Targeting the Etiology of Diabetic Retinopathy

针对糖尿病视网膜病变的病因

• 批准号: 10270082
• 负责人: Michael D. Dennis
• 金额: $ 8.27万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10270082
• 关键词: Address; Adult; Affect; Age; American; Antioxidants; Automobile Driving; Binding; Biological Models; Blindness; C-terminal; Cell Culture Techniques; Cells; Centers for Disease Control and Prevention (U.S.); Clinical Trials; Complement; Complication; Contrast Sensitivity; DNA Damage; Developed Countries; Development; Diabetes Mellitus; Diabetic Retinopathy; Diabetic mouse; Disease Progression; Early Intervention; Edema; Electroretinography; Etiology; Event; Exposure to; Functional disorder; Gene Expression; Genetic Transcription; Grant; Half-Life; Human; Hyperglycemia; Impairment; Individual; Insulin-Dependent Diabetes Mellitus; Laboratories; Light; Mediating; Messenger RNA; Microvascular Dysfunction; Molecular; Muller' s cell; Mus; Non-Insulin-Dependent Diabetes Mellitus; Onset of illness; Optical Coherence Tomography; Oxidative Stress; Parents; Pathology; Patients; Prediabetes syndrome; Proteins; Protocols documentation; Reading; Research; Retina; Rodent Model; Role; S-nitro-N-acetylpenicillamine; Small Interfering RNA; Streptozocin; TXN gene; TXNIP gene; Technical Expertise; Techniques; Testing; Therapeutic; Ubiquitination; Vascular Endothelial Growth Factors; Vision; Visual; Visual Acuity; Visual Fields; aged; bevacizumab; biological adaptation to stress; clinically significant; design; diabetic; diabetic patient; doctoral student; improved; in vivo; knock-down; macular edema; multicatalytic endopeptidase complex; neovascularization; novel therapeutics; pre-clinical; pre-doctoral; prevent; protein degradation; protein expression; skills; training opportunity; virtual; visual dysfunction

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 project proposed for this research supplement to enhance diversity is designed to complement the aims of the parent R01 by examining the impact of diabetes on degradation of the stress response protein REDD1 (regulated in development and DNA damage 1) in the retina. Our laboratory has demonstrated that REDD1 expression is increased in the retina of diabetic mice, and is necessary for diabetes-induced retinal pathology and functional deficits in vision. Moreover, siRNA-mediated REDD1 knockdown improves best-corrected visual acuity in patients with diabetic macular edema. Thus, identification of the molecular events responsible for diabetes-induced REDD1 expression in the retina is of high clinical significance. In studies conducted as part of the Parental Grant, we recently found evidence that a post-transcriptional mechanism was potentially responsible for the increase in REDD1 protein in the retina of diabetic mice. Specifically, in the retina of streptozotocin-induced diabetic mice, REDD1 protein expression was increased in the absence of a change in REDD1 mRNA abundance. The central hypothesis of this supplement is that diabetes-induced oxidative stress promotes REDD1 interaction with TXNIP (thioredoxin interacting protein) in a manner that prevents the ubiquitination and subsequent proteasomal degradation of REDD1. To test the hypothesis, the applicant will pursue an experimental protocol involving model systems ranging from intact mice to retinal cell culture. Aim 1 will use cutting-edge molecular techniques (e.g. SNAP- tagging) to evaluate the impact of diabetes on retinal REDD1 protein degradation. Aim 2 will examine the impact of TXNIP on retinal REDD1 protein turnover in the context of diabetes. The specific mechanism(s) responsible for the increase in REDD1 that is associated with the diabetes-induced decline in visual function has not been previously established, and thus represents a significant gap in our understanding of the molecular events that cause visual dysfunction in diabetic retinopathy. Overall, the studies supported by this supplement will greatly enhance the parent R01 by exploring why hyperglycemic conditions promote retinal REDD1 protein content. The proposed project represents a focused thesis project that provides two key training opportunities for a promising pre-doctoral student. First, the applicant will develop the technical expertise to assess the impact of diabetes on retinal pathophysiology using optical coherence tomography, electroretinograms, and virtual optomotry. The applicant will also develop skills necessary to manipulate retinal gene expression in vivo using AAV.

项目总结 糖尿病视网膜病变是美国工作年龄失明的主要原因，影响超过三分之一的 约2000万糖尿病患者。本项目建议对本研究进行补充，以增强 多样性旨在通过检查糖尿病对以下方面的影响来补充父母R01的目标 应激反应蛋白Redd1(在发育和DNA损伤1中调节)的降解 视网膜。我们的实验室已经证明，Redd1在糖尿病小鼠的视网膜中表达增加，并且 是糖尿病引起的视网膜病理和视力功能障碍所必需的。此外，siRNA介导的 Redd1基因敲除可改善糖尿病黄斑水肿患者的最佳矫正视力。因此， 糖尿病诱导视网膜中Redd1表达的分子事件的鉴定 具有较高的临床意义。在作为父母助学金一部分进行的研究中，我们最近发现有证据表明， 转录后机制可能是视网膜Redd1蛋白增加的原因 糖尿病小鼠。特别是，在链脲佐菌素诱导的糖尿病小鼠的视网膜中，Redd1蛋白表达 在Redd1mRNA丰度没有变化的情况下，表达增加。这一点的中心假设是 补充说，糖尿病引起的氧化应激促进Redd1与TXNIP(硫氧还蛋白)的相互作用 相互作用蛋白)，以防止泛素化和随后的蛋白酶体降解 红色1。为了验证这一假设，申请者将进行一项涉及模型系统的实验方案 从完整的小鼠到视网膜细胞培养。AIM 1将使用尖端分子技术(例如SNAP- 标签)，以评估糖尿病对视网膜Redd1蛋白降解的影响。Aim 2将研究 糖尿病背景下TXNIP对视网膜Redd1蛋白周转的影响。具体机制(S) 导致与糖尿病引起的视觉功能下降有关的Redd1的增加 并不是以前确定的，因此在我们对 糖尿病视网膜病变中导致视觉功能障碍的分子事件。总体而言，该项目支持的研究 补充剂将通过探索为什么高血糖条件促进视网膜来极大地增强父母R01 Redd1蛋白质含量。建议的项目代表了一个有重点的论文项目，它提供了两个关键 为一名有前途的博士生提供培训机会。首先，申请者将制定技术 使用光学相干断层扫描评估糖尿病对视网膜病理生理学的影响的专业知识， 视网膜电信号和虚拟验光。申请者还将发展操作视网膜所需的技能。 利用AAV进行体内基因表达。