Roles of endothelial tensins

内皮张力蛋白的作用

基本信息

批准号：
9419176
负责人：
SU HAO LO
金额：
$ 39.25万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2021-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9419176
关键词：
Actins Binding Biochemistry Biological Blindness Blood Vessels Cell Adhesion Cell Culture Techniques Cell Proliferation Cells Cellular biology Clinical Complex Confocal Microscopy Cues Cultured Cells Cytoplasmic Tail Cytoskeleton Development Differentiated Gene Disease Endothelial Cells Environment Extracellular Matrix Family Family member Fluorescence Microscopy Focal Adhesions Gene Expression Genetic Goals Growth and Development function Impairment In Vitro Integrins Knockout Mice Lead Link Malignant Neoplasms Mediating Microfilaments Molecular Molecular Biology Mus N-terminal PTB Domain Pattern Phenotype Phosphotyrosine Play Process Proteins Reporting Research Design Rheumatoid Arthritis Role Signal Pathway Signal Transduction Site Structure Subcellular structure System Testing Therapeutic Tissues Tube Tyrosine Wound Healing angiogenesis cell motility cell type diabetic extracellular in vivo innovation insight interdisciplinary approach member migration mouse model novel receptor response spatiotemporal tensin

项目摘要

Abstract The goal of the proposed studies is to understand the signaling transduction mechanism governing angiogenesis, an important process in growth and development of tissues, as well as in wound healing processes. It also occurs in diseases, such as cancer, diabetic blindness, and rheumatoid arthritis. In this project, we focus on the roles and mechanisms of tensin signaling in endothelial cells during angiogenesis. The tensin family plays critical roles in organizing the subcellular structure and mediating signaling transductions at focal adhesions, which are the transmembrane structures linking the extracellular matrix to the cytoskeleton. The four members of tensin (tensin1, tensin2, tensin3, and cten) bind to the cytoplasmic tails of  integrin through their PTB (phosphotyrosine-binding) domains and interact with actin filaments (except cten) via their N-terminal regions, allowing tensins to bridge the actin cytoskeleton to integrin receptors. In addition, tensins contain an SH2 (Src homology 2) domain that interacts with tyrosine-phosphorylated as well as non- phosphorylated proteins and form signaling complexes at focal adhesions. Our recent studies using knockout mice showed that lack of tensin1 impairs tube formation activities in endothelial cells and angiogenic processes in mice, indicating critical involvements of tensins in angiogenesis. However, not all tensins play similar roles in cellular activities. We found that tensin1 and tensin2 promote endothelial cell migration, a critical step during angiogenesis, whereas tensin3 suppresses it. Why highly homologous tensins exert opposite biological activities? By using fluorescent-tagged tensins and live-cell confocal microscopy, we observed that tensins show different spatiotemporal localization patterns in migrating cells. These findings lead us to investigate the roles and regulatory mechanisms of tensins in angiogenesis. We hypothesize that tensins regulate angiogenesis through their common and unique roles, which are dictated by their spatiotemporal localizations and associated molecules, in endothelial cell adhesion, migration, and vascular lumen formation. Three specific aims are proposed to (Aim 1) determine the primary control of spatiotemporal localizations of tensins during in vitro tube formation; (Aim 2) establish the roles and mechanisms of tensins in regulating endothelial cell tube formation; and (Aim 3) investigate the functions and regulatory mechanisms of tensins in angiogenesis using knockout mice. Our research design is innovative because it probes novel and distinct functions of tensins in endothelial cells, and employs a multidisciplinary approach that integrates biochemistry, cell and molecular biology, live-cell fluorescence microscopy, cell culture and mouse models to understand the roles of tensins in angiogenesis. This project has very high clinical and translational relevance that may offer new insights for therapeutic applications to angiogenic related diseases.

抽象的拟议研究的目标是了解控制信号转导机制血管生成，组织生长和发育以及伤口愈合的重要过程它也发生在癌症、糖尿病性失明和类风湿性关节炎等疾病中。项目中，我们重点研究血管生成过程中内皮细胞中张力蛋白信号传导的作用和机制。张力蛋白家族在组织亚细胞结构和介导信号转导方面发挥着关键作用粘着斑，是将细胞外基质与细胞骨架连接起来的跨膜结构。张力蛋白的四个成员（tensin1、tensin2、tensin3 和 cten）与  整合素的细胞质尾部结合通过其 PTB（磷酸酪氨酸结合）结构域并通过其与肌动蛋白丝（cten 除外）相互作用 N 末端区域，允许张力蛋白将肌动蛋白细胞骨架与整合素受体桥接。包含一个 SH2（Src 同源 2）结构域，可与酪氨酸磷酸化以及非磷酸化相互作用我们最近的研究使用敲除技术在粘着斑处磷酸化蛋白质并形成信号复合物。小鼠表明，缺乏tensin1会损害内皮细胞的管形成活性和血管生成过程在小鼠中，表明张力蛋白在血管生成中发挥着关键作用。然而，并非所有张力蛋白都发挥相似的作用。我们发现张力蛋白 1 和张力蛋白 2 促进内皮细胞迁移，这是细胞迁移过程中的关键步骤。为什么高度同源的张力蛋白发挥相反的生物学作用？通过使用荧光标记的张力蛋白和活细胞共聚焦显微镜，我们观察到张力蛋白显示迁移细胞中不同的时空定位模式。张力蛋白在血管生成中的作用和调节机制。通过它们的共同和独特的作用来促进血管生成，这是由它们的时空定位决定的以及相关分子在内皮细胞粘附、迁移和血管腔形成中的作用。提出了具体目标（目标 1）确定张力蛋白时空定位的主要控制（目标 2）建立张力蛋白在调节内皮细胞中的作用和机制细胞管形成；和（目标 3）研究张力蛋白的功能和调节机制我们的研究设计是创新的，因为它探索了新颖且独特的方法。内皮细胞中张力蛋白的功能，并采用整合生物化学的多学科方法，细胞和分子生物学、活细胞荧光显微镜、细胞培养和小鼠模型，以了解张力蛋白在血管生成中的作用该项目具有非常高的临床和转化相关性，可能会提供帮助。血管生成相关疾病的治疗应用的新见解。