Molecular targets of Corneal Anti-fibrosis

描述(申请人提供)：角膜失明是世界上导致视力丧失的第二大原因。裸露的角膜首先也会引起眼部感染， 化学性或冲击性损伤，并迅速发展血管生成和纤维化，最终导致疤痕形成和角膜失明。仅在美国，每年因工受伤的病例就超过5万例，而在战场上，驻伊拉克和阿富汗的美军士兵死于眼睛和面对伤害的速度是躯干受伤的四倍。为解决普遍存在的谜团 对于血管纤维化药物，我们开发了一个高通量的药物筛选平台，其中证实了含有铁蛋白A的多能抑制物(WFA)。由化学遗传学方法提供的第二个创新发现，照亮了Vimentin，一种III型中间丝(IF)蛋白，作为WFA的结合靶标。分子模拟研究已经解开了四聚体波形蛋白中的WFA结合位点，并首次显示了一个高度保守的可药物结合位点，该位点保留在III型IF成员中。这种对IF蛋白药理学的前所未有的分子洞察力与对靶基因缺陷(Vim KO)小鼠的研究相结合，产生了波形蛋白作为纤维化开关的关键调节因子。在目前提出的这项研究中，我们将调查III型IF蛋白是角膜抗纤维化的可用药靶点的假设。我们将测试三个特定的目标：(1)激活的角膜常驻细胞通过过度表达波形蛋白与角膜纤维化开关细胞结合。然后，我们将验证通过遗传和药物下调波形蛋白而发挥的抗纤维化的分子机制。此外，我们计划通过诱导小鼠的波形蛋白缺乏，将小鼠从眼部疾病无虹膜中拯救出来。(2)探讨结蛋白在角膜组织修复中的作用，证实结蛋白与波形蛋白联合应用可加重角膜纤维化，并通过基因或药物手段协同下调二者的表达，可促进创伤后角膜的恢复。(3)开发溶解性更好的WFA类似物，用于局部眼科治疗。使用这些新的WFA类似物将增强我们对基于WFA的药物开发的治疗方式的理解，以及针对角膜纤维化的III型IFs的合理性。

DESCRIPTION (provided by applicant): Corneal blindness is the second leading cause of vision loss in the world. The exposed cornea is first to also become involved in ocular infections, chemical or blast injuries, and rapidly develops angiogenesis and fibrosis, which ultimately leads to scarring and corneal blindness. In the US alone, work-related eye injuries account for over 50,000 cases per year, while on the battlefield, US soldiers in Iraq and Afghanistan succumb to eye and face injuries at four times the rate of torso injuries. To tackle the prevailing enigma for angio-fibrotic drugs, we developed a high throughput screening platform for drug discovery, from which was borne out the pluripotent inhibitor withaferin A (WFA). A second innovative discovery, afforded by the chemical genetic approach, illuminated vimentin, a type III intermediate filament (IF) protein, as the binding target of WFA. Molecular modeling studies have unraveled the WFA-binding site in tetrameric vimentin and showed, for the first time, a highly conserved druggable site that is retained in members of type III IFs. This unprecedented molecular insight into IF-protein pharmacology combined with studies on target deficient (Vim KO) mice yielded vimentin as a critical regulator of the fibrotic switch. In this current proposed study, we will investigate the hypothesis that type III IF proteins are druggable targets for corneal anti- fibrosis. We will test three specific aims: (1) Activated corneal resident cells engage the corneal fibrotic switch cells by overexpressing vimentin. We will then validate the molecular mechanism of anti-fibrosis exerted by genetic and pharmacological downregulation of vimentin. Additionally, we plan to rescue mice from the ocular disease Aniridia, by inducing vimentin deficiency in these mice. (2) Investigate the role of desmin expression in corneal tissue repair, and demonstrate that desmin combines with vimentin to exacerbate corneal fibrosis and their coordinate downregulation by genetic or pharmacological means can improve corneal recovery after traumatic injury. (3) Develop WFA analogs with improved solubility for topical ocular treatment. Use of these novel WFA analogs will enhance our understanding of therapeutic modalities for WFA-based drug development and the rational to target type III IFs in corneal fibrosis.