Cardiac electrophysiology evaluation system for improved drug assessment

• Project/Area Number: 22K12801
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: Osaka University
• Principal Investigator: 李 俊君 大阪大学, 大学院工学研究科, 特任准教授(常勤) (10723786)
• Co-Investigator(Kenkyū-buntansha): 劉 莉 大阪大学, 大学院医学系研究科, 特任准教授 (50380093)
• Project Period (FY): 2022-04-01 – 2025-03-31
• Project Status: Granted (Fiscal Year 2023)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2024: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2023: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: pluripotent stem cells / cardiomyocytes / maturation / regenerative medicine / Pluripotent stem cells / Cardiomyocytes / Maturation / Drug assessment / drug assessment

Outline of Research at the Start

ヒトiPS細胞由来の心筋細胞は再生医療への応用だけではなく、創薬や薬剤毒性評価への応用にも期待されている。しかし、既存の心筋細胞分化誘導法で得られた心筋細胞はまだ未熟であるため、創薬への応用に十分満たされていない。本研究課題において、自発循環進行波を利用して、未熟な心筋細胞の成熟度を加速させ、最終的に成人と近いレベルの心筋細胞を獲得し、心筋毒性評価や創薬への応用を実現することを目指している。さらに、ドラックスクリーニングの応用に合わせ、自発性循環進行波現象を誘導するデバイスを融合したMulti-wells多電極システムを開発し、ハイスループットスクリーニング可能なシステムの開発を目指す。

Outline of Annual Research Achievements

This year, we successfully integrated 2D traveling wave devices into 24-well plates for high-throughput assays. The traveling wave could be maintained stably for long periods. We also attempted to prepare a smaller device for 96-well plates. However, due to the shorter closed-loop circuit in the 96-well plate, we found that the traveling wave became unstable and disappeared after prolonged culture. Therefore, we utilized the 24-well plate device for subsequent electrophysiology assays on MEA system. iPSC-derived cardiomyocytes have been paced and matured. After a 14-day training period, the mature cardiac tissue was used for recording electrophysiological signals, which were clear and well-defined, indicating readiness for subsequent drug testing.

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

We have accomplished most of the objectives of the second year rather smoothly. We are now working according to the third-year plan.

Strategy for Future Research Activity

Next year, we will conduct multiple drug tests with varying risks of torsades de pointes (TdP) and compare the response of cardiac tissue at different stages of maturation. We will also validate the robustness and superiority of the traveling wave system in its application in drug assessment.

Research Products

• [Journal Article] Modeling Reduced Contractility and Stiffness Using iPSC-Derived Cardiomyocytes Generated From Female Becker Muscular Dystrophy Carrier (2023)
  • Author(s): Kameda Satoshi、Higo Shuichiro、Shiba Mikio、Kondo Takumi、Li Junjun、Liu Li、Tabata Tomoka、Inoue Hiroyuki、Okuno Shota、Ogawa Shou、Kuramoto Yuki、Yasutake Hideki、Lee Jong-Kook、Takashima Seiji、Ikeda Yoshihiko、Hikoso Shungo、Miyagawa Shigeru、Sakata Yasushi
  • Journal Title: JACC: Basic to Translational Science Volume: - Issue: 6 Pages: 599-613
  • DOI: 10.1016/j.jacbts.2022.11.007
  • Peer Reviewed / Open Access
• [Journal Article] Restoration of Cardiac Myosin Light Chain Kinase Ameliorates Systolic Dysfunction by Reducing Superrelaxed Myosin (2023)
  • Author(s): Hitsumoto T, Tsukamoto O, Matsuoka K, Li J, Liu L, Kuramoto Y, Higo S, Ogawa S, Fujino N, Yoshida S, Kioka H, Kato H, Hakui H, Saito Y, Okamoto C, Inoue H, Hyejin J, Ueda K, Segawa T, Nishimura S, Asano Y, Asanuma H, Tani A, Imamura R, Komagawa S, Kanai T, Takamura M, Sakata Y, Kitakaze M, Haruta JI, Takashima S.
  • Journal Title: Circulation Volume: - Issue: 25 Pages: 1902-1918
  • DOI: 10.1161/circulationaha.122.062885
  • Peer Reviewed / Open Access
• [Journal Article] Tissue Sheet Engineered Using Human Umbilical Cord-Derived Mesenchymal Stem Cells Improves Diabetic Wound Healing (2022)
  • Author(s): Zhang Jingbo、Qu Xiang、Li Junjun、Harada Akima、Hua Ying、Yoshida Noriko、Ishida Masako、Sawa Yoshiki、Liu Li、Miyagawa Shigeru
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 20 Pages: 12697-12697
  • DOI: 10.3390/ijms232012697
  • Peer Reviewed / Open Access
• [Journal Article] Engineered Tissue for Cardiac Regeneration: Current Status and Future Perspectives (2022)
  • Author(s): Li Junjun、Liu Li、Zhang Jingbo、Qu Xiang、Kawamura Takuji、Miyagawa Shigeru、Sawa Yoshiki
  • Journal Title: Bioengineering Volume: 9 Issue: 11 Pages: 605-605
  • DOI: 10.3390/bioengineering9110605
• [Presentation] Human iPSC-derived closed-loop cardiac tissue for improved drug assessment (2023)
  • Author(s): Ying Hua, Junjun Li, Xiang Qu, Jingbo Zhang, Masako Ishida, Noriko Yoshida, Hayato Miyoshi, Mikio Shiba3, Shuichiro Higo4,5, Nagako Sougawa1, Maki Takeda1, Takuji Kawamura1, Ryohei Matsuura, Toshihiko Toyofuku, Yoshiki Sawa, Li Liu and Shigeru Miyagawa
  • Organizer: The 87th Annual Scientific Meeting of the Japanese Circulation Society (JCS2023)
  • Invited