REMOVAL OF THE LIPOFUSCIN COMPONENT A2E FROM RPE CELLS

从 RPE 细胞中去除脂褐素成分 A2E

• 批准号: 6840785
• 负责人: Enrique J Rodriguez-Boulan
• 金额: $ 16.95万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6840785
• 关键词: REMOVAL; LIPOFUSCIN; COMPONENT; A2E; RPE

EXCEED THE SPACE PROVIDED, This project will develop methods to reduce the amounts of N-retinyledene-N-retinylethanolamine (A2E) in the retinal pigment epithelia (RPE). The excessive accumulation of this lipofuscin component contributes to the onset of age-related macular degeneration (ARMD). Thus, any methods that can remove A2E from the RPE will be useful in developing potential therapies to treat this disease. To test the efficiency of various methods to remove A2E from cultured RPE cells, we will use a fluorescence plate reader and fluorscence microscopy to measure both the intracellular concentration and the subcellular location of A2E. Initial results using an in vitro system have already identified several drugs which are capable of removing A2E from phospholipid membranes. Subsequent work will focus on delivering these drugs to the lysosomes of RPE, the site of A2E subcellular accumulation. Recent evidence suggests that A2E impairs the ability of RPE to degrade phospholipids derived from phagocytosed outer segment. Each day, the RPE is presented with shed outer segment particles, and must complete the digestion of these particles before the next day. Any delay in the digestion of the OS phospholipids, which comprises more than half of the total OS mass, will result in the gradual accumulation of these phospholipids. Once the development of methods that can remove A2E is achieved, the effectiveness of these systems in restoring the ability of A2E-treated RPE to degrade OS phospholipids will be examined. A flourescence-based lipid degradation assay has already been developed in this lab. PERFORMANCE SITE ========================================Section End===========================================

本项目将开发减少视网膜色素上皮细胞（RPE）中N-视黄基-N-视黄基乙醇胺（A2 E）含量的方法。这种脂褐素成分的过度积累有助于年龄相关性黄斑变性（ARMD）的发作。因此，任何可以从RPE中去除A2 E的方法都将有助于开发治疗这种疾病的潜在疗法。 为了测试从培养的RPE细胞中去除A2 E的各种方法的效率，我们将使用荧光读板仪和荧光显微镜来测量A2 E的细胞内浓度和亚细胞位置。使用体外系统的初步结果已经确定了几种能够从磷脂膜中去除A2 E的药物。随后的工作将集中在将这些药物递送到RPE的溶酶体，A2 E亚细胞积累的位点。 最近的证据表明，A2 E损害RPE的能力，以降解来自吞噬的外节的磷脂。每一天，RPE都呈现出脱落的外节颗粒，并且必须在第二天之前完成这些颗粒的消化。OS磷脂（占总OS质量的一半以上）消化的任何延迟将导致这些磷脂的逐渐积累。一旦开发出可以去除A2 E的方法，将检查这些系统在恢复A2 E处理的RPE降解OS磷脂的能力方面的有效性。本实验室已经开发出一种基于荧光的脂质降解测定法。性能现场=