A study to elucidate influence of triplet repeat expansion on pathology in Fuchs endothelial corneal dystrophy
| Project/Area Number |
18K16947
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 56060:Ophthalmology-related
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MIYAI TAKASHI 東京大学, 医学部附属病院, 講師 (40599007)
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| Project Period (FY) |
2018-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
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| Keywords | 角膜ジストロフィ / ゲノム編集 / 角膜内皮 / Fuchs角膜内皮ジストロフィ / TCF4 / Fuchs角膜変性症 / 角膜内皮細胞 / トリプレットリピート / RNA seq / 全ゲノムシーケンス / Fuchs角膜内皮変性症 / CRISPR/Cas9 / RNAseq / フックス角膜内皮変性症 / RNA-seq |
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| Outline of Final Research Achievements |
In this research, STR assay and repeat primed PCR were performed on the DNA of 41 outpatients with Fuchs corneal endothelial dystrophy, and repeat elongation of TCF4 was observed in 8 patients (19.5%).
We succeeded in cleaving the repeat of the immortalized corneal endothelial cell HCEnC using CRISPR-Cas9, and succeeded in creating a knock-out cell line. We also succeeded in producing knock-in cell lines for HCEnC with 60-repeat and 80-repeat donors using the Homology direct repair method. RNA foci was observed in the 80 repeat knock-in cell line, and TCF4 gene expression by RT-PCR was increased.
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| Academic Significance and Societal Importance of the Research Achievements |
本課題における検討において、世界で角膜移植の約４０％を占めるFuchs角膜内皮変性症について本邦の症例のDNA解析を行い、TCF4遺伝子のリピート伸長型が一定の比率を占めることを確認できた。また、CRISPR-Cas9を用いて角膜内皮細胞株のリピートのノックアウトおよびノックインに成功し、リピート切断治療の可能性および、リピート伸長によるRNA凝集という細胞内の病態を反映するモデルを作成できた。今後のFuchs角膜内皮変性症の病態解明および治療法の検討にあたり、大変有用なモデル細胞で学術的、社会的意義があると考えられる。
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Report
(3 results)
Research Products
(18 results)
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[Journal Article] Ultraviolet A light induced DNA damage and estrogen-DNA adducts in Fuchs endothelial corneal dystrophy causing females to be more affected.2020
Author(s)
Liu C, Miyajima T, Melangath G, Miyai T, Vasanth S, Deshpande N, Kumar V, Ong Tone S, Gupta R, Zhu S, Vojnovic D, Chen Y, Rogan EG, Mondal B, Zahid M, Jurkunas UV.
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Journal Title
Proc Natl Acad Sci U S A.
Volume: 117
Issue: 1
Pages: 573-583
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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