Proteomic Analyses of Human Trabecular Meshwork

描述(申请人提供)：青光眼是指一组分子基础知之甚少的眼部疾病。除“正常眼压性”青光眼外，这些疾病都与眼压升高有关。小梁网(TM)对房水流出的抵抗力增加在原发性开角型青光眼(POAG)的发生和发展中起着关键作用。该方案的假设是POAG相关蛋白质和蛋白质修饰有助于阻断TM中的房水流出。对活体小梁切除的TM组织和角膜移植遗留的边缘组织进行蛋白质组学分析，以确定正常和青光眼供者之间的蛋白质差异。液相色谱、串联质谱学和生物信息学方法将用于鉴定POAG相关蛋白质和蛋白质修饰。在初步研究中，4-羟基壬烯醛(HNE)和精嘧啶氧化蛋白修饰似乎在青光眼组织中更为普遍。此外，Cochlin是一种与遗传性耳聋有关的蛋白质，只在青光眼组织中发现。Western分析和免疫组织化学将被用来探索正常和青光眼TM组织中蛋白质修饰的差异，并验证青光眼TM组织中HNE修饰的普遍性。此外，还将使用Western分析和免疫组织化学方法来验证Cochlin与青光眼小梁网状斑块/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，并促进有效治疗方法的发展，以限制疾病的进展。

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.