Using zebrafish for screening of Diamond-Blackfan anemia drugs

• Project/Area Number: 15K15394
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Pediatrics
• Research Institution: University of Miyazaki
• Principal Investigator: Yoshihama Maki 宮崎大学, フロンティア科学実験総合センター, 研究員 (00381103)
• Co-Investigator(Kenkyū-buntansha): 上地 珠代 宮崎大学, フロンティア科学実験総合センター, 研究員 (10381104)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 創薬 / ゼブラフィッシュ / バイオラベリング / 造血効果 / 毒性試験 / 薬物代謝 / 化合物スクリーニング / リボソーム / 疾患モデル / ゼブラフィッシュ創薬 / 貧血

Outline of Final Research Achievements

In this study, we developed an in vivo drug evaluation system that uses zebrafish as a tool for drug discovery for Diamond-Blackfan anemia (DBA). (1) We performed a compound screening as a pilot study to identify candidate drugs for DBA treatment. (2) A biolabeling method was developed to detect drug metabolism in vivo. (3) We used CRISPR-Cas9 system to generate DBA models that carry rps19 gene mutation. The rps19 homozygous mutants showed severe anemia at 48 hours post fertilization. (4) The toxicity test revealed an increased lethality depending on the time period of compound treatments.

Research Products

• [Int'l Joint Research] University of Toledo(米国)
• [Journal Article] Identification and characterization of 5alpha-cyprinol-sulfating cytosolic sulfotransferases (Sults) in the Zebrafish (Danio rerio). (2017)
  • Author(s): Kurogi, K., Yoshihama, M., Horton, A., Schiefer, I.T., Krasowski, M.D., Hagey, L.R., Williams, F.E., Sakakibara, Y., Kenmochi, N., Suiko, M., Liu, M.-C.
  • Journal Title: The Journal of Steroid Biochemistry and Molecular Biology Volume: 174 Pages: 120-127
  • DOI: 10.1016/j.jsbmb.2017.08.005
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Analysis of the translational efficiencies of mRNAs in a zebrafish model of Diamond-Blackfan anemia (2018)
  • Author(s): Tamayo Uechi, Yukari Nakajima, Maki Yoshihama, Etsuro Ito, Yutaka Suzuki, Naoya Kenmochi
  • Organizer: 15th Diamond Blackfan Anemia International Consensus Conference
  • Int'l Joint Research
• [Presentation] Decreased translational efficiency of mRNAs required for hematopoiesis in a zebrafish model of Diamond-Blackfan anemia (2017)
  • Author(s): Tamayo Uechi, Maki Yoshihama, Yukari Nakajima, Yutaka Suzuki, Sumio Sugano, Naoya Kenmochi
  • Organizer: "Translation Machinery in Health & Disease" Gordon Research Conference
  • Place of Presentation: Galveston, USA
  • Year and Date: 2017-03-19
  • Int'l Joint Research
• [Presentation] ゼブラフィッシュを用いたダイアモンド・ブラックファン貧血発症機構の解析 (2017)
  • Author(s): 上地珠代, 吉浜麻生, 中島由香里, 鈴木穣, 菅野純夫, 伊藤悦朗, 剣持直哉
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] ゼブラフィッシュを用いたリボソーム病発症機構の解析および創薬スクリーニング (2017)
  • Author(s): 上地珠代、吉浜麻生、中島由香里、鈴木穣、伊藤悦朗、剣持直哉
  • Organizer: 第３回ゼブラフィッシュ創薬研究会
• [Presentation] in vivo analysis for drug screening using DBA zebrafish model and bio labeling to visualize drug metabolism (2017)
  • Author(s): Maki Yoshihama, Tamayo Uechi, Katstuhisa Kurogi, Yukari Nakajima, Ming-Cheh Liu, Naoya Kenmochi
  • Organizer: 第23回 小型魚類研究会
• [Presentation] リボソーム病モデルにおけるmRNAの飜訳制御と疾患発症の分子機構 (2017)
  • Author(s): 上地珠代, 吉浜麻生, 中島由香里, 鈴木穣, 伊藤悦朗, 剣持直哉
  • Organizer: 第19回日本RNA学会年会
• [Presentation] Ribosomal dysfunction and defective erythropoiesis in a zebrafish model of Diamond-Blackfan anemia (2017)
  • Author(s): Tamayo Uechi, Maki Yoshihama, Yukari Nakajima, Yutaka Suzuki, Sumio Sugano, Etsuro Ito, Naoya Kenmochi
