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Molecular Mechanisms of corneal wound repair

角膜伤口修复的分子机制

基本信息

批准号：
10674734
负责人：
Vickery E Trinkaus-Randall
金额：
$ 41.25万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10674734
关键词：
Actins Adaptor Signaling Protein Affect Agonist Area Basement membrane Biochemical Biological Biological Assay Blindness Blood-Retinal Barrier Cell Shape Cells Central Nervous System Communication Complex Cornea Corneal Injury Cytoskeletal Modeling Cytoskeleton Data Defect Diabetes Mellitus Diabetic mouse Diffuse Endocytosis Epithelial Cells Epithelium Event Eye Focal Adhesions Goals Health Homeostasis Impaired wound healing Impairment In Vitro Inflammation Injury Ions Ligation Mediating Mediator Medical Care Costs Methodology Microvascular Dysfunction Modeling Molecular Mus Non-Insulin-Dependent Diabetes Mellitus Organizational Change Pain Pathogenesis Pathology Persons Play Process Proteins Quality of life Reaction Recurrence Regulation Reporting Research Personnel Risk Role Signal Transduction Stains Surface Technology Therapeutic Tissues Traction Type 2 diabetic Tyrosine Phosphorylation Vision World Health Organization antagonist cell motility corneal epithelial wound healing corneal epithelium diabetic diabetic ulcer diabetic wound healing epithelial injury epithelial repair epithelial wound healing improved live cell imaging mechanotransduction member mouse model non-diabetic painful neuropathy protein degradation protein expression response response to injury sensor wound wound healing

项目摘要

According to the World Health Organization, type 2 diabetes is one of the leading pathologies that increases the risk of improper wound healing and is now considered to be one of the leading causes of preventable blindness. Upon injury, adjacent epithelial cells at the wound edge must communicate and coordinate their response to move forward. Our goal in this proposal is to examine if the interactions between P2X7 and pannexin1 are critical for homeostasis and wound repair. We have evidence that pannexin1 mediates P2X7-induced communication and cell motility. These 2 proteins are critical for ion mobilization, ATP transport and signaling complexes that play roles in inflammation and pain. Other investigators have shown that P2X7 has a role in the pathogenesis of type 2 diabetes including microvascular complications, impaired blood retinal barrier and neuropathic pain (Solini and Novak (2019). We and our collaborators have demonstrated in corneal epithelium that: 1. P2X7 and pannexin1 localization changes with injury and is enhanced near the leading edge of control corneas; 2. Inhibition of P2X7 diminishes communication between cells, alters components of motility and alters tyrosine phosphorylation of focal adhesion and adaptor proteins; 4. P2X7 and pannexin1 interact in vitro and we have preliminary data demonstrating the interaction of P2X7 and pannexin1 in control mice using proximity ligation assays; 5. We have preliminary data that there is minimal difference in pannexin1 in unwounded control and diabetic corneal epithelium; however the response to injury differs; and 6. Inhibition of pannexin1 impedes cell migration in control corneas and in vitro. These led us to hypothesize that P2X7- pannexin1 interaction in corneal epithelium is required for effective cell-cell communication and signaling in response to injury and is needed to regulate the cytoskeleton and forces required for cell motility. The specific aims that we will address are: determine how changes in association of purinoreceptors and pannexin1 are regulated during corneal wound healing; determine if inhibition and/or activation of these 2 proteins changes the organization of the actin cytoskeleton and cell motility and determine if wound healing and motility is mediated by a force exerted through the pannexin-P2X7 interaction and if it is affected by substrate stiffness. Our data suggest the potential for therapeutic approaches to treat delayed corneal epithelial would healing, and recurrent corneal erosions in type 2 diabetes.

根据世界卫生组织，2型糖尿病是主要的病理之一，增加了伤口愈合不良的风险，现在被认为是导致可预防的失明损伤后，伤口边缘的相邻上皮细胞必须沟通，协调他们的反应以向前推进。我们在这个提案中的目标是检查 P2 X7和pannexin 1之间的相互作用对于体内平衡和伤口修复至关重要。我们有证据表明 pannexin 1介导P2 X7诱导的通讯和细胞运动。这两种蛋白质对离子交换至关重要。在炎症和疼痛中发挥作用的细胞动员、ATP转运和信号复合物。其他研究人员已经表明，P2 X7在2型糖尿病的发病机制中起作用，微血管并发症、血视网膜屏障受损和神经性疼痛（Solini和Novak（2019）。我们和我们的合作者已经在角膜上皮中证明了：1。P2 X7和pannexin 1 定位随着损伤而改变，并且在对照角膜的前缘附近增强; 2.抑制 P2 X7减少细胞间的通讯，改变运动成分，改变酪氨酸粘着斑和衔接蛋白的磷酸化; 4. P2 X7和pannexin 1在体外相互作用，我们有初步的数据证明P2 X7和pannexin 1在对照小鼠中的相互作用，连接测定; 5.我们有初步的数据表明，在未受伤的人中，对照和糖尿病角膜上皮;然而，对损伤的反应不同;和6.抑制 pannexin 1在对照角膜和体外阻碍细胞迁移。这让我们假设P2 X7- 角膜上皮中的pannexin 1相互作用是有效的细胞间通讯和信号传导所必需的响应损伤，并且需要调节细胞运动所需的细胞骨架和力。的具体目标，我们将解决的是：确定如何变化的协会嘌呤受体和泛连接蛋白1在角膜伤口愈合期间受到调节;确定这2种蛋白的抑制和/或激活蛋白质改变了肌动蛋白细胞骨架的组织和细胞运动性，运动性是由通过pannexin-P2 X7相互作用施加的力介导的，如果它受到衬底刚度我们的数据表明，治疗延迟的角膜上皮愈合的治疗方法的潜力，和2型糖尿病中复发性角膜糜烂的关系。