Development of a novel drug to treat corneal dystrophy by drug repositioning

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K09978
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56060:Ophthalmology-related
• Research Institution: Keio University
• Principal Investigator: SHIMMURA Shigeto 慶應義塾大学, 医学部(信濃町), 准教授 (00235780)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 疾患iPS細胞 / ドラッグ・リポジショニング

Outline of Final Research Achievements

Fuchs corneal dystrophy (FCD) is a progressive disease of the corneal endothelium of unknown etiology. This investigation sought to derive induced pluripotent stem cells (iPS) from patients with Fuchs dystrophy, and to use these cells to investigate pathophysiology of the disease, as well as to develop new drugs for therapy. Following approval by the institutional review board, iPS cells were derived from the peripheral blood of 4 FCD patients (3 Japanese, 1 Dutch), and were differentiated into corneal endothelial cells. qRT-PCR was performed for markers of differentiation, ER stress, aging and cell metabolism. Immunocytochemistry was performed for cleaved caspase 3, gamma H2AX as markers of DNA damage, and data was imaged by in cell analyzer. Flux analyzer was used to measure mitochondrial stress. We found that FCD-derived endothelial cells were prone to oxidative stress, ER stress as well as DNA damage.

Free Research Field

眼科学

Academic Significance and Societal Importance of the Research Achievements

得られたFCD患者由来角膜内皮細胞に対しqRT-PCRによる分化・小胞体ストレス・老化・代謝関連分子の解析を行った。また細胞死の指標となるCleaved caspase3 ,DNA損傷の指標となるγH2AXのfociについても免疫染色しin cell analyzerを用いて解析を行った。さらにFlux Analyzerを用いてミトコンドリアストレステストを行った。結果、FCD疾患由来角膜内皮細胞では酸化ストレス・小胞体ストレス・DNA damage response が障害されていることが明らかになった。またこれらをrescueする薬剤として新規候補薬を発見した。