Mechanotransduction in corneal disorders

项目摘要 我们已经确定，转录辅激活因子和机械转换因子，雅普/TAZ， 上皮接触引导、肌成纤维细胞转化和TAZ（在人类中由WWTR1编码， 小鼠中的Wwtr1）对于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;角膜移植通常需要恢复视力，特别是对于常见疾病，Fuchs 角膜内皮营养不良（FECD）。FECD的特征包括过早的角膜内皮细胞（CEC） 后弹力层上细胞外基质（ECM）的变性和赘生物（称为guttae）的形成 膜（DM）。ECM固有的生物物理线索被广泛认为是普遍存在的和有效的调节剂 包括对压力的反应。尽管如此，仍然存在重大知识差距 关于DM在健康和疾病中的机械性质以及相关的机械传导， CEC事件。此外，大量证据表明氧化应激在FECD中起主要作用。 总之，我们假设TAZ通过细胞凋亡的变化在FECD的发生和发展中起关键作用。 信号传导、基质重塑和细胞应激反应。令人兴奋的初步数据文件， 敲除（Wwtr1-/-）小鼠CEC密度降低，细胞多形性增加，DM异常 与野生型（WT）同窝仔相比，此外，CEC对TAZ缺陷型小鼠的损伤导致 大泡性角膜病变和CEC再生减少，类似于在重度慢性FECD中观察到的情况。 这些数据表明，TAZ缺陷小鼠可能是一个重要的迟发性FECD模型，以确定 机械转导在其发病机制中的作用，并测试新的治疗以延迟疾病发作和/或 进展在这个建议中，我们利用这个模型来测试奈他舒地尔，一种rho激酶和 去甲肾上腺素转运抑制剂，最近在美国被批准用于青光眼。初步数据提示 奈他舒地尔增加TAZ缺陷小鼠的CEC再生。文献支持使用rho激酶 抑制剂在CEC再生中的作用，但据我们所知，奈他舒地尔从未在这方面进行过研究。中央 该提案的目标是：1）确定TAZ在CEC再生中的作用; 2）研究 奈他舒地尔用于CEC再生的小鼠模型更能预测人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.