RPE, aging and age-related macular degeneration: the role of oxidative stress

RPE、衰老和年龄相关性黄斑变性：氧化应激的作用

• 批准号: 10164788
• 负责人: VERA L BONILHA
• 金额: $ 39.04万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10164788
• 关键词: Acute; Affect; Age related macular degeneration; Aging; Animal Model; Antioxidants; Area; Atrophic; Autophagocytosis; Blindness; Cell Culture Techniques; Cell Respiration; Cell Survival; Chronic; Clinical; Cone; Confocal Microscopy; Cysteine; DNA; DNA Damage; Data; Developed Countries; Disease; Elderly; Electroretinography; Environmental Risk Factor; Etiology; Eye; Foundations; Free Radicals; Functional disorder; Generations; Genes; Genetic Risk; Human; Immunoprecipitation; Impairment; Inflammation; Knockout Mice; Label; Lasers; Lesion; Light; Mediating; Methods; Mitochondria; Mitochondrial DNA; Modeling; Molecular; Mus; Neurodegenerative Disorders; Neurons; Ophthalmoscopy; Opsin; Optical Coherence Tomography; Organelles; Oxidative Stress; Oxides; PARK7 gene; Parkinson Disease; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Phenotype; Physiological; Population; Post-Translational Protein Processing; Principal Investigator; Process; Production; Proteins; Publishing; Reactive Oxygen Species; Reagent; Regulation; Research Project Grants; Retina; Retinal Degeneration; Role; Scanning; Solid; Stains; Stress; Structure of retinal pigment epithelium; Sulfonic Acids; System; Testing; Therapeutic; Thinness; Tyrosine 3-Monooxygenase; adduct; age related; base; biological adaptation to stress; combat; dopaminergic neuron; effective therapy; effectiveness evaluation; ezrin; geographic atrophy; in vivo; innovation; live cell imaging; metabolic rate; mitochondrial dysfunction; mouse model; oxidation; oxidative damage; pre-clinical; prevent; programs; protein aggregation; sodium iodate; therapeutic evaluation

Abstract The most common cause of irreversible blindness in the elderly population in industrialized countries is age-related macular degeneration (AMD). Degeneration of retinal pigment epithelium (RPE) cells in association with oxidative stress and inflammation is a key hallmark of AMD. However, the detailed molecular mechanisms underlying AMD remain largely unknown and no effective treatment exists for the early or late atrophic stages of the disease. Oxidative stress affecting the physiological function and leading to focal loss of the RPE cells has been suggested to be an important factor contributing to geographic atrophy and vision loss in AMD. Thus implying that limiting the formation of reactive oxygen species within the RPE may effectively prevent or reduce RPE dysfunction observed in AMD patients. This proposal seeks to understand how DJ-1, a multifunctional protein with an antioxidant function, regulates oxidative stress responses in RPE cells. We have observed increased amounts of the functional inactive sulfonic oxidized DJ-1 in the RPE of AMD donors as well as in a mouse model of acute RPE degeneration due to oxidative stress. These findings provide a solid clinical foundation for pursuing studies on the antioxidant mechanisms regulated by DJ-1 in RPE cells. Based on our preliminary data we reasoned of a possible mechanistic pathway via which RPE degeneration results from overall low levels of native DJ-1 or posttranslational modifications (PTMs) that impair its function. The three overlapping areas to be investigated in this project are: 1- To test the hypothesis that RPE degeneration due to oxidative stress is regulated by antioxidant function of DJ-1; 2- To test the hypothesis that mitochondria dysfunction of the RPE due to oxidative stress is regulated by DJ-1; 3- To test the therapeutic potential of DJ-1. We will evaluate the effectiveness of DJ-1 to protect RPE in vivo, in two oxidative stress relevant pre-clinical mice models: an acute RPE degeneration model and a chronic model that generates AMD-like lesions in the outer retina. The methods utilized in this research project include establishment of human and mouse primary RPE cell cultures, immunoprecipitation, immunostaining, live cell imaging, confocal microscopy, animal models of RPE degeneration, scanning laser ophthalmoscopy, spectral-domain optical coherence tomography and electroretinograms.

摘要 老年人不可逆失明的最常见原因是 在工业化国家最常见的是年龄相关性黄斑变性（AMD）。变性 视网膜色素上皮（RPE）细胞与氧化应激和 炎症是AMD关键标志。然而，详细的分子机制 潜在的AMD在很大程度上仍然是未知的，对于早期的AMD没有有效的治疗方法。 或疾病的晚期萎缩阶段。氧化应激对机体生理功能的影响 已经提出，视网膜色素上皮细胞的功能受损并导致视网膜色素上皮细胞的局灶性丧失是视网膜色素上皮细胞的一个重要原因。 是导致AMD地图样萎缩和视力丧失的重要因素。因此 这意味着限制RPE内活性氧物质的形成可以 有效地预防或减少在AMD患者中观察到的RPE功能障碍。这项建议 试图了解DJ-1，一种具有抗氧化功能的多功能蛋白质， 调节RPE细胞中的氧化应激反应。我们观察到， AMD供体的RPE中的功能性非活性磺酸氧化的DJ-1的量为 以及在由于氧化应激引起的急性RPE变性的小鼠模型中。这些 这些发现为研究抗氧化剂提供了坚实的临床基础。 在RPE细胞中由DJ-1调节的机制。根据初步数据， 推理出一种可能的机制途径，通过该途径，RPE变性由以下原因引起： 总体水平较低的天然DJ-1或翻译后修饰（PTM）会损害其功能 功能本项目将调查的三个重叠领域是： 氧化应激引起RPE变性受抗氧化剂调节假说 DJ-1的功能; 2-为了检验RPE的线粒体功能障碍由于 氧化应激受DJ-1调节; 3-测试DJ-1的治疗潜力。我们 将评估DJ-1在体内保护RPE的有效性，在两种氧化应激中， 相关的临床前小鼠模型：急性RPE变性模型和慢性RPE变性模型。 在视网膜外层产生AMD样病变的模型。在此过程中使用的方法 研究项目包括建立人和小鼠原代RPE细胞培养， 免疫沉淀，免疫染色，肝细胞成像，共聚焦显微镜，动物 视网膜色素上皮变性模型，扫描激光检眼镜，光谱域光学 相干断层扫描和视网膜电图。