Molecular Analysis of the Human Aqueous Outflow Pathway

我们的研究计划的目标是确定参与水外流阻力的蛋白质， 表征蛋白质是否在青光眼眼内改变。水流出阻力可能会受到影响 通过房水中的蛋白质和小梁细胞产生的蛋白质和细胞外基质。 由于没有原发性开角型青光眼（POAG）的动物模型，因此对人房水的研究 在过去的30年中，流出途径已经使用细胞和组织培养技术来分析小梁 网状结构这些技术使用的培养基补充剂（血清等）不同于正常的网状组织。 我们的研究发现目前的培养条件改变了小梁网 蛋白质表达（在细胞和组织中），与在房水中培养的网状物相比。 在房水中培养的小梁网保持一种蛋白质谱， 新鲜的、非培养的网状物。我们假设在房水中发现的蛋白质是 维持小梁细胞和网状结构接近其正常生理状态。细胞和网络的研究 没有在房水中培养的结果可能与理解 体内情况。我们进一步假设，水溶性蛋白质的类型和比例的变化 （生长因子、细胞因子等）可改变小梁稳态，导致异常房水流出 抵抗和青光眼的发展。目前的建议将调查我们的假设， 以下方式： 表征房水对人小梁细胞中分子和细胞活性的影响 网状培养模型，并确定哪些水蛋白质对小梁细胞重要 体内平衡 创建一个全面的蛋白质分布图，发现在从正常和POAG获得的房水中 患者我们还将确定基因和蛋白质表达的变化，在网络诱导 正常和POAG房水。 开发可用于模拟小梁网细胞培养的房水的培养基。

The goal of our research program is to identify the proteins involved in aqueous outflow resistance, and characterize whether proteins are altered in the glaucomatouseye. Aqueous outflow resistance can be affected by the proteins in aqueous humor and the proteins and extracellular matrix produced by trabecular cells. Because no animal model of primary open angle glaucoma (POAG) exists, studies of the human aqueous outflow pathway over the past 30 years have used cell and tissue culture techniques to analyze the trabecular meshwork. These techniques use media supplements (serum, etc) that differ from the meshwork's normal aqueous humor environment.Our studies find that current culture conditions alter trabecular meshwork protein expression (in both cell and tissue) when compared with meshworks cultured in aqueous humor. Trabecular meshworks cultured in aqueous humor maintain a protein profile that more closely resembles fresh, non-cultured meshworks. Wehypothesize that proteinsfound in aqueous humor are required to maintain trabecular cells and meshworks near their normal physiologic state. Studies of cells and meshworks not cultured in aqueous humor result in findings that may not be physiologicallyrelevant to understanding the in vivo situation. We further hypothesize that changes in the types and proportions of aqueous proteins (growth factors, cytokines, etc.) can alter trabecular homeostasis resulting in abnormal aqueous outflow resistance and development of glaucoma. The current proposal will investigate our hypothesis in the following manner: ¿ Characterize the effect of aqueous humor on molecular and cellular activities in human trabecular meshwork culture models and determine which aqueous proteins are important for trabecular cell homeostasis. ¿ Create a comprehensive profile of proteins found in aqueous humor obtained from normal and POAG patients. We will also determine gene and protein expression changes in the meshwork induced by normal and POAG aqueous humor. ¿ Develop a medium that can be used to mimic aqueous humor for trabecular meshwork cell culture.