Mechanotransduction in corneal disorders

项目总结 我们已经确定，转录共激活因子和机械转导因子YAP/TAZ影响角膜 上皮接触引导，肌成纤维细胞转化，以及TAZ(由WWTR1在人类和 小鼠的WWTR1)对健康的角膜内皮至关重要。角膜疾病是导致失明的主要原因。 世界范围内；经常需要角膜移植来恢复视力，特别是对于常见的情况 内皮细胞角膜营养不良(FECD)。FECD的特征包括早熟的角膜内皮细胞(CEC) Descemet‘s上细胞外基质(ECM)的变性和过度形成 膜(DM)。ECM固有的生物物理信号被广泛认为是无处不在的、强有力的调节器 无数的细胞行为，包括它们对压力的反应。尽管如此，仍然存在着重大的知识差距。 关于糖尿病在健康和疾病中的机械特性及其相关的机械转导 CEC中的事件。此外，大量证据表明，氧化应激在FECD中起着重要作用。 总之，我们假设TAZ通过细胞变化在FECD的发生和发展中起关键作用 信号、基质重塑和细胞应激反应。令人兴奋的初步数据文件，TAZ 基因敲除(WWTR1-/-)小鼠CEC密度降低，细胞多聚体增多，并出现异常的DM 与野生型(WT)窝产仔进行比较。此外，对TAZ缺陷小鼠的CEC损伤可导致 大泡性角膜病变和CEC再生减少，类似于严重的慢性FECD。 这些数据表明，TAZ缺陷小鼠可能是FECD定义的一个重要的迟发性模型 机械转导在其发病机制中的作用及延缓发病和/或延缓疾病的新疗法的试验 进步。在这个方案中，我们利用这个模型来测试netarsudil，一种Rho-Kinase和 去甲肾上腺素转运抑制剂，最近在美国被批准用于青光眼。初步数据显示， 奈瑟地尔促进TAZ缺陷小鼠CEC再生。文献支持Rho-Kinase的使用 在CEC再生中的抑制剂，但据我们所知，netarsudil从未在这方面进行过研究。中环 这项建议的目标是：1)确定TAZ在CEC再生中的作用；2)研究TAZ在CEC再生中的作用 Netarsudil用于CEC再生的小鼠模型更能预测人类FECD。

PROJECT SUMMARY We have determined that transcriptional co-activators and mechanotransducers, YAP/TAZ, influence corneal epithelial contact guidance, myofibroblast transformation, and that TAZ (encoded by WWTR1 in humans and Wwtr1 in mice) is crucial for a healthy corneal endothelium. Corneal disease is a leading cause of blindness worldwide; corneal transplantation is often required to restore vision particularly for the common condition, Fuchs endothelial corneal dystrophy (FECD). Hallmarks of FECD include premature corneal endothelial cell (CEC) degeneration and the formation of excrescences of extracellular matrix (ECM), termed guttae, on Descemet’s membrane (DM). Biophysical cues intrinsic to ECMs are widely recognized as ubiquitous and potent modulators of myriad cell behaviors, including their response to stress. Despite this, there remains a major knowledge gap in regard to the mechanical properties of DM in health and disease and the associated mechanotransduction events in CECs. Furthermore, a large body of evidence points to oxidative stress playing a major role in FECD. Together, we hypothesize that TAZ plays a critical role in the onset and progression of FECD via changes in cell signaling, matrix remodeling, and cellular stress responses. Exciting preliminary data document that TAZ knockout (Wwtr1-/-) mice have reduced CEC density, increased cellular polymegathism, and an abnormal DM with guttae in comparison to wildtype (WT) littermates. Furthermore, CEC injury to TAZ deficient mice results in bullous keratopathy and diminished CEC regeneration similar to what is observed in severe, chronic FECD. These data suggest that TAZ deficient mice may represent an important late-onset model for FECD to define the role of mechanotransduction in its etiopathogenesis and to test new therapies to delay disease onset and/or progression. In this proposal, we utilize this model to test the efficacy of netarsudil, a rho-kinase and norepinephrine transport inhibitor, recently approved for glaucoma in the US. Preliminary data suggests that netarsudil increases CEC regeneration in TAZ deficient mice. The literature supports the use of rho-kinase inhibitors in CEC regeneration but netarsudil has never been studied in this context to our knowledge. The central goals of this proposal are to 1) determine the role of TAZ in CEC regeneration and 2) investigate the efficacy of netarsudil for CEC regeneration using murine models more predictive of human FECD.