The role of endothelial connexins in vascular wound repair

内皮连接蛋白在血管伤口修复中的作用

基本信息

批准号：
10751730
负责人：
Meghan Sedovy
金额：
$ 4.25万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-10 至 2026-02-09
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751730
关键词：
Alanine Antibodies Aorta Area Arteries Biological Biology Biomedical Research Blood Vessels C-terminal Carotid Arteries Cell Communication Cell Culture Techniques Cell Line Cell Proliferation Cell Separation Cells Cessation of life Communication Confocal Microscopy Connexin 43 Connexins Coronary Thrombosis Coronary artery Data Data Analyses Defect Deoxyuridine Disease Endothelial Cells Endothelium Ensure Environment Equipment Failure Fellowship Female Gap Junctions Genetic Goals Health Heart Home Hour Human Immunofluorescence Immunologic Impaired healing Impairment Injections Injury Investigation Knock-out Knockout Mice Legal patent Life Ligation Link Measures Mediating Mentorship Microscope Mission Modeling Modernization Modification Mus Mutation Myocardial Infarction Operative Surgical Procedures Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Phosphorylation Inhibition Phosphorylation Site Phosphotransferases Plasmids Post-Translational Protein Processing Preparation Process Proliferating Proliferation Marker Proto-Oncogene Proteins c-akt Research Research Institute Research Personnel Role Saphenous Vein Serine Signal Pathway Signal Transduction Site Stents Survival Rate Technical Expertise Techniques Testing Therapeutic Thrombus Tissues Training Transfection Travel United States National Institutes of Health Universities Up-Regulation Vascular Diseases Vascular Endothelial Cell Vascular Graft Virginia Western Blotting Work animal facility burden of illness cell growth cell injury cell motility design endothelial regeneration endothelial repair experience experimental study healing improved in vivo in vivo Model injured male migration mimetics novel pharmacologic protein expression repaired response restenosis sex stroke risk therapeutic target transcriptome sequencing vascular injury wound closure wound healing

项目摘要

|| Project Summary . Impaired endothelial wound healing in blood vessels like the coronary artery and saphenous vein is a primary contributor to vascular stent failure and deadly coronary thrombosis. Identifying the mechanism of delayed endothelial repair is essential for improving vessel patency and patient survival rates. As such, this proposal focuses on identifying regulators of normal endothelial healing and understanding how these factors are dysregulated when healing is impaired. Previous studies have shown connexin 43 (Cx43) gap junctions may regulate wound healing. However, the specific role of connexin 43 in endothelial cell (EC) healing is unknown. Preliminary data in this proposal indicates that ligation-induced vascular injury promotes increases in the expression of EC Cx43 surrounding the damaged area in the aorta and carotid arteries in mice. Cx43-mediated gap junction intracellular communication is controlled by phosphorylation at the Cx43 C-terminus. My preliminary data demonstrate that specific Cx43 phosphorylation at its serine (s) 368, associated with gap junction closure, is present during the final stages of EC wound healing in vivo. Cx43-s368 also reduced the rate of wound closure in cultured human EC. These findings inform the hypothesis that Cx43 expression and channel functions are critical for EC wound healing in large arteries. The aims of this proposal are to define the role of connexin 43 in endothelial wound healing (Aim 1), investigate if a loss of Cx43 limits healing in vivo (Aim 1), and test if channel functions, regulated by posttranslational modifications, improve/delay vascular repair (Aim 2). This investigation will be completed using a novel mouse carotid EC injury survival surgery in mice I developed, which will allow for the assessment of Cx43 expression in carotid EC during the healing process. Both genetic and pharmacological strategies will be used to alter Cx43 expression and phosphorylation in mouse injury models and in cultured human EC, and the impact of these modifications on the rate and quality of EC healing will be quantified. Additionally, RNAseq approaches will be used to identify Cx43 gap junction-dependent signaling in the regenerating endothelium. These aims will be accomplished under the mentorship of researchers with extensive experience in vascular biology, Dr. Scott Johnstone, Dr. Robert Gourdie, and Dr. Brant Isakson, at Virginia Tech’s Fralin Biomedical Research Institute (FBRI). The FBRI is home to research equipment including but not limited to animal facilities, confocal microscopes, flow cytometers, and cell culture equipment. This state- of-the-art research environment will allow for a detailed and mechanistic investigation. Ultimately, this study will characterize a novel role for Cx43 in vascular EC. This project is designed to identify new potential therapeutic targets in vascular disease that will promote the mission of the NIH to enhance patient health and to promote longer life free of illness.

||项目摘要。冠状动脉和隐静脉等血管中的内皮伤口愈合受损是主要的血管支架衰竭和致命冠状动脉血栓形成的贡献者。识别延迟的机制内皮修复对于提高血管通畅和患者存活率至关重要。因此，这个建议专注于确定正常内皮愈合的调节因子，并了解这些因素的状况如何治愈受损时失调。先前的研究表明连接蛋白43（CX43）间隙连接可能调节伤口愈合。但是，连接蛋白43在内皮细胞（EC）愈合中的特定作用尚不清楚。该提案中的初步数据表明，结扎引起的血管损伤促进小鼠主动脉和颈动脉受损区域周围的EC CX43的表达。 CX43介导的间隙连接在CX43 C端的磷酸化控制。我的初步数据表明，在其丝氨酸368处的特定CX43磷酸化，与间隙连接闭合相关，在体内EC伤口愈合的最后阶段存在。 CX43-S368还降低了伤口闭合率在文化的人类EC中。这些发现告知了CX43表达和通道功能的假设对于大动脉的EC伤口愈合至关重要。该提议的目的是确定连接蛋白43在内皮伤口愈合（AIM 1），研究CX43的损失是否限制了体内愈合（AIM 1），并测试是否通道受翻译后修饰调节的功能改善/延迟血管修复（AIM 2）。这项投资将使用我开发的小鼠中的新型小鼠颈动脉EC损伤手术完成，这将允许为了评估康复过程中颈动脉EC中CX43表达的表达。遗传和药理策略将用于改变小鼠损伤模型中的CX43表达和磷酸化在文化的人类EC中，这些修饰对EC治疗的速度和质量的影响将是量化。此外，RNASEQ方法将用于识别CX43间隙连接依赖性信号传导再生内皮。这些目标将在研究人员的指导下实现在血管生物学方面的丰富经验，Scott Johnstone博士，Robert Gourdie博士和Brant Isakson博士，AT 弗吉尼亚理工大学的弗拉林生物医学研究所（FBRI）。 FBRI是研究设备的家园但不限于动物设施，共聚焦显微镜，流式细胞仪和细胞培养设备。这个国家 - 艺术研究环境将允许进行详细的机械投资。最终，这项研究将表征CX43在血管EC中的新作用。该项目旨在确定新的潜在疗法血管疾病的靶标将促进NIH的使命，以增强患者健康并促进长寿没有疾病。