MECHANISM OF FIBRINOGEN ASSEMBLY IN EXTRACELLULAR MATRIX

损伤后的血管修复过程是在一个 纤维蛋白原动态相互作用的时空方式 (FBG)，纤维蛋白和细胞外基质（ECM）与细胞的 血管壁这项建议的长远目标是 描述了血管形成的细胞和分子机制， 修复发生。蛋白聚糖是受体的基本组成部分- 生长因子相互作用、细胞-细胞识别系统和细胞-ECM 粘附过程相互作用协调刺激细胞 细胞修复所需的增殖和迁移。因为蜂窝小区 表面硫酸乙酰肝素蛋白聚糖在 这些过程，以及肝素的广泛临床应用 在预防血栓形成方面， 肝素与纤维蛋白（原）的结合具有重要意义， 了解肝素调节止血的机制， 纤维蛋白溶解、血管生成和组织重塑。初步研究 证明FBG，而不是纤维蛋白，被纳入ECM，导致 隐藏的肝素结合域（HBD）的暴露。我们将测试 假设FBG通过其HBD在肝素中起积极作用， 以及硫酸肝素对细胞-细胞和细胞-基质相互作用的调节 参与船舶维修。拟议的实验将阐明 将FBG组装成预制的成熟基质的机制， 极化和间质细胞类型，以确定功能作用 基质-FBG介导的信号转导， 细胞修复和组织重塑的有序过程。具体目标1 这一建议将确定基本的结构领域， 基质FBG，并确定相互细胞表面受体， 基质成分支持FBG组装到ECM中。的 细胞生物学、蛋白质生物化学、共聚焦扫描技术 激光细胞术、荧光显微镜和免疫检测将 用于表征组装的关键配体和受体 将FBG导入ECM。具体目标2将是检查细胞 对沉积并组装成成熟ECM的FBG的响应。调制 细胞增殖和迁移的FBG沉积到ECM，包括 所涉及的信号转导途径，将使用细胞 和分子生物学技术。定义结构/功能 FBG、纤维蛋白和ECM与细胞相互作用的关系 将提供一个新的理解宿主的反应 涉及血栓形成治疗和治疗性损伤 操纵血管生成。

The process of blood vessel repair following injury is carried out in a temporal and spatial manner by the dynamic interaction of fibrinogen (FBG), fibrin and the extracellular matrix (ECM) with cells of the vessel walls. The long term objectives of this proposal are to characterize the cellular and molecular mechanisms by which vessel repair occurs. Proteoglycans are essential components of the receptor- growth factor interactions, cell-cell recognition systems, and cell-ECM adhesion processes that interact coordinately to stimulate cell proliferation and migration required for cellular repair. Because cell surface heparan sulfate proteoglycans perform essential functions in these processes, and with the extensive clinical application of heparin in the prevention in the prevention of thrombosis, the potential for heparin binding to fibrin(ogen) has important implications in understanding the mechanisms of heparin modulation of hemostasis, fibrinolysis, angiogenesis and tissue remodeling. Preliminary studies demonstrate that FBG, not fibrin, is incorporated into ECM, resulting in exposure of a cryptic heparin binding domain (HBD). We will test the hypothesis that FBG, through its HBD, plays an active role in heparin and heparin sulfate modulation of cell-cell and cell-matrix interactions involved in vessel repair. The proposed experiments will elucidate the mechanisms by which FBG is assembled into preformed, mature matrices of polarized and interstitial cell types to determine the functional role of matrix-FBG in mediating signal transduction to bring about the ordered process of cell repair and tissue remodeling. Specific Aim 1 of this proposal will be to define the essential structural domains of matrix FBG, and to determine the reciprocal cell-surface receptors and matrix constituents that support assembly of FBG into ECM. The techniques of cell biology, protein biochemistry, confocal scanning laser cytometry, fluorescence microscopy, and immunodetection will be used to characterize the ligands and receptors critical for the assembly of FBG into ECM. Specific Aim 2 will be to examine the cellular responses to FBG deposited and assembled into mature ECM. Modulation of cell proliferation and migration by FBG deposited into ECM, including the signal transduction pathways involved, will be examined using cell and molecular biology techniques. Defining the structure/function relationships involved in FBG, fibrin and the ECM interaction with cells of the vessel wall will provide a new understanding of the host response to injury with implications for treatment of thrombosis and therapeutic manipulation of angiogenesis.