Impaired phospholipid metabolism in glaucoma

项目摘要/摘要 这项建议的总体目标是更好地了解骨小梁磷脂代谢受损的情况 青光眼的网络组织(TM)。这里的术语磷脂(PL)主要指三类脂： 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 前房的PL代谢将有助于深入了解青光眼的病理机制，并促进 开发新的、可改变疾病的干预策略。我们的长远目标是发展得更好 通过了解青光眼代谢的病理变化和青光眼的干预策略 青光眼前房特殊的磷脂酶代谢。缺乏全面的知识 与健康房水(AH)或TM相比，疾病患者的PL变化是一个关键的障碍 青光眼的治疗和预防进展。建议的研究是基于我们广泛的初步研究 在过去的七年里进行的研究。 我们发现青光眼TM的PS显著降低，PE脂类物质普遍增加 与对照组相比。这是在对一种酶的水平和活性进行严格分析后得出的 全面和不偏不倚的态度，在关键分支点上展示它们的变化。因此，我们 建议以公正的方式全面研究所有的PL相互转化酶。我们还发现 非常不同的折叠变化，仅在酰链长度(或结构特征)方面略有不同 在对照和青光眼TM或AH之间，提示运输蛋白的功能潜在地 青光眼TM异常。我们建议研究青光眼TM中特定的PL转运蛋白 组织。病理TM组织持续显示生物物理特性改变，例如隆起 弹性系数。我们的中心假设是，青光眼患者的磷脂代谢受损，影响 小梁网的生物物理特性及其对其病理生理学的贡献。在目标1中，我们将测试 在目标2中，我们将评估在青光眼中PL转换酶是否发生改变 在青光眼中，蛋白质会发生改变，最后，在目标3中，我们将确定是否恢复特定的 在疾病状态下变虚可以改善青光眼TM的生物物理特性。我们会 还评估了生物物理特性与小鼠模型眼压降低的相关性。我们会用常规的方式 (Western印迹、免疫吸附分析)和创新方法分析技术，如动力学 采用成像质谱学的组织化学。该提案还提出了一个概念性的 创新之处在于青光眼患者TM的PL代谢受损。建议的预期结果 研究鉴定磷脂酶代谢途径和磷脂酶中的特定酶的异常 不调节TM生物物理性质的分子。这些新见解将对以下方面产生重要影响 开发新的青光眼治疗方法。

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.