Impaired phospholipid metabolism in glaucoma

青光眼中磷脂代谢受损

• 批准号: 10577808
• 负责人: Sanjoy K Bhattacharya
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10577808
• 关键词: Affect; Anterior; Aqueous Humor; Atomic Force Microscopy; Biological Assay; Cadaver; Carrier Proteins; Cell membrane; Cells; Data; Development; Diagnosis; Disease; Elasticity; Enzyme-Linked Immunosorbent Assay; Enzymes; Etiology; Evaluation; Exclusion; Filopodia; Functional disorder; Gender; Generations; Glaucoma; Goals; Histocytochemistry; Human; Immunosorbents; Impairment; In Vitro; Injury; Integral Membrane Protein; Intervention; Kinetics; Knowledge; Lateral; Lecithin; Length; Lipids; Location; Mammals; Mediating; Membrane; Membrane Lipids; Metabolic; Metabolic Pathway; Metabolism; Methods; Modulus; Mus; Outcome; Pathologic; Pathology; Pathway interactions; Patients; Phosphatidylethanolamine; Phosphatidylserines; Phospholipid Metabolism; Phospholipid Transfer Proteins; Phospholipids; Physiologic Intraocular Pressure; Prevention; Primary Open Angle Glaucoma; Proteins; Research; Resistance; Sampling; Signal Transduction; Stretching; Techniques; Testing; Therapeutic; Tissues; Trabecular meshwork structure; Western Blotting; Work; analytical method; anterior chamber; biophysical properties; cellular transduction; comparison control; enzyme activity; experience; eye chamber; improved; innovation; insight; lipid metabolism; mass spectrometric imaging; microscopic imaging; mouse model; novel; phosphoethanolamine; pressure; prevent

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.

项目摘要 /摘要 该提案的总体目标是更好地了解小梁中的磷脂代谢受损 青光眼中的网状（TM）组织。这里的磷脂一词（PL）主要是指三类脂质： 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 前眼腔中的PL代谢将提供对青光眼病理学的见解，并促进 开发新的，疾病改良的干预策略。我们的长期目标是更好地发展 通过了解新陈代谢的病理变化以及在中间的干预策略 特定的PL代谢，在青光眼前腔中。缺乏全面知识 与健康的水性幽默（AH）或TM相比，患病的PL变化代表了关键的障碍 治疗和预防青光眼方面的进展。拟议的研究基于我们广泛的初步 在过去的七年中，研究进行了。 我们发现PS大幅下降，青光眼TM中PE脂质物种的普遍增加 与对照组相比。这是对酶在A中的水平和活性进行严格的分析之后的 全面而公正的方式，证明了它们在关键分支点的改变。因此，我们 建议以公正的方式全面研究所有PL互转酶。我们还发现 相对于酰基链长度（或结构特征），倍数差异很大的变化仅略有不同 在对照和青光眼TM或AH之间，表明转运蛋白的功能潜在 青光眼TM中异常。我们建议研究选定的青光眼TM中选定的PL转运蛋白 组织。病理TM组织始终显示出改变的生物物理特性，例如 弹性模量。我们的核心假设是，青光眼中的磷脂代谢受损，影响 小梁网的生物物理特性并有助于其病理生理学。在AIM 1中，我们将测试 PL互转酶是否在青光眼中发生改变，在AIM 2中，我们将评估PL转运是否转运 蛋白质在青光眼中发生了改变，最后在目标3中，我们将确定是否还原特定的PLS 在患病状态下变得不足可以改善青光眼TM的生物物理特性。我们将 还评估了鼠模型中生物物理特性与IOP降低的相关性。我们将使用例程 （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。该提议还提出了一个概念 创新是青光眼中TM中代谢损害的创新。提议的预期结果 研究是鉴定PL代谢途径和PL中的特定酶的畸变 TM的生物物理特性失调的分子。这些新见解将对 发展新型青光眼疗法。