Screening of novel podocyte protective reagents using nephrin reporter mice

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K15469
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Kidney internal medicine
• Research Institution: University of Shizuoka
• Principal Investigator: Mori Kiyoshi 静岡県立大学, 薬学部, 特任教授 (60343232)
• Co-Investigator(Kenkyū-buntansha): 松阪 泰二 東海大学, 付置研究所, 助教授 (50317749)
• Co-Investigator(Renkei-kenkyūsha): OHTSUKA Masato 東海大学, 医学部, 准教授 (90372945)
• Research Collaborator: TSUCHIDA Junichi 京都大学, 医学部, 研究員 (10643570)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: 腎臓内科学 / 糸球体 / ポドサイト / 化合物ライブラリー / スクリーニング / レポーターアッセイ

Outline of Final Research Achievements

Nephrin is a critical component of glomerular filtration barrier. Downregulation of nephrin expression is commonly observed at early stage of glomerular disorders, suggesting that methods to increase nephrin expression in podocytes may have therapeutic utility. Here, we generated a knockin mouse line carrying single copy of 5.5 kb nephrin promoter controlling expression of enhanced green fluorescent protein (EGFP) at Rosa26 genomic locus (Nephrin-EGFP mouse). Next, we isolated and cultivated glomeruli from these mice, and developed a protocol to automatically quantitate EGFP expression in cultured glomeruli. EGFP signal was markedly reduced after 5 days of culture but reduction was inhibited by vitamin D treatment. Thus, we generated a mouse line converting nephrin promoter activity into fluorescence, which can be used to screen compounds having activity to enhance nephrin gene expression.

Free Research Field

腎臓内科学