Elucidation of molecular mechanism of corneal epithelial stem cells using functional miRNA analysis

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K11487
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Ophthalmology
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: NAKAMURA TAKAHIRO 京都府立医科大学, 医学(系)研究科(研究院), 客員講師 (30411078)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 角膜上皮 / 幹細胞 / miRNA / 再生医療

Outline of Final Research Achievements

We created a comprehensive functional miRNA profile at the single-cell level and confirmed that reproducibility of miRNA 205 was significantly upregulated in human corneal epithelial stem cells in multiple human corneal epithelial cell groups. Next, a genetically modified mouse of miRNA-205 was prepared. The miRNA-205 knockout mouse was fatal during the embryonic period due to skin hypoplasia. Morphological and cytobiological analyzes of corneal epithelium during the embryonic period and up to 5 days after birth were performed. Compared with WT, slight changes such as the degree of stratification were observed, but there were no obvious morphological or cell biological differences. These results suggest that miRNA205 has different functions in embryonic and adult corneal epithelial stem cells.

Free Research Field

眼科学

Academic Significance and Societal Importance of the Research Achievements

近年の精力的な基礎研究から、タンパク質をコードしていないnon-coding RNAの一種であるmiRNAは生体内の様々な部位に存在し、発生・分化・癌化などの生命現象や幹細胞分化制御に深く関与していることが報告されている。生体内で角膜は視覚を司る重要な組織であり、その角膜上皮幹細胞の単一細胞レベルでの網羅的な機能性miRNAプロファイルの作成の試みは、世界的に見ても初めての試みであり、極めて独創的・先駆的である。また、miRNAレベルでの遺伝子改変マウスの作成の試みは、いまだ報告が少なく、遺伝改変技術を用いた生体内での恒常性維持機構の解明、分子レベルでの機能解析に大きく貢献したと考える。