Fascin Protrusions in ECM-Cytoskeletal Networking

ECM-细胞骨架网络中的肌成束蛋白突出

基本信息

批准号：
7072797
负责人：
JOSEPHINE C ADAMS
金额：
$ 29.13万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-06-01 至 2008-05-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Protrusive and contractile cell-matrix adhesions have important roles in cell adhesion, movement and signaling. Until recently, there has been a paucity of systems with which to research cell protrusions at molecular level. This laboratory previously discovered that thrombospondin-1, a cell-adhesion molecule, specifically induces protrusive lamellipodia, spikes and filopodia that contain the actin cross-linking protein, fascin, in core F-actin bundles. These fascin-containing structures are necessary for cell adhesion and migration on TSP-1 substrata, contribute to adhesion and migration on fibronectin substrata and are also formed in response to certain peptide growth factors. We have identified a novel molecular pathway by which thrombospondin-1 acts through clustering of the cell-surface proteoglycan syndecan-1 to stimulate the assembly of fascin-containing protrusions. The central hypothesis of the proposed research is that molecular components necessary for assembly of fascin-containing cell protrusions participate in protein complexes that contribute to the regulation of cell-matrix adhesion and cell movement. The proposed research focuses on the elucidatation of the molecular processes downstream of syndecan- 1 that mediate the organization of fascin cytoskeletal structures, the initial integration of these molecular processes into cell signaling networks, and the identification of mechanisms by which fascin-containing protrusions contribute to cell movements in three-dimensional matrix. These goals will be addressed experimentally by, 1), elucidation of the molecular requirements for regions within the syndecan-1 extracellular and cytoplasmic domains in transducing the organization of fascin protrusions; 2), the discovery of key binding partners for the V region of the syndecan-1 cytoplasmic domain and how these are integrated into regulation of fascin organization through the activation of Cdc42 and Rac small GTPases ; 3), the identification of molecular processes by which fascin structures contribute to migration in 3-dimensional matrix. Success in these complementary aims will advance new knowledge and understanding of the roles of fascin-based cytoskeletal structures in the important fundamental cellular activities of matrix-adhesion and cell migration. By opening up a new understanding of the molecular processes downstream of a specific extracellular inducer of fascin-containing protrusions, the proposed research will develop knowledge that is an essential basis for further investigation of fascin protrusions and associated regulatory molecules as therapeutic targets.

描述（由申请人提供）：突出和收缩的细胞基质粘附在细胞粘附、运动和信号传导中具有重要作用。直到最近，还缺乏在分子水平上研究细胞突起的系统。该实验室之前发现，血小板反应蛋白-1（一种细胞粘附分子）可特异性诱导突出的片状伪足、刺突和丝状伪足，这些伪足在核心 F-肌动蛋白束中含有肌动蛋白交联蛋白肌成束蛋白。这些含有肌成束蛋白的结构对于 TSP-1 基质上的细胞粘附和迁移是必需的，有助于纤连蛋白基质上的粘附和迁移，并且也是响应某些肽生长因子而形成的。我们已经确定了一种新的分子途径，血小板反应蛋白-1 通过细胞表面蛋白聚糖 syndecan-1 的聚集来发挥作用，刺激含有肌成束蛋白的突起的组装。该研究的中心假设是，组装含肌成束蛋白的细胞突起所需的分子成分参与了有助于调节细胞基质粘附和细胞运动的蛋白质复合物。拟议的研究重点是阐明 Syndecan-1 下游介导肌成束蛋白细胞骨架结构组织的分子过程，这些分子过程最初整合到细胞信号网络中，以及识别含肌成束蛋白的突起有助于形成肌成束蛋白的机制。三维矩阵中的细胞运动。这些目标将通过实验来解决，1) 阐明 syndecan-1 细胞外和细胞质域内区域在转导肌成束蛋白突起组织中的分子要求； 2)、发现 syndecan-1 胞质结构域 V 区的关键结合伴侣，以及如何通过激活 Cdc42 和 Rac 小 GTPase 将这些结合蛋白整合到肌成束蛋白组织的调节中； 3)，鉴定肌成束蛋白结构有助于在3维基质中迁移的分子过程。这些互补目标的成功将促进对基于肌成束蛋白的细胞骨架结构在基质粘附和细胞迁移的重要基本细胞活动中的作用的新认识和理解。通过开启对含肌成束蛋白突起的特定细胞外诱导剂下游分子过程的新认识，拟议的研究将开发知识，这些知识是进一步研究肌成束蛋白突起和相关调节分子作为治疗靶点的重要基础。