Effects of systemic disease on corneal epithelial pathophysiology

全身性疾病对角膜上皮病理生理学的影响

• 批准号: 10468242
• 负责人: DANIELLE M. ROBERTSON
• 金额: $ 51.8万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468242
• 关键词: Affect; Apoptosis; Biogenesis; Blindness; Cell Nucleus; Cell Survival; Cells; Complications of Diabetes Mellitus; Cornea; Corneal Diseases; Corneal dystrophy; Data; Diabetes Mellitus; Diabetic mouse; Disease; Dose; Economic Burden; Epidemic; Epithelial Cell Proliferation; Epithelial Cells; Family; Functional disorder; Future; Generations; Goals; Grant; Health; Homeostasis; Human; Hyperglycemia; In Vitro; Insulin; Insulin Receptor; Insulin-Like Growth Factor Binding Protein 3; Insulin-Like Growth Factor I; Insulin-Like-Growth Factor I Receptor; Laboratories; Lead; Ligands; Maintenance; Mediating; Mediator of activation protein; Metabolic; Metabolism; Mitochondria; Mitochondrial DNA; Molecular; Morbidity - disease rate; Mus; Nerve; Nerve Plexus; Nuclear; Outcome; Pain; Pathway interactions; Patients; Process; Production; Quality Control; Quality of life; Regulation; Regulatory Pathway; Reporting; Respiration; Risk; Role; Signal Transduction; Small Interfering RNA; Somatomedins; Streptozocin; Stress; System; Systemic disease; Testing; Time; Translating; United States; Visual impairment; Voltage-Dependent Anion Channel; base; clinically significant; compliance behavior; corneal epithelial wound healing; corneal epithelium; cost; diabetic; environmental change; glucose uptake; in vivo; insulin secretion; knock-down; loss of function; mitochondrial dysfunction; molecular array; mouse model; new therapeutic target; novel; novel therapeutics; prevent; receptor; response; secretory protein; traditional therapy; trafficking; translational impact; type I diabetic

PROJECT SUMMARY Corneal complications from diabetes are common, often very painful, can negatively impact quality of life, and lead to permanent visual impairment. Owing to the vast array of molecular pathways that are altered in the diabetic cornea, traditional therapies are often not sufficient, due to abnormal cellular responses and the loss of trophic support from corneal nerves. Our prior studies have shown that the insulin-like growth factor (IGF) system is altered in diabetes. This includes a significant increase in the pleiotropic secretory protein, IGF-binding protein-3 (IGFBP-3) in diabetic human tears that correlates with loss of the corneal subbasal nerve plexus. While corneal epithelial cells do not require insulin for glucose uptake, our laboratory has found instead that insulin has an essential role in mediating expression of the IGF type 1 receptor (IGF-1R), insulin receptor (INSR), and IGFBP-3. Further interrogation of these pathways led to the generation of novel and exciting findings in corneal epithelial cells (CECs) that form the basis for the current proposal. Of high relevance to this proposal, all of our findings implicate the IGF-1 system as a mediator of mitochondrial function and stability. This is clinically significant since mitochondrial damage is a major underlying cause of disease morbidity in diabetes and other systemic diseases. Based on these findings, we propose to test two central hypotheses: (1) that insulin and IGFBP-3 regulate mitochondrial quality control and stability by coordinating mitochondrial-nuclear crosstalk in response to stress; and, (2) that interactions between the voltage dependent anion channel VDAC1, IGF-1R, and INSR mediate mitochondrial biogenesis and stability. We will test these hypotheses in the following aims. Aim 1. Compare the effects of IGF-related ligands, receptors, and IGFBP-3 on signaling, proliferation, and metabolism in normoglycemic versus hyperglycemic CEC culture in vitro and the diabetic mouse corneal epithelium in vivo. Aim 2: Compare the effects of IGF-related ligands, receptors, and IGFBP-3 on mitochondrial biogenesis, stability through interactions with VDAC1, and dynamics (fission/fusion) in CECs in vitro and the diabetic mouse corneal epithelium in vivo. Aim 3: Compare the effects of IGF-related ligands, receptors, and IGFBP-3 on mitophagy and apoptosis in CECs in vitro and the diabetic mouse corneal epithelium in vivo. Mitochondria are well known to regulate signals that drive energy production and cell survival, and their dysfunction has been implicated in a wide range of diseases. These studies will provide the first comprehensive picture detailing the role of the IGF family in mediating mitochondrial stability and function in the healthy cornea and in the pathobiology of disease. The outcomes could represent a major paradigm shift in our understanding and future treatment of the diabetic corneal epithelium and other corneal diseases and dystrophies where mitochondrial function is altered.

项目概要 糖尿病引起的角膜并发症很常见，通常非常痛苦，会对生活质量产生负面影响，并且 导致永久性视力障碍。由于大量的分子途径发生了改变 对于糖尿病性角膜，由于异常的细胞反应和角膜的丧失，传统疗法往往是不够的。 来自角膜神经的营养支持。 我们之前的研究表明，胰岛素样生长因子（IGF）系统在糖尿病中发生了改变。这 包括糖尿病患者中多效性分泌蛋白 IGF 结合蛋白 3 (IGFBP-3) 的显着增加 人类的眼泪与角膜基底下神经丛的丧失有关。虽然角膜上皮细胞不 需要胰岛素来摄取葡萄糖，我们的实验室发现胰岛素在 介导 IGF 1 型受体 (IGF-1R)、胰岛素受体 (INSR) 和 IGFBP-3 的表达。更远 对这些途径的研究导致了角膜上皮细胞中新颖且令人兴奋的发现的产生 （CEC）构成当前提案的基础。我们所有的发现都与该提案高度相关 表明 IGF-1 系统是线粒体功能和稳定性的调节剂。这具有临床意义 因为线粒体损伤是糖尿病和其他疾病发病的一个主要根本原因 全身性疾病。 基于这些发现，我们建议检验两个中心假设：(1) 胰岛素和 IGFBP-3 调节 通过协调线粒体-核串扰响应来控制线粒体质量和稳定性 强调； (2) 电压依赖性阴离子通道 VDAC1、IGF-1R 和 INSR 介导线粒体生物发生和稳定性。我们将在以下目标中测试这些假设。 目标 1. 比较 IGF 相关配体、受体和 IGFBP-3 对信号传导、增殖和生长的影响 正常血糖与高血糖 CEC 体外培养物和糖尿病小鼠角膜的代谢 体内的上皮细胞。目标 2：比较 IGF 相关配体、受体和 IGFBP-3 对线粒体的影响 体外 CEC 的生物发生、通过与 VDAC1 相互作用的稳定性以及动力学（裂变/融合） 糖尿病小鼠角膜上皮体内。目标 3：比较 IGF 相关配体、受体和 IGF 的作用 IGFBP-3 对体外 CEC 和体内糖尿病小鼠角膜上皮细胞线粒体自噬和细胞凋亡的影响。 众所周知，线粒体可以调节驱动能量产生和细胞存活的信号，其 功能障碍与多种疾病有关。这些研究将提供第一个 全面详细介绍了 IGF 家族在介导线粒体稳定性和功能中的作用 健康角膜和疾病的病理学。结果可能代表着重大的范式转变 在我们对糖尿病角膜上皮和其他角膜的理解和未来治疗中 线粒体功能改变的疾病和营养不良。