Proteomic Analyses of Human Trabecular Meshwork

描述（由申请人提供）：青光眼是一组分子基础尚不清楚的眼科疾病的统称。除了“正常压力”青光眼，这些疾病与眼内压升高有关。小梁网（TM）对房水流出的阻力增加似乎在原发性开角型青光眼（POAG）的发病和进展中起关键作用。该提案的假设是POAG相关蛋白和蛋白修饰有助于TM中的水流出的阻塞。将对来自体内小梁切除术的TM组织和来自角膜移植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-羟基壬烯醛（HNE）和精氨酸嘧啶氧化蛋白修饰似乎更普遍的肿瘤组织。此外，与遗传性耳聋相关的蛋白质cochlin仅在昏迷组织中发现。Western分析和免疫组织化学将用于探测正常和脑肿瘤TM组织之间蛋白质修饰的差异，并验证脑肿瘤TM组织中HNE修饰的普遍性。Western分析和免疫组织化学也将用于检验cochlin与青光眼小梁网斑块/粘多糖沉积相关的假设。该提案的长期目标是更好地了解青光眼发病机制，并促进开发有效的治疗方法以限制疾病进展。

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.