Regulation of the Nrf2-mediated Antioxidant Defense In Diabetic Retinopathy

项目摘要 糖尿病视网膜病变是美国工作年龄段失明的主要原因，影响超过三分之一 约2000万糖尿病患者中。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，这影响视网膜神经元，神经胶质细胞， 脉管系统糖尿病并发症的病理生理学的统一理论表明， 负责高血糖诱导的组织损伤的途径都与反应性胰岛素的积累有关。 氧物种（ROS）。在糖尿病中，视网膜表现出ROS产生的增加和受损的视网膜细胞。 减少自由基的能力。我的主要假设是糖尿病引起的应激反应 在发育和DNA损伤中调节的反应蛋白1（REDD 1）抑制核因子红细胞- 2-相关因子2（Nrf 2）相关的视网膜抗氧化反应，导致氧化应激增加， 视网膜病理学为了支持我的假设，我提出了令人信服的初步数据表明， 糖尿病诱导的氧化应激在REDD 1敲除小鼠的视网膜中减弱。此外，表达 在REDD 1敲除小鼠的视网膜中，Nrf 2反应性mRNA的表达增加， 在培养的REDD 1敲除人MIO-M1视网膜细胞中表达和活性增强。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REDD 1对NRF 2的影响 糖尿病实验模型中的合成。2.描述REDD 1对Nrf 2降解的影响， 糖尿病的实验模型为了验证我的假设，我将进行一项实验， 从细胞培养到完整小鼠的模型系统，以及用于分析 mRNA翻译。该研究金还将提供两个关键的培训机会。首先，我将训练 博士Sui Wang（斯坦福大学）开发操纵视网膜基因表达所需的技能 in vivo.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， 发展技术专长，以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。关于成果，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变的发展起着重要作用。 确定这种机制是重要的，因为它将为制定 旨在解决糖尿病视网膜病变的分子基础的预防和/或治疗干预 促进健康视力。

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.