Impaired phospholipid metabolism in glaucoma

项目概要/摘要 该提案的总体目标是更好地了解小梁中受损的磷脂代谢 青光眼中的网状（TM）组织。这里的术语磷脂 (PL) 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）。更好地理解 前眼房中的 PL 代谢将有助于深入了解青光眼病理学并促进 制定新的疾病缓解干预策略。我们的长远目标是发展得更好 通过了解代谢的病理变化来干预青光眼的策略 特别是青光眼前房中的 PL 代谢。缺乏全面的知识 与健康房水 (AH) 或 TM 相比，患病者的 PL 变化代表了一个关键障碍 治疗和预防青光眼的进展。拟议的研究基于我们广泛的初步研究 过去七年进行的研究。 我们发现青光眼 TM 中 PS 显着减少，PE 脂质种类普遍增加 与对照相比。这是在对酶的水平和活性进行严格分析之后得出的。 全面、公正地展示它们在关键分支点的变化。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 截然不同的折叠变化，仅在酰基链长度（或结构特征）方面略有不同 对照和青光眼 TM 或 AH 之间的差异表明转运蛋白的功能可能是 青光眼TM异常。我们建议研究青光眼 TM 中选定的 PL 转运蛋白 组织。病理性 TM 组织始终表现出改变的生物物理特性，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，影响 小梁网的生物物理特性及其病理生理学的贡献。在目标 1 中我们将测试 青光眼中 PL 互变酶是否发生改变，在目标 2 中，我们将评估 PL 转运是否 青光眼中的蛋白质发生改变，最后在目标 3 中，我们将确定是否恢复特定的 PL 在患病状态下变得缺乏可以改善青光眼TM的生物物理特性。我们会 还评估了小鼠模型中生物物理特性与眼压降低的相关性。我们将使用常规 （蛋白质印迹、免疫吸附测定）和创新方法分析技术，例如动力学 采用成像质谱法的组织化学。该提案还提出了一个概念 创新之处在于青光眼TM中PL代谢障碍。拟议的预期成果 研究是鉴定 PL 代谢途径和 PL 中特定酶的畸变 调节 TM 生物物理特性的分子。这些新见解将对 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.