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: 6692140
• 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/6692140
• 关键词: REMOVAL; LIPOFUSCIN; COMPONENT; A2E; RPE

DESCRIPTION (provided by applicant): 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 fluorescence 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 Iysosomes 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 OSphospholipids, 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.

描述（由申请人提供）：该项目将开发减少视网膜上皮（RPE）中N- retinyledene-N-返回乙基乙醇胺（A2E）量的方法。该脂肪霉素成分的过度积累有助于与年龄相关的黄斑变性（ARMD）发作。因此，任何可以从RPE中删除A2E的方法都将有助于开发治疗该疾病的潜在疗法。 为了测试从培养的RPE细胞中去除A2E的各种方法的效率，我们将使用荧光板读取器和荧光显微镜测量细胞内浓度和A2E的亚细胞位置。使用体外系统的初始结果已经确定了几种能够从磷脂膜中去除A2E的药物。随后的工作将集中于将这些药物输送到A2E亚细胞积累部位的RPE的iysosomess。 最近的证据表明，A2E会损害RPE降解源自吞噬细胞外部段的磷脂的能力。每天，RPE都会带有棚外部段颗粒，并且必须在第二天之前完成这些颗粒的消化。 I磷脂消化的任何延迟，包括总OS质量的一半以上，都将导致这些磷脂的逐渐积累。一旦实现了可以去除A2E的方法的开发，将检查这些系统在恢复A2E处理的RPE降解OS OS磷脂的能力方面的有效性。该实验室已经开发了基于粉料的脂质降解测定法。