Dual Oxidase 2 as a New Target for Treatment of Diabetic Nephropathy

项目概要/摘要 肾小球上皮细胞/足细胞损伤是糖尿病肾病的突出病理特征 （DN）。在足细胞中，高血糖会导致裂隙隔膜蛋白、足突的改变 消失、细胞凋亡和细胞脱离，这些事件最终导致肾功能丧失。高 葡萄糖浓度（HG）和氧化应激是肾小球损伤的潜在介质 糖尿病。我们有证据表明双氧化酶 2 (Duox2) 在肾小球细胞中表达并且 有助于 HG 诱导的活性氧 (ROS) 产生以及足细胞损伤。 此外，我们还证明从 DuoxA-/- 小鼠中分离的肾小球受到保护 高葡萄糖诱导过氧化氢的产生。该提案的中心假设是 Duox2产生的ROS在糖尿病肾小球病变和足细胞损伤中发挥关键作用 肾。该提案的目标是利用体外和体内方法来建立 Duox2 在糖尿病环境中足细胞损伤中的重要性，并确定影响因素 调节 Duox2 活性/表达。 Duox 激活剂 (DuoxA) 的作用、翻译机制、 将探讨钙在 Duox2 激活中的作用。对于体内研究，我们将利用之前 收集了 DuoxA 功能受损的糖尿病和非糖尿病小鼠的肾皮质。由于没有 与足细胞内 Duox 酶表达和功能相关的数据可用，实验 提议应作为概念证明来证明针对 Duox/DuoxA 的实用性 系统以减少糖尿病介导的肾小球病变。有害行为的特征 Duox2 及其调节因子的鉴定将有助于新型治疗干预措施的设计 并将有助于建立治疗 DN 的辅助疗法。研究将强调相关性 开发肾脏中 Duox2 表达和活性的调节剂作为治疗剂 预防或逆转 DN。此外，该提案将加强研究和教育 化身大学的基础设施，将生物医学研究经验引入 少数族裔学生代表性不足，否则他们将缺乏这样的机会。

Project Summary/Abstract Glomerular epithelial cell/podocyte injury is a prominent pathological feature of diabetic nephropathy (DN). In podocytes, hyperglycemia causes alteration of slit diaphragm proteins, foot process effacement, apoptosis and cell detachment, events that ultimately results in loss of renal function. High glucose concentrations (HG) and oxidative stress are potential mediators of glomerular injury in diabetes. We have evidence that Dual oxidase 2 (Duox2) is expressed in glomerular cells and contributes to HG-induced reactive oxygen species (ROS) generation as well as podocyte injury. Additionally, we have demonstrated that glomeruli isolated from DuoxA-/- mice were protected from high glucose-induced hydrogen peroxide generation. The central hypothesis of this proposal is that the ROS generated by Duox2 play a pivotal role in glomerular lesions and podocyte injury in the diabetic kidney. The goals of this proposal are to utilize in vitro and in vivo approaches to establish the importance of Duox2 in podocyte injury in the diabetic environment and identify the factors that are modulating Duox2 activity/expression. The role of Duox activators (DuoxA), translational mechanisms, and calcium in Duox2 activation will be explored. For the in vivo studies, we will utilize previously collected kidney cortices from diabetic and non-diabetic mice where DuoxA function is impaired. As no data related to Duox enzyme expression and function within podocytes are available, the experiments proposed should serve as proof of concept to demonstrate the utility of targeting the Duox/DuoxA system to reduce diabetes-mediated glomerular lesions. Characterization of the deleterious actions of Duox2 and identification of its regulators will contribute to the design of novel therapeutic interventions and will help establish adjunct therapy to treat DN. The studies will stress the relevance of the development of modulators of Duox2 expression and activity in the kidney as therapeutic agents to prevent or reverse DN. Furthermore, this proposal will enhance the research and educational infrastructure at the University of the Incarnate Word, introducing biomedical research experiences to underrepresented minority students who would otherwise lack such opportunities.