Therapeutic Application of Genome Editing for Advanced Heart Failure
Project/Area Number |
16K09500
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Cardiovascular medicine
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Research Institution | Osaka University |
Principal Investigator |
Shuichiro Higo 大阪大学, 医学系研究科, 寄附講座准教授 (00604034)
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Co-Investigator(Kenkyū-buntansha) |
彦惣 俊吾 大阪大学, 医学系研究科, 准教授 (30423164)
高島 成二 大阪大学, 生命機能研究科, 教授 (90379272)
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Project Period (FY) |
2016-04-01 – 2019-03-31
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Project Status |
Completed (Fiscal Year 2018)
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Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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Keywords | 心筋症 / ゲノム編集 / 拡張型心筋症 / 心筋細胞 / 相同組み換え / 循環器 / 心不全 |
Outline of Final Research Achievements |
Although high-throughput sequencing can elucidate the genetic basis of hereditary cardiomyopathy, direct interventions targeting pathological mutations have not been established. Furthermore, it remains uncertain whether homology-directed repair (HDR) is effective in non-dividing cardiomyocytes. Here, we demonstrate that HDR-mediated genome editing using CRISPR/Cas9 is effective in non-dividing cardiomyocytes. Transduction of adeno-associated virus (AAV) containing sgRNA and repair template into cardiomyocytes constitutively expressing Cas9 efficiently introduced a fluorescent protein to the C-terminus of Myl2, independently of DNA synthesis. We sought to repair a pathological mutation in Tnnt2 in cardiomyocytes of cardiomyopathy model mice, and AAV-mediated HDR achieved precise genome correction at a frequency of ~12.5%. Thus, targeted genome replacement via HDR is effective in non-dividing cardiomyocytes, and represents a potential therapeutic tool for targeting cardiomyopathy.
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Academic Significance and Societal Importance of the Research Achievements |
拡張型心筋症は、心臓が拡大し、収縮する力が徐々に低下して心不全に至る、難治性の疾患である。ゲノム解析技術の進歩により、拡張型心筋症の発症に遺伝子変異が大きく関与することが明らかとなっている。革新的な技術であるCRISPR/Cas9ゲノム編集は、様々な疾患分野への医療応用が期待されている。今回我々は、マウス培養心筋細胞を用いて、心筋細胞においてゲノム編集を用いた正確な遺伝子編集が可能であることを証明した。更に、遺伝子変異をもった心筋症モデルマウスの培養心筋細胞において、約12.5%の効率で遺伝子変異を修復することに成功した。これらの知見は、難治性心筋症に対する治療開発につながることが期待される。
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Report
(4 results)
Research Products
(25 results)
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[Journal Article] Targeted Genome Replacement via Homology-directed Repair in Non-dividing Cardiomyocytes2017
Author(s)
Ishizu T, Higo S, Masumura Y, Kohama Y, Shiba M, Higo T, Shibamoto M, Nakagawa A, Morimoto S, Takashima S, Hikoso S, Sakata Y.
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Journal Title
Sci Rep.
Volume: 7
Issue: 1
Pages: 9363-9363
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Btg2 is a Negative Regulator of Cardiomyocyte Hypertrophy through a Decrease in Cytosolic RNA.2016
Author(s)
Masumura Y, Higo S, Asano Y, Kato H, Yan Y, Ishino S, Tsukamoto O, Kioka H, Hayashi T, Shintani Y, Yamazaki S, Minamino T, Kitakaze M, Komuro I, Takashima S, Sakata Y.
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Journal Title
Sci Rep.
Volume: 6
Issue: 1
Pages: 28592-28592
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Presentation] Rapid Evaluation of Candidate Causal Gene Mutation using High-content Image Analysis Combined with Targeted Gene Disruption2018
Author(s)
Yuki Masumura, Suzuka Kunimatsu, Shuichiro Higo, Yasuaki Kohama, Mikio Shiba, Takumi Kondo, Masato Shibamoto, Seiji Takashima, Shigeru Miyagawa, Shungo Hikoso, Yasushi Sakata
Organizer
AHA scientific sessions 2018
Related Report
Int'l Joint Research
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[Presentation] Multiplexed Measurement of Cell-type Specific Calcium Dynamics Using High-content Image Analysis2018
Author(s)
Yuki Masumura, Suzuka Kunimatsu, Shuichiro Higo, Takamaru Ishizu, Yasuaki Kohama, Mikio Shiba, Takumi Kondo, Tomoaki Higo, Masato Shibamoto, Seiji Takashima, Shungo Hikoso, Yasushi Sakata
Organizer
第82回日本循環器学会学術集会
Related Report
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