| Project/Area Number |
17K10151
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Keio University |
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Principal Investigator |
KODO Kazuki 慶應義塾大学, 医学部(信濃町), 講師 (10338105)
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| Co-Investigator(Kenkyū-buntansha) |
芝田 晋介 慶應義塾大学, 医学部(信濃町), 講師 (70407089)
湯浅 慎介 慶應義塾大学, 医学部(信濃町), 講師 (90398628)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
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| Keywords | 心筋前駆細胞 / iPS細胞 / ゲノム編集 / 心臓領域 / 心臓前駆細胞 / 再生医学 / 発生・分化 |
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| Outline of Final Research Achievements |
In cardiac development, it is known that the left and right ventricular cardiomyocytes are derived from different progenitor cells. However, the protocol for inducing and isolating different types of human cardiac progenitor cells from pluripotent stem cells has not been elucidated. We aimed to develop a system that expresses a fusion protein between a fluorescent protein and two cardiac transcription factors. Each cardiac transcription factor is specific to each cardiac progenitor cell that supplies the right or left ventricular myocardium.
The target reporter gene was successfully knocked-in to three targeted gene regions, NKX2.5, ISL1, and PPP1R12C. However, the system using a binding peptide called P2A did not work, and instead of P2A peptide, it was recommended to use of the IRES2 system, which expresses two different mRNAs simultaneously using the same promoter.
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| Academic Significance and Societal Importance of the Research Achievements |
心室特異的な心筋細胞発生の分子・細胞レベルでの機序解明および心室特異的心筋細胞の分化誘導法の確立は、左心室と右心室の心筋の性質的差異を明らかにすることによる、心室特異的な心不全治療法開発への貢献、再生医療の実現に向けて強力な知見を提供できるインパクトを持つ。今回の研究機関に、ヒトiPS細胞から分化誘導した心筋前駆細胞の単離に用いる蛍光タンパクについて、従来広く用いられてきたP2Aペプチドを用いた融合タンパクの発現が適さないことが明らかとなった。今後IRES2という、2種のmRNAを同時に生成する方法を用いたシステムを用いることにより、左右心室特異的な心臓前駆細胞単離の実現が期待される。
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