Human RPE metabolism and metabolite transport

人类RPE代谢和代谢物转运

• 批准号: 9003668
• 负责人: Jennifer Rayming Chao
• 金额: $ 39.15万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9003668
• 关键词: Age related macular degeneration; Cell Culture System; Cells; Cellular Metabolic Process; Choroid; Clinical; Data Set; Disease; Energy Metabolism; Extracellular Structure; Failure; Goals; Homeostasis; Human; Image; Life; Light; Metabolic; Metabolism; Methodology; Mitochondria; Neural Retina; Neurons; Outcome; Oxidative Stress; Pathogenesis; Patients; Phagocytosis; Photoreceptors; Process; Recycling; Regulation; Research; Resources; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Retinoids; Role; Side; Sorsby' s fundus dystrophy; Structure of retinal pigment epithelium; Testing; Time; Tissues; Tracer; absorption; carboxylation; cytokine; induced pluripotent stem cell; novel strategies; public health relevance; visual cycle

 DESCRIPTION (provided by applicant): Energy metabolism and metabolite transport are essential for the viability and function of the retinal pigment epithelium (RPE). Understanding these processes and their perturbations in disease states is of fundamental importance. Our preliminary results show reductive carboxylation is much more prominent in RPE than in retina or other neuronal cells or tissues. We found that reductive carboxylation is deficient in RPE cells derived from induced pluripotent stem cell (iPSC) cells made from a Sorsby's Fundus Dystrophy (SFD) patient and under conditions of oxidative stress, a critical component of early pathogenesis of AMD. Because of the potential disease relevance of reductive carboxylation in RPE cells, we will examine the role of reductive carboxylation in RPE cells using advanced tracer methodology, real time mitochondrial function analysis and live imaging. We observed in a well-controlled cell culture system that human RPE preferentially exports metabolites to the retinal side. We also found that metabolite transport is impaired in SFD iPSC-derived RPE cells. We plan to systematically investigate the regulation of metabolite transport by extracellular structure and intracelluar state, and identify the mechanisms of the defective transport in SFD RPE. This proposed research will generate a reference data set of metabolites that are preferentially exported and consumed by the RPE, determine the role of reductive carboxylation in RPE cell metabolism, and reveal normal and disease-relevant changes in metabolite transport in RPE. This new information can be used to develop treatment of retinal degenerative diseases.

 描述（由申请人提供）：能量代谢和代谢物运输对于视网膜色素上皮（RPE）的活力和功能至关重要。了解这些过程及其在疾病状态下的扰动至关重要。我们的初步结果表明，还原性羧化在 RPE 中比在视网膜或其他神经元细胞或组织中更为突出。我们发现 RPE 细胞缺乏还原羧化作用 源自索斯比眼底营养不良 (SFD) 患者的诱导多能干细胞 (iPSC)，并且处于氧化应激条件下，氧化应激是 AMD 早期发病机制的关键组成部分。由于 RPE 细胞中还原羧化的潜在疾病相关性，我们将使用先进的示踪剂方法、实时线粒体功能分析和实时成像来检查 RPE 细胞中还原羧化的作用。我们在控制良好的细胞培养系统中观察到，人类 RPE 优先将代谢物输出到视网膜侧。我们还发现 SFD iPSC 衍生的 RPE 细胞中代谢物转运受损。我们计划系统地研究细胞外结构和细胞内状态对代谢物运输的调节，并确定SFD RPE中运输缺陷的机制。这项拟议的研究将生成 RPE 优先输出和消耗的代谢物的参考数据集，确定还原羧化在 RPE 细胞代谢中的作用，并揭示 RPE 中代谢物运输的正常和疾病相关变化。这一新信息可用于开发视网膜退行性疾病的治疗方法。