  • Organizer: The 22nd Annual Meeting of the RNA Society
  • Int'l Joint Research
• [Presentation] ゼブラフィッシュ貧血モデルを用いた創薬スクリーニングおよび薬物代謝の in vivo 解析 (2017)
  • Author(s): 吉浜 麻生, 上地 珠代, 黒木 勝久, Ming-Cheh Liu, 剣持 直哉
  • Organizer: 平成29年度日本生化学会九州支部例会
• [Presentation] ゼブラフィッシュDBAモデルを用いた創薬のための in vivo スクリーニング (2016)
  • Author(s): 吉浜 麻生, 上地 珠代, 中島 由香里, 引間 園美, 剣持 直哉
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] ゼブラフィッシュを用いたダイアモンド・ブラックファン貧血の発症機構の解析 (2016)
  • Author(s): 上地 珠代, 中島 由香里, 吉浜 麻生, 鈴木 穣, 菅野 純夫, 剣持 直哉
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台
  • Year and Date: 2016-09-25
• [Presentation] Selected mRNA translation and ribosomopathies: analyzing the zebrafish model of congenital pure red-cell (2016)
  • Author(s): Tamayo Uechi, Yukari Nakajima, Maki Yoshihama, Yutaka Suzuki, Sumio Sugano, Naoya Kenmochi
  • Organizer: RNA2016
  • Place of Presentation: 京都
  • Year and Date: 2016-06-28
  • Int'l Joint Research
• [Presentation] Investigation of the cardiovascular toxicity of oxidized cholesterols using the zebrafish as an animal model (2016)
  • Author(s): Katsuhisa Kurogi, Maki Yoshihama, Naoya Kenmochi, Yoichi Sakakibara, Masahito Suiko, Frederick Williams, Ming-Cheh Liu
  • Organizer: American Society for Biochemistry and Molecular Biology 2016 Annual Meeting
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-04-02
  • Int'l Joint Research
• [Presentation] Aberrant translation and ribosomopathies: studying the molecular pathogenesis of human diseases using zebrafish as a model animal (2016)
  • Author(s): Tamayo Uechi, Yukari Nakajima, Maki Yoshihama, Shiori Ishida, Tsutomu Toki, Yutaka Suzuki, Sumio Sugano, Etsuro Ito, Naoya Kenmochi
  • Organizer: The International Meeting on Aquatic Model Organism for Human Disease and Toxicology Research
  • Place of Presentation: 岡崎
  • Year and Date: 2016-03-18
  • Int'l Joint Research
• [Presentation] Decreased translation efficiency of mRNAs required for hematopoiesis in a zebrafish model of Diamond-Blackfan anemia (2016)
  • Author(s): Tamayo Uechi, Yukari Nakajima, Maki Yoshihama, Tsutomu Toki, Yutaka Suzuki, Sumio Sugano, Etsuro Ito, Naoya Kenmochi
  • Organizer: The 14th Diamond Blackfan Anemia International Consensus Conference
  • Place of Presentation: Atlanta, USA
  • Year and Date: 2016-03-05
  • Int'l Joint Research
• [Presentation] ノックダウン胚を用いた化合物スクリーニングおよび薬物代謝の in vivo 解析 (2016)
  • Author(s): 吉浜 麻生, 上地 珠代, 黒木 勝久, Ming-Cheh Liu, 剣持 直哉
  • Organizer: 第2回ゼブラフィッシュ創薬研究会
  • Place of Presentation: 岐阜
• [Presentation] ゼブラフィッシュを用いたリボソーム病発症メカニズムの解明 (2015)
  • Author(s): 上地珠代, 中島由香里, 吉浜麻生, 鈴木穣, 菅野純夫, 剣持直哉
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会 合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] リボソーム病モデルを用いた発症機序の解明と化合物スクリーニング (2015)
  • Author(s): 上地珠代, 吉浜麻生, 中島由香里, 上米良美香, 剣持直哉
  • Organizer: 第1回ゼブラフィッシュ創薬研究会
  • Place of Presentation: 津
  • Year and Date: 2015-11-06
• [Presentation] Selected mRNA translation and ribosomopathies: studying the molecular pathogenesis of congenital anemia using zebrafish as a model system (2015)
  • Author(s): Tamayo Uechi, Yukari Nakajima, Maki Yoshihama, Yutaka Suzuki, Sumio Sugano, Naoya Kenmochi
  • Organizer: International Conference on Ribosome Synthesis
  • Place of Presentation: Brussels, Belgium
  • Year and Date: 2015-08-19
  • Int'l Joint Research
• [Remarks] 研究室ホームページ
  • URL: http://ribosome-labo.med.miyazaki-u.ac.jp
• [Remarks] リボソームタンパク質遺伝子データベース
  • URL: http://ribosome.med.miyazaki-u.ac.jp