RNAseq of uPA (PLAU) knockdown in human corneal fibroblasts

Scarring and fibrotic disease result from the persistence of myofibroblasts characterized by high surface expression of αv integrins and subsequent activation of TGFβ, however the mechanism controlling their surface abundance is unknown. We previously found that knockdown of uPA (urokinase plasminogen activator) in primary human corneal fibroblasts generated myofibroblasts with cell surface accumulation of integrin αvβ5 (Wang et al., 2012, PMID: 22470492). Genetic screening of these myofibroblasts was pursued to test for changes in the gene expression of degradation machinery (ligases or deubiquitinases) that could effect cell surface expression of integrins. We found that a subset of human deubiquitinases were increased in these myofibroblasts. Secondary screening revealed that the deubiquitinase USP10 regulates alpha v integrin protein expression.瘢痕形成与纤维化疾病的发生源于肌成纤维细胞的持续存在，这类肌成纤维细胞以高表面表达αv整合素（αv integrin）并后续激活转化生长因子β（TGFβ）为特征，但调控其细胞表面丰度的分子机制目前仍不明确。我们此前的研究发现，在人原代角膜成纤维细胞中敲低尿激酶型纤溶酶原激活物（urokinase plasminogen activator，uPA）可诱导肌成纤维细胞形成，并引发整合素αvβ5在细胞表面蓄积（Wang等，2012，PMID: 22470492）。本研究针对这些肌成纤维细胞开展遗传筛选，旨在检测可能影响整合素细胞表面表达的降解相关酶系统（泛素连接酶或去泛素化酶）的基因表达变化。我们观察到，此类肌成纤维细胞中有一组去泛素化酶的表达水平上调。后续次级筛选进一步揭示，泛素特异性蛋白酶10（USP10）可调控αv整合素的蛋白表达水平。