Impaired phospholipid metabolism in glaucoma

项目总结/摘要 本提案的总体目标是更好地了解骨小梁中受损的磷脂代谢， 青光眼中的网状（TM）组织。术语磷脂（PL）在此主要指三类脂质： 磷脂酰胆碱（PC）、磷脂酰丝氨酸（PS）、磷脂酰乙醇胺（PE）。更好地了解 前房中的PL代谢将提供对青光眼病理学的了解，并促进青光眼的发生。 制定新的、改变疾病的干预战略。我们的长远目标是发展得更好 通过了解代谢的病理变化， 特别是PL代谢，在青光眼前房。缺乏全面的知识 与健康的房水（AH）或TM相比，患病患者的PL变化是一个关键的屏障， 青光眼的防治进展这项研究是基于我们广泛的初步研究。 过去七年进行的研究。 我们发现在昏迷性TM中PS显著减少，PE脂质种类普遍增加 与对照相比。这是在严格分析了一个人体内酶的水平和活性之后得出的。 全面和公正的方式，展示他们的变化在关键分支点。因此我们 建议以公正的方式全面研究所有PL互变酶。我们还发现 在酰基链长度（或结构特征）方面仅略有不同的折叠变化 对照组与脑梗塞TM或AH之间的差异表明转运蛋白的功能可能与脑梗塞的发病有关。 昏迷性TM异常。我们建议调查选定的PL转运蛋白在昏迷TM 组织.病理性TM组织一致地表现出改变的生物物理性质，例如升高的 弹性模量我们的中心假设是，青光眼中磷脂代谢受损， 生物物理特性的小梁网，并有助于其病理生理。在目标1中，我们将测试 PL相互转换酶是否在青光眼中改变，在目标2中，我们将评估PL转运是否 蛋白质在青光眼中发生改变，最后在目标3中，我们将确定是否恢复特定的PL， 在疾病状态下变得缺乏可以改善昏迷TM的生物物理性质。我们将 还评价了生物物理特性与小鼠模型中IOP降低的相关性。我们将使用常规 （蛋白质印迹、免疫吸附测定）和创新的分析方法，如动力学分析技术， 采用成像质谱法的组织化学。该提案还提出了一个概念性的 创新之处是PL代谢障碍在青光眼TM中的应用。拟议方案的预期成果 研究是鉴定PL代谢途径的特定酶和PL中的畸变。 这些分子使TM的生物物理特性失调。这些新的见解将对 开发新的青光眼疗法。

PROJECT SUMMARY / ABSTRACT The overall goal of this proposal is to better understand impaired phospholipid metabolism in the trabecular meshwork (TM) tissue in glaucoma. The term phospholipid (PL) here refers to primarily three classes of lipids: phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylethanolamine (PE). A better understanding of PL metabolism in the anterior eye chamber will provide insight into glaucoma pathology and facilitate the development of new, disease-modifying intervention strategies. Our long-term goal is to develop better intervention strategies for glaucoma by understanding the pathological changes in metabolism, and in particular PL metabolism, in the glaucomatous anterior chamber. The absence of a comprehensive knowledge of PL changes in diseased compared to healthy aqueous humor (AH) or TM represents a critical barrier to progress in treating and preventing glaucoma. The proposed research is based on our extensive preliminary studies performed over the past seven years. We discovered a substantial decrease in PS and a general increase in PE lipid species in glaucomatous TM compared to controls. This has followed rigorous analyses of levels and activity of enzymes in a comprehensive and unbiased manner, demonstrating their alteration at critical branch points. Thus, we propose to comprehensively investigate all PL interconversion enzymes in an unbiased manner. We also found vastly different fold changes differing only slightly with respect to acyl-chain length (or structural features) between control and glaucomatous TM or AH suggesting that the functions of transport proteins are potentially aberrant in glaucomatous TM. We propose to investigate selected PL transport proteins in glaucomatous TM tissue. Pathologic TM tissue consistently demonstrates altered biophysical properties, for example an elevated elastic modulus. Our central hypothesis is that phospholipid metabolism is impaired in glaucoma, affecting biophysical properties of the trabecular meshwork and contributing to its pathophysiology. In Aim 1 we will test whether PL interconversion enzymes are altered in glaucoma, In Aim 2 we will evaluate whether PL transport proteins are altered in glaucoma, and finally in Aim 3 we will determine whether restoring specific PLs that become deficient in the diseased state can improve the biophysical properties of glaucomatous TM. We will also evaluate correlation of biophysical properties with IOP lowering in murine models. We will use routine (Western blot, immunosorbent assays) and innovative methods analytical techniques such as kinetic histochemistry employing imaging mass spectrometry. The proposal also has brought forth a conceptual innovation that is PL metabolism impairment in TM in glaucoma. The expected outcome of the proposed research is the identification of aberrations in specific enzymes of PL metabolizing pathways and in PL molecules that dysregulate biophysical properties of TM. These new insights will have an important impact on developing novel glaucoma therapies.