| Project/Area Number |
16K11147
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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 |
Obstetrics and gynecology
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| Research Institution | Osaka City University |
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Principal Investigator |
Koyama Masayasu 大阪市立大学, 大学院医学研究科, 教授 (00183351)
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| Co-Investigator(Kenkyū-buntansha) |
三杉 卓也 大阪市立大学, 大学院医学研究科, 講師 (30761605)
橘 大介 大阪市立大学, 大学院医学研究科, 准教授 (50381992)
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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 |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥130,000 (Direct Cost: ¥100,000、Indirect Cost: ¥30,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | 骨盤臓器脱 / 細胞外マトリックス / 子宮脱 / 膀胱瘤 / fibulin-5 / ノックアウトマウス / 幹細胞移植 / fibulin 5 / 結合組織 / 直腸脱 / CRISPR-Cas9 / fibulin-5/DANCE |
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| Outline of Final Research Achievements |
Focusing on extracellular matrix proteins (ECM), we found that expression of extracellular matrix-associated protein genes (fibulin-5, ITG) decreased as pelvic organ prolapse (POP) progressed, using human specimens from POP. The molecular mechanism of POP progress was related abnormality of ECM. Based on this knowledge, knockout mice of fibulin-5 was made using CRISPR Cas9 system. Guide RNA targeting the mouse fibulin-5 gene was prepared, microinjected into mouse embryos along with Cas9 protein and implanted into the uterus of pseudo pregnant mice. DNA sequence revealed a mutation in the target gene sequence in approximately 50% of the mice. Homozygotes developed POP after delivery in mice. HE stains showed thinning of the vaginal wall and elastic fibers around the vaginal wall tended to decrease. However, because the maintenance of homozygous mice is unstable, the possibility of gene therapy is currently being examined by preparing mesenchymal stem cell stem cells.
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| Academic Significance and Societal Importance of the Research Achievements |
高齢化や妊娠・分娩は骨盤底臓器の弛緩を惹起し、骨盤臓器脱や下部尿路症状は女性の生活の質を低下させる。骨盤臓器脱は分娩や外傷などの物理的誘発因子と組織学的な骨盤底の結合組織や細胞外マトリックスの変化を伴う遺伝的な因子などの複合的な疾病であるが、前者の疫学的研究は進んでいるが、後者の因子の解明は進んでいない。骨盤底支持機構における細胞外マトリックスの蛋白レベルの研究は本分野の解明に大きく寄与すると考えられる。本研究ではノックアウトマウスを用いた骨盤臓器脱発症に関連する蛋白のリモデリングの解析を行い、骨盤臓器脱を個別化し、幹細胞移植をターゲットとした治療を開発することが可能と考えられる。
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