Proteomic Analyses of Human Trabecular Meshwork

描述（由申请人提供）：青光眼是一组眼部疾病的总称，其分子基础尚不清楚。除“常压”青光眼外，其他疾病均与眼压升高有关。通过小梁网（TM）的水流出阻力增加似乎在原发性开角型青光眼（POAG）的发生和发展中起关键作用。该提案的假设是POAG相关蛋白和蛋白修饰有助于阻断TM中的水流出。将对活体小梁切除术后的TM组织和角膜移植后的剩余边缘组织进行蛋白质组学分析，以确定正常和青光眼供体之间的蛋白质差异。液相色谱串联质谱和生物信息学方法将用于鉴定POAG相关蛋白和蛋白修饰。在初步研究中，4-羟基烯醛（HNE）和argpy嘧啶氧化蛋白修饰似乎在青光眼组织中更为普遍。此外，耳蜗蛋白，一种与遗传性耳聋相关的蛋白，只在青光眼组织中被发现。利用Western分析和免疫组化技术探讨正常和青光眼TM组织中蛋白修饰的差异，验证HNE修饰在青光眼TM组织中的普遍性。Western分析和免疫组织化学也将用于验证耳蜗蛋白与青光眼小梁网斑块/粘多糖沉积相关的假设。这项建议的长期目标是更好地了解青光眼的发病机制，促进开发有效的治疗方法来限制疾病的进展。

DESCRIPTION (provided by applicant): Glaucoma refers collectively to a group of eye diseases whose molecular basis is poorly understood. Except in "normal-pressure" glaucoma, the diseases are associated with increased intraocular pressure. Increased resistance to aqueous outflow through the trabecular meshwork (TM) appears to play a key role in the onset and progression of primary open angle glaucoma (POAG). The hypothesis of the proposal is that POAG associated proteins and protein modifications contribute to blockage of aqueous outflow in the TM. Proteomic analyses of TM tissue from in vivo trabeculectomy and left over rim-tissue from cornea transplant will be pursued to determine protein differences between normal and glaucomatous donors. Liquid chromatography tandem mass spectrometry and bioinformatic methods will be used to identify POAG associated protein and protein modifications. In preliminary studies, 4-hydorxynonenal (HNE) and Argpyrimidine oxidative protein modifications appear to be more prevalent in glaucomatous tissue. Furthermore, cochlin, a protein associated with inheritable deafness, has been found only in glaucomatous tissue. Western analysis and immunohistochemistry will be used to probe for differences in protein modifications between normal and glaucomatous TM tissue and verify the prevalence of HNE modification in glaucomatous TM tissue. Western analysis and immunohistochemistry will also be used to test the hypothesis that cochlin is associated with glaucomatous trabecular meshwork plaques/mucopolysaccharide deposits. The long-term goal of this proposal is to better understand mechanisms of glaucoma pathogenesis and facilitate the development of effective therapies for limiting disease progression.