Molecular Mechanisms of corneal wound repair

角膜伤口修复的分子机制

• 批准号: 10470861
• 负责人: Vickery E Trinkaus-Randall
• 金额: $ 40.01万
• 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/10470861
• 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; 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

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.

据世界卫生组织称，2 型糖尿病是导致糖尿病的主要疾病之一 增加伤口愈合不当的风险，现在被认为是伤口愈合的主要原因之一 可预防的失明。受伤后，伤口边缘的相邻上皮细胞必须进行通信并 协调他们的反应以向前迈进。我们在该提案中的目标是检查相互作用是否 P2X7 和 pannexin1 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 pannexin1 介导 P2X7 诱导的通讯和细胞运动。这 2 种蛋白质对于离子至关重要 动员、ATP 运输和信号复合物在炎症和疼痛中发挥作用。其他 研究人员表明 P2X7 在 2 型糖尿病的发病机制中发挥作用，包括 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在角膜上皮中证明了： 1. P2X7 和 pannexin1 定位随损伤而变化，并在控制角膜前缘附近增强； 2. 抑制 P2X7 减少细胞之间的通讯，改变运动成分并改变酪氨酸 粘附斑和接头蛋白的磷酸化； 4. P2X7 和 pannexin1 在体外相互作用，我们 初步数据表明 P2X7 和 pannexin1 在对照小鼠中使用接近度相互作用 连接测定； 5. 我们有初步数据表明，未受伤者中的 pannexin1 差异极小 控制和糖尿病角膜上皮；然而，对伤害的反应有所不同； 6. 抑制 pannexin1 阻碍对照角膜和体外的细胞迁移。这些使我们假设 P2X7- pannexin1 在角膜上皮中的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，并需要调节细胞骨架和细胞运动所需的力。这 我们要解决的具体目标是：确定嘌呤受体关联的变化如何 pannexin1 在角膜伤口愈合过程中受到调节；确定这 2 种物质是否受到抑制和/或激活 蛋白质改变肌动蛋白细胞骨架和细胞运动的组织，并决​​定伤口是否愈合 运动性是由 pannexin-P2X7 相互作用施加的力介导的，如果它受到 基材刚度。 我们的数据表明治疗延迟角膜上皮的治疗方法可能会愈合， 以及 2 型糖尿病患者的复发性角膜糜烂。