文博、田島茂、三浦智行、五十嵐樹彦、高崎智彦
  • Organizer: 第51回日本脳炎ウイルス生態学研究会
  • Place of Presentation: ホテルリステル猪苗代（福島県耶麻郡）
  • Year and Date: 2016-05-13
• [Presentation] 分泌型ルシフェラーゼを発現するデングウイルスレプリコン細胞の樹立 (2016)
  • Author(s): 日紫喜隆行、加藤文博、田島茂、三浦智行、五十嵐樹彦、高崎智彦、小原道法
  • Organizer: 第23回トガフラビペスチウイルス研究会
  • Place of Presentation: 北海道大学（北海道札幌市）
• [Presentation] Bromocriptineによる抗デングウイルス活性機構の解析 (2015)
  • Author(s): 加藤文博、石田裕樹、大石真也、藤井信孝、田島茂、高崎智彦、三浦智行、五十嵐樹彦、日紫喜隆行
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
• [Presentation] デングウイルス感染霊長類モデル構築に向けた基盤研究 (2015)
  • Author(s): 日紫喜隆行、石田裕樹、加藤文博、川上朗彦、高崎智彦、三浦智行、五十嵐樹彦
  • Organizer: 第63回日本ウイルス学会学術集会
  • Place of Presentation: 福岡国際会議場（福岡県博多市）
  • Year and Date: 2015-11-22
• [Presentation] Generation of novel recombinant DENV for development of non-human primate model (2015)
  • Author(s): 加藤文博、田島茂、高崎智彦、三浦智行、五十嵐樹彦、日紫喜隆行
  • Organizer: 第63回日本ウイルス学会学術集会
  • Place of Presentation: 福岡国際会議場（福岡県博多市）
  • Year and Date: 2015-11-22
• [Book] Hepatitis C Virus I (2016)
  • Author(s): Hishiki Takayuki, Shimizu Yuko, Ujino Saneyuki, Nishitsuji Hironori, Shimotohno Kunitada.
  • Total Pages: 18
  • Publisher: Springer book
• [Patent(Industrial Property Rights)] 抗デングウイルス剤及びそのスクリーニング方法 (2016)
  • Inventor(s): 日紫喜隆行、加藤文博、石田裕樹
  • Industrial Property Rights Holder: 日紫喜隆行、加藤文博、石田裕樹
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-140294
  • Filing Date: 2016-07-15
  • Acquisition Date: 2016-07-29