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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的老年人口中不可逆失明的最常见原因工业化国家是与年龄相关的黄斑变性（AMD）。变性视网膜色素上皮（RPE）细胞与氧化应激和炎症是AMD的关键标志。但是，详细的分子机制潜在的AMD在很大程度上仍然未知，并且没有有效的治疗或疾病的萎缩阶段。影响生理的氧化应激功能并导致RPE细胞的局灶性损失是一种重要因素导致AMD中的地理萎缩和视力丧失。因此意味着限制RPE中活性氧的形成可能有效预防或减少AMD患者观察到的RPE功能障碍。这个建议试图了解具有抗氧化功能的多功能蛋白DJ-1如何调节RPE细胞中的氧化应激反应。我们观察到增加 AMD供体RPE中的功能性非活性DJ-1的功能无活性量为以及由于氧化应激而导致的急性RPE变性的小鼠模型。这些发现为追求抗氧化剂的研究提供了坚实的临床基础 RPE细胞中DJ-1调节的机制。根据我们的初步数据 RPE变性由可能的机理途径推理总体低水平的天然DJ-1或翻译后修饰（PTM）损害了其功能。该项目中要研究的三个重叠区域是：1-测试假设氧化应激引起的RPE变性受抗氧化剂调节 DJ-1的功能； 2-检验RPE线粒体功能障碍的假设氧化应激受DJ-1调节； 3-测试DJ-1的治疗潜力。我们将评估DJ-1在两个氧化应激中保护RPE的有效性相关的临床前小鼠模型：急性RPE变性模型和慢性在外视网膜中产生类似AMD的病变的模型。此中使用的方法研究项目包括建立人和小鼠原发性RPE细胞培养物，免疫沉淀，免疫染色，活细胞成像，共聚焦显微镜，动物 RPE变性，扫描激光眼镜检查，光谱域光学的模型相干断层扫描和电图图。