Role of Oxidative Stress in Pathogenesis of Fuchs Endothelial Corneal Dystrophy

氧化应激在福克斯内皮性角膜营养不良发病机制中的作用

• 批准号: 8957350
• 负责人: Ula V. Jurkunas
• 金额: $ 50.84万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8957350
• 关键词: Accounting; Agonist; Antioxidants; Apoptosis; Base Excision Repairs; Biogenesis; Blindness; Cell Aging; Cell Death; Cell Line; Cell Nucleus; Cells; Chronic; Cornea; Corneal Endothelium; Corneal dystrophy; Corneal edema; DNA Adduction; DNA Adducts; DNA Damage; DNA Repair; Data; Deposition; Descemet' s membrane; Development; Disease; Elderly; Endothelial Cells; Endothelium; Enzymes; Estrogen Metabolism; Estrogens; Etiology; Extracellular Matrix; Female; Grant; Human; In Vitro; Incidence; Investigation; Ion Pumps; Keratoplasty; Knockout Mice; Laboratories; Lead; Lesion; Light; Link; Mesenchymal; Mitochondria; Mitochondrial DNA; Modeling; Morphology; Mus; NADP; NF-E2-related factor 2; Nuclear; Oxidants; Oxidative Stress; Pathogenesis; Pathway interactions; Patients; Predisposition; Process; Pump; Quinones; Role; Severities; Signal Pathway; Telomerase; Tissues; UVA induced; Woman; age related; base; cytotoxic; enzyme pathway; genotoxicity; human old age (65+); in vivo; irradiation; male; mitochondrial dysfunction; oxidative DNA damage; prevent; public health relevance; repaired; response; ultraviolet

 DESCRIPTION (provided by applicant): Fuchs Endothelial Corneal Dystrophy (FECD), a common age-related dystrophy, which is more prevalent in women, is of unknown etiology. In FECD, corneal endothelial (CE) cell loss is accompanied by abnormal extracellular matrix (ECM) deposition in the form of guttae. Our laboratory was the first to link oxidative stress with FECD pathogenesis. Specifically, we showed that in FECD, oxidant-antioxidant imbalance due to suboptimal Nrf2-regulated antioxidant defense, including a decline in its transcriptional target NQO1, leads to oxidative DNA damage, mitochondrial dysfunction, and apoptosis. However, little is known regarding the mechanism of guttae formation and the predominance of FECD development in women. The mechanism of the observed DNA damage in cellular degeneration in FECD is not known. Building upon our previous findings, we propose to investigate if oxidant-antioxidant imbalance in FECD leads to endothelial mesenchymal transition (EMT) with concomitant ECM deposition, and if this imbalance causes the formation of cytotoxic estrogen metabolites, which are known to accumulate in tissues with the defective Nrf2-NQO1 and cause DNA damage, more commonly in women. Since our preliminary studies indicate a defective DNA damage response (DDR) in FECD, we will determine whether it leads to decreased DNA damage repair, cellular senescence, and apoptosis. Our proposal seeks to explore three different disease mechanisms: EMT, estrogen genotoxicity, and DDR, all of which either independently or collectively may lead to the development of FECD. Our study is significant, as the investigation of the mechanisms involved in the oxidative stress-induced cellular damage will provide new treatment targets for FECD. In order to achieve these aims, we will use our newly developed telomerase-immortalized human CE cells and the in vivo oxidative stress model based on the ultraviolet-A irradiation of mouse corneas. This model is specifically relevant to our study since it replicates in mice the CE morphological changes with guttae-like lesions and apoptosis, characteristically seen in FECD patients. Our Specific Aims are: Aim 1: Determine whether oxidant-antioxidant imbalance seen in FECD activates EMT that leads to aberrant ECM deposition seen in guttae. This aim is based on the hypothesis that in FECD chronic oxidative stress causes endothelial cells to undergo EMT and enhanced ECM deposition leading to guttae formation that can be modified by activation of Nrf2 pathway. Aim 2: Investigate the role of reactive estrogen metabolites in initiating an estrogen genotoxic pathway that may account for higher incidence and severity of FECD in women. This aim is based on the hypothesis that altered levels of estrogen-metabolizing enzymes lead to imbalanced estrogen metabolism and increased formation of depurinating estrogen-DNA adducts in FECD. Aim 3: Investigate the role of DDR on mitochondrial and nuclear DNA repair during oxidative stress- induced damage of CE in FECD. This aim is based on the hypothesis that impaired DDR leads to deficient DNA repair that results in mitochondrial dysfunction, cellular senescence, and apoptosis.

 描述（由适用提供）：富氏内皮角膜营养不良（FECD）是一种常见的年龄相关性营养不良，在女性中更为普遍，是未知的病因。在FECD中，角膜内皮（CE）细胞损失伴有异常的细胞外基质（ECM）沉积，形式为guttae。我们的实验室是第一个将氧化应激与FECD发病机理联系起来的实验室。具体而言，我们表明，在FECD中，氧化剂 - 抗氧化剂失衡是由于次优NRF2调节的抗氧化剂防御，包括其转录靶标的下降 NQO1，导致氧化物DNA损伤，线粒体功能障碍和凋亡。然而，关于牙齿形成的机理和女性FECD发展的占主导地位，知之甚少。 FECD中观察到的DNA变性中观察到的DNA损伤的机制尚不清楚。在我们先前的发现的基础上，我们建议研究FECD中的氧化物抗氧化剂是否会导致内皮间质质过渡（EMT）和伴随的ECM沉积，并且这种失衡会导致细胞毒性雌激素的代谢物的形成，而这些雌激素的代谢物已知会损害妇女的较常见nrf2-nrf2-nrf2-NQO，并构成较常见的妇女。由于我们的初步研究表明FECD中的DNA损伤反应（DDR）有缺陷，因此我们将确定它是否导致DNA损伤修复，细胞感​​应和凋亡。我们的建议旨在探索三种不同的疾病机制：EMT，雌激素遗传毒性和DDR，所有这些机制都独立或集体地可能导致FECD的发展。我们的研究很重要，因为氧化应激诱导的细胞损伤中涉及的机制的投资将为FECD提供新的治疗靶标。为了实现这些目标，我们将使用新开发的端粒酶增压人CE细胞和基于紫外线的体内氧化物应激模型 - 小鼠角膜的辐射。该模型与我们的研究特别相关，因为它在小鼠中复制了CE形态学的变化，而类似肠内的病变和细胞凋亡，在FECD患者中特征是。我们的具体目的是：目标1：确定在FECD中看到的氧化剂 - 抗氧化剂失衡是否激活EMT，这会导致Guttae中看到的异常ECM沉积。该目的基于以下假设：在FECD慢性氧化物中，会导致内皮细胞经历EMT并增强ECM沉积，从而导致Guttae形成，从而可以通过激活NRF2途径来改变。 AIM 2：研究反应性雌激素代谢产物在启动雌激素遗传毒性途径中的作用，该途径可能占女性的发病率和严重程度。该目的基于以下假设：雌激素代谢酶的水平改变导致雌激素代谢不平衡，并增加了fecd中雌激素-DNA加合物的形成。 AIM 3：研究DDR在氧化物胁迫诱导CE在FECD中损害期间线粒体和核DNA修复的作用。该目的基于以下假设：DDR受损会导致DNA修复不足，从而导致线粒体功能障碍，细胞感应和凋亡。