Molecular Mechanisms of corneal wound repair

角膜伤口修复的分子机制

• 批准号: 10296958
• 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
• 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/10296958
• 关键词: Actins; Adaptor Signaling Protein; Address; Affect; Agonist; Area; Basement membrane; Biochemical; Biological; Biological Assay; Blindness; Blood-Retinal Barrier; Cell Shape; Cells; Communication; Complex; Cornea; Corneal Injury; Cytoskeletal Modeling; Cytoskeleton; Data; Defect; Diabetes Mellitus; Diabetic mouse; Diffuse; Endocytosis; Epithelial; Epithelial Cells; Event; Eye; Focal Adhesions; Goals; Health; Homeostasis; Impaired wound healing; Impairment; In Vitro; Inflammation; Injury; Ions; Ligation; Mediating; Mediator of activation protein; Medical Care Costs; Methodology; Microvascular Dysfunction; Modeling; Molecular; Mus; Neuraxis; 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; Technology; Therapeutic; Tissues; Traction; Type 2 diabetic; Tyrosine Phosphorylation; Vision; World Health Organization; 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

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.

根据世界卫生组织的数据，2型糖尿病是 增加伤口愈合不当的风险，现在被认为是 可预防的失明。受伤后，在伤口边缘处的相邻上皮细胞必须通信，并且 协调他们向前发展的反应。我们在此提案中的目标是检查互动是否 P2X7和Pannexin1之间对于稳态和伤口修复至关重要。我们有证据表明 Pannexin1介导P2X7诱导的通信和细胞运动。这两种蛋白对于离子至关重要 动员，ATP传输和信号传导复合物在炎症和疼痛中起着作用。其他 研究人员表明，P2X7在2型糖尿病的发病机理中起作用 微血管并发症，血液视网膜屏障和神经性疼痛受损（Solini and Novak（2019）。 我们和我们的合作者在角膜上皮中证明了：1。P2X7和Pannexin1 损伤的定位变化，并在控制角膜的前缘附近得到增强。 2。抑制 P2X7减少了细胞之间的通信，改变运动的组成部分与改变酪氨酸 局灶性粘附和衔接蛋白的磷酸化； 4。P2X7和PANNEXIN1在体外相互作用，我们 有初步数据证明使用接近性P2X7和Pannexin1在对照小鼠中的相互作用 连接测定； 5。我们有初步数据，即未来的pannexin1差异很小 对照和糖尿病角膜上皮；但是，对伤害的反应有所不同。和6。抑制 Pannexin1阻碍了对照角膜和体外的细胞迁移。这些使我们假设P2X7- 有效的细胞 - 细胞通信和信号传导需要角膜上皮中的Pannexin1相互作用 响应损伤，需要调节细胞运动所需的细胞骨架和力。这 我们将要解决的具体目标是：确定嘌呤感受器和 在角膜伤口愈合过程中，Pannexin1受调节；确定这2的抑制和/或激活是否 蛋白质改变肌动蛋白细胞骨架和细胞运动的组织，并确定伤口愈合是否 运动性是由通过pannexin-p2x7相互作用施加的力介导的 底物刚度。 我们的数据表明，治疗延迟角膜上皮的治疗方法的潜力将愈合， 和2型糖尿病中的复发角膜侵蚀。