Molecular Analysis of the Human Aqueous Outflow Pathway

人体房水流出途径的分子分析

• 批准号: 7539893
• 负责人: MICHAEL P. FAUTSCH
• 金额: $ 32.33万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7539893
• 关键词: Adherent Culture; Affect; Animal Model; Anterior; Appearance; Aqueous Humor; Biochemical; Cell Culture Techniques; Cells; Ciliary Body; Culture Media; Culture Techniques; Development; Diffusion; Environment; Event; Extracellular Matrix Proteins; Eye; Gene Expression; Gene Proteins; Glaucoma; Goals; Growth Factor; Homeostasis; Human; In Vitro; Iris; Modeling; Molecular; Molecular Analysis; Molecular Profiling; Nutrient; Organ; Organ Culture Techniques; Pathway interactions; Patients; Physiological; Plant Roots; Plasma Proteins; Primary Open Angle Glaucoma; Process; Proteins; RNA; Regulation; Research; Research Personnel; Resistance; Resistance development; Serum; Serum-Free Culture Media; Source; Study models; Techniques; Tissue Culture Techniques; Tissues; Trabecular meshwork structure; aqueous; cytokine; in vivo; programs; protein expression; protein profiling

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.

我们研究计划的目标是识别与水流出阻力有关的蛋白质，以及 表征青光眼中蛋白质的改变是否会影响房水流出阻力。 由房水中的蛋白质以及小梁细胞产生的蛋白质和细胞外基质组成。 由于不存在原发性开角型青光眼 (POAG) 的动物模型，因此对人类房水的研究 流出通路在过去的30年里一直使用细胞和组织培养技术来分析小梁 这些技术使用与正常网络不同的培养基补充剂（血清等）。 房水环境。我们的研究发现当前的培养条件改变了小梁网 与房水中培养的网状结构相比，蛋白质表达（在细胞和组织中）。 在房水中培养的小梁网保持更接近的蛋白质谱 我们假设房水中发现的蛋白质需要新鲜的、非培养的网络。 维持小梁细胞和网状结构接近其正常生理状态细胞和网状结构的研究。 不在房水中培养导致的结果可能与理解生理学无关 我们进一步探究了水性蛋白质的类型和比例的变化。 （生长因子、细胞因子等）可以改变小梁稳态，导致房水流出异常 当前的提案将研究我们的假设。 以下方式： ¿表征房水对人小梁分子和细胞活动的影响 网状培养模型并确定哪些水性蛋白质对小梁细胞很重要 体内平衡。 ¿创建从正常和 POAG 获得的房水中发现的蛋白质的全面概况 我们还将确定由患者引起的网络中的基因和蛋白质表达变化。 正常和 POAG 房水。 ¿开发一种可用于模拟房水进行小梁网细胞培养的培养基。