Mitochondrial Oxidative Stress in the Retinal Pigment Epithelium as a Model for A

视网膜色素上皮中的线粒体氧化应激作为 A 的模型

• 批准号: 8233302
• 负责人: Alfred S Lewin
• 金额: $ 54.24万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8233302
• 关键词: Adult; Affect; Age related macular degeneration; Aging; Alleles; Antioxidants; Apoptosis; Area; Atrophic; Behavioral; Blindness; Bruch' s basal membrane structure; Catalytic RNA; Cell physiology; Cells; Choroidal Neovascularization; Complement; Country; DNA; Disease; Elderly; Electrophysiology (science); Elements; Energy Metabolism; Enzyme Gene; Enzymes; Exhibits; Exudative age-related macular degeneration; Eye; Fluorescein Angiography; Functional disorder; Fundus; Gene Transfer; Genes; Genetic Predisposition to Disease; Glutathione S-Transferase; Goals; Human; Image; Inflammation Process; Inflammatory; Knock-out; Lead; Learning; Life; Lipofuscin; Localized Disease; Measures; Mitochondria; Modeling; Monitor; Mus; Mutation; NAD(P)H dehydrogenase (quinone) 1, human; Neonatal; Neural Retina; Nonexudative age-related macular degeneration; Optical Coherence Tomography; Oral; Oxidative Stress; Pathologic; Pathology; Pathway interactions; Patients; Peripheral; Pharmaceutical Preparations; Photoreceptors; Play; Process; Proteins; RNA; Reactive Oxygen Species; Resolution; Response Elements; Retina; Retinal; Retinal Cone; Retinal Degeneration; Role; SOD2 gene; Sampling; Site; Source; Staging; Structure of retinal pigment epithelium; Susceptibility Gene; System; Testing; Time; Transcription factor genes; United States; Up-Regulation; Vascular Endothelial Growth Factors; Viral; adduct; base; effective therapy; geographic atrophy; heme oxygenase-1; human SOD2 protein; improved; in vivo; inhibitor/antagonist; macula; morphometry; mouse model; novel; oxidative damage; prevent; public health relevance; recombinase; research study; response; retinal damage; stem; transcription factor

DESCRIPTION (provided by applicant): Age Related Macular Degeneration (AMD) is a major blinding disease of the elderly in this country and, while VEGF inhibitors often stem the late, exudative stages of the disease, there is no effective therapy for the earlier, atrophic form of AMD. Oxidative damage to the retinal pigment epithelium (RPE) contributes to AMD by reducing the function of these cells and by stimulating an inflammatory cascade that leads to the pathologic hallmarks of the disease, localized atrophy of the retina and choroidal neovascularization. One source of the RPE- damaging reactive oxygen species (ROS) are the mitochondria of the RPE cells themselves. To determine whether mitochondria are an important source of ROS in the RPE in vivo, the SOD2 gene, encoding the protective enzyme MnSOD, will be deleted specifically in the RPE of mice, using a mouse line containing an allele of SOD2 flanked by loxP sites and RPE-specific expression of Cre recombinase. We plan 3 sets of experiments: (1) We will monitor the time course of retinal degeneration in real time in living mice using electrophysiology (ERG), high resolution structural analysis (SD-OCT) and behavioral analysis (Optomotry). In post mortem samples, we will examine morphological changes to the RPE, Bruch's membrane and the neural retina and to measure accumulation of lipofuscin in the RPE. We will induce the deletion of SOD2 both in neonatal and in adult mice to determine which approach better models human geographic atrophy. (2) We will increase mitochondrial oxidative stress by deletion of SOD2 in mice bearing a homozygous mutation in the gene for the transcription factor Nrl. These mice have a cone-only retina which may resemble the cone-rich human macula better than the normal mouse retina. By this means we hope to learn why the central retina is more sensitive to oxidative stress than the peripheral retina. (3) We will attempt to counteract mitochondrial oxidative stress by elevating a set of antioxidant enzymes, including heme oxygenase-1, glutathione transferases, and NAD(P)H:quinone oxidoreductase 1. These genes for these enzymes contain a common sequence component denoted "ARE" for antioxidant sequence element. We will stimulate this antioxidant response in two ways-by viral delivery of a gene that stimulates the response and by oral delivery of a novel drug that activates this pathway. We hope that these approaches may lead to a therapy for the atrophic form of AMD. PUBLIC HEALTH RELEVANCE: Age related macular degeneration (AMD) is the leading cause of blindness among the elderly in the United States. It is caused by degradation of the central retina, an area called the macula. There are two forms of the disease: atrophic or "dry" AMD and exudative or "wet" AMD. This project seeks to establish a mouse model of the pathology related to AMD and to use this model to test potential therapies for the atrophic form of the disease.

描述（由申请人提供）：与年龄相关的黄斑变性（AMD）是该国老年人的主要盲目疾病，尽管VEGF抑制剂通常会阻止该疾病的晚期，渗出性的阶段，但对早期的萎缩形式AMD没有有效的治疗。视网膜色素上皮（RPE）对氧化损伤通过降低这些细胞的功能并刺激导致疾病的病理标志的炎症性级联反应来促进AMD，从而局部的视网膜萎缩和脉络膜新生血管内化。 RPE的活性氧（ROS）的一个来源是RPE细胞本身的线粒体。为了确定线粒体在体内RPE中是否是ROS的重要来源，编码保护性酶MNSOD的SOD2基因将使用包含LOXP位点侧翼的SOD2等位基因的小鼠系列的小鼠系列中专门删除在小鼠的RPE中，并以CRE的表达和CRE的表达。我们计划了3组实验：（1）我们将使用电生理学（ERG），高分辨率结构分析（SD-OCT）和行为分析（验光）（验光）来监测活小鼠的视网膜变性的时间过程。在验尸样品中，我们将检查RPE，BRUCH膜和神经视网膜的形态变化，并测量脂肪霉素在RPE中的积累。我们将在新生儿和成年小鼠中诱导SOD2的缺失，以确定哪种方法更好地模型人类的地理萎缩。 （2）我们将通过在转录因子NRL基因中带有纯合突变的小鼠中SOD2的缺失来增加线粒体氧化应激。这些小鼠的仅锥形视网膜可能比正常小鼠视网膜更好地类似于锥形的人黄斑。通过这种方式，我们希望了解为什么中央视网膜比周围视网膜更敏感。 (3) We will attempt to counteract mitochondrial oxidative stress by elevating a set of antioxidant enzymes, including heme oxygenase-1, glutathione transferases, and NAD(P)H:quinone oxidoreductase 1. These genes for these enzymes contain a common sequence component denoted "ARE" for antioxidant sequence element.我们将以两种方式刺激这种抗氧化剂反应，从而刺激一种基因，从而刺激激活该途径的新型药物的反应和口服递送。我们希望这些方法可能会导致对AMD萎缩形式的疗法。 公共卫生相关性：与年龄相关的黄斑变性（AMD）是美国老年人失明的主要原因。它是由中央视网膜降解引起的，该区域称为黄斑。该疾病有两种形式：萎缩或“干” AMD和渗出症或“湿” AMD。该项目旨在建立与AMD相关的病理学的小鼠模型，并使用该模型来测试该疾病萎缩形式的潜在疗法。