Regulation of the Nrf2-mediated Antioxidant Defense In Diabetic Retinopathy

糖尿病视网膜病变中 Nrf2 介导的抗氧化防御的调节

• 批准号: 9908331
• 负责人: William P Miller
• 金额: $ 3.22万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9908331
• 关键词: 5' Untranslated Regions; Ablation; Adaptor Signaling Protein; Address; Adult; Affect; Age; American; Antioxidants; Attenuated; Award; Barbering; Binding; Biological Models; Blindness; Cell Culture Techniques; Cell Death; Cells; Complications of Diabetes Mellitus; DNA Damage; Data; Defect; Development; Diabetes Mellitus; Diabetic Retinopathy; Diabetic mouse; Disease; Down-Regulation; Electroretinography; Environment; Event; Exclusion; Exhibits; Experimental Models; Fellowship; Free Radicals; Functional disorder; Gene Expression; Genes; Genetic; Genetic Translation; Goals; Human; Hyperglycemia; Impairment; Individual; Innovative Therapy; Insulin; Intervention; Knock-out; Knockout Mice; Laboratories; Lead; Link; Maintenance; Mediating; Medicine; Messenger RNA; Metabolic; Modeling; Molecular; Muller' s cell; Mus; NF-E2-related factor 2; Neuroglia; Nuclear; Optical Coherence Tomography; Outcome; Outcome Study; Oxidative Stress; Pathogenesis; Pathology; Pathway interactions; Phosphorylation; Phosphotransferases; Polyubiquitination; Preventive Intervention; Production; Protein Dephosphorylation; Protein phosphatase; Proteins; Protocols documentation; RNA-Binding Proteins; Reactive Oxygen Species; Regulation; Retina; Retinal Defect; Ribosomes; Role; Severities; Signal Transduction; Site; System; Systems Analysis; Technical Expertise; Technology; Testing; Therapeutic; Therapeutic Intervention; Tissues; Training; Translations; Universities; Vision; Visual; beta-Transducin Repeat-Containing Proteins; biological adaptation to stress; college; design; diabetic; impaired capacity; in vivo; interest; novel; nuclear factor-erythroid 2; prevent; protein expression; public health relevance; recruit; response; retinal neuron; skills; targeted treatment; theories; training opportunity; transcription factor; ubiquitin-protein ligase; virtual; visual threshold

Project Summary Diabetic retinopathy is the leading cause of blindness in working age Americans, affecting more than a third of the ~20 million individuals with diabetes. The pathogenesis of this disease is defined by a combination of hyperglycemia and a reduction in insulin mediated signaling, which impacts retinal neurons, glia, and vasculature. A unifying theory for the pathophysiology of diabetic complications suggests that the principle pathways responsible for hyperglycemia-induced tissue damage are all linked to the accumulation of reactive oxygen species (ROS). In diabetes, the retina exhibits an increase in the production of ROS and an impaired capacity to reduce free radicals. My central hypothesis is that diabetes-induced expression of the stress response protein regulated in development and DNA damage 1 (REDD1) inhibits the nuclear factor erythroid- 2-related factor 2 (Nrf2)-related antioxidant response in the retina, leading to increased oxidative stress and retinal pathology. In support of my hypothesis, I present compelling preliminary data demonstrating that diabetes-induced oxidative stress is attenuated in the retina of REDD1 knockout mice. In addition, expression of Nrf2-responsive mRNAs is increased in the retina of REDD1 knockout mice and nuclear Nrf2 protein expression and activity are enhanced in REDD1 knockout human MIO-M1 retinal cells in culture. I plan to test my central hypothesis by pursuing the following specific aims: 1. Establish the impact of REDD1 on Nrf2 synthesis in experimental models of diabetes. 2. Delineate the impact of REDD1 on Nrf2 degradation in experimental models of diabetes. To test my hypothesis, I will pursue an experimental protocol involving model systems ranging from cell culture to intact mice, as well as cutting-edge technologies for analyzing mRNA translation. This fellowship award will also provide two key training opportunities. First, I will train with Dr. Sui Wang (Stanford University) to develop skills necessary to manipulate gene expression in the retina in vivo. Subsequently, I will receive training from Dr. Alistair Barber (Penn State College of Medicine) to develop the technical expertise to assess the impact of diabetes on retinal pathophysiology using optical coherence tomography, electroretinograms, and virtual optomotry. With respect to outcomes, this project will not only expand my skills and systems of analysis, but will also identify novel mechanisms that link the molecular events caused by the diabetic metabolic environment to the development of retinal pathology. Identification of such mechanisms is significant because it will validate new targets for the development of preventive and/or therapeutic interventions aimed at addressing the molecular basis of diabetic retinopathy and promoting healthy vision.

项目概要 糖尿病视网膜病变是工作年龄美国人失明的主要原因，影响了超过三分之一的人 约 2000 万糖尿病患者。这种疾病的发病机制是由以下因素的组合定义的： 高血糖和胰岛素介导的信号传导减少，这会影响视网膜神经元、神经胶质细胞和 脉管系统。糖尿病并发症病理生理学的统一理论表明，原则 导致高血糖引起的组织损伤的途径都与反应性物质的积累有关 氧物质（ROS）。在糖尿病中，视网膜表现出 ROS 产生增加和受损 减少自由基的能力。我的中心假设是糖尿病引起的压力表达 发育和 DNA 损伤调节反应蛋白 1 (REDD1) 抑制核因子红系- 视网膜中与 2 相关因子 2 (Nrf2) 相关的抗氧化反应，导致氧化应激增加 视网膜病理学。为了支持我的假设，我提供了令人信服的初步数据，证明 糖尿病引起的氧化应激在 REDD1 敲除小鼠的视网膜中减弱。另外，表达 REDD1 敲除小鼠视网膜中的 Nrf2 响应 mRNA 和核 Nrf2 蛋白增加 在培养的 REDD1 敲除人 MIO-M1 视网膜细胞中，表达和活性得到增强。我计划 通过追求以下具体目标来检验我的中心假设： 1. 确定 REDD1 对 Nrf2 的影响 糖尿病实验模型中的合成。 2. 描述REDD1对Nrf2降解的影响 糖尿病实验模型。为了验证我的假设，我将采用一个实验方案，其中涉及 从细胞培养到完整小鼠的模型系统，以及用于分析的尖端技术 mRNA翻译。该奖学金还将提供两个重要的培训机会。首先，我将训练 Sui Wang 博士（斯坦福大学）培养操纵视网膜基因表达所需的技能 体内。随后，我将接受 Alistair Barber 博士（宾夕法尼亚州立大学医学院）的培训 发展技术专业知识，利用光学评估糖尿病对视网膜病理生理学的影响 相干断层扫描、视网膜电图和虚拟验光。从成果来看，该项目将 不仅扩展了我的技能和分析系统，而且还将确定将 糖尿病代谢环境引起的分子事件对视网膜病理学发展的影响。 确定此类机制非常重要，因为它将验证开发新目标 旨在解决糖尿病视网膜病变分子基础的预防和/或治疗干预措施 并促进健康的视力。