FIBRONECTIN-MEDIATED CELL SIGNALING IN WOUND HEALING

伤口愈合中纤连蛋白介导的细胞信号传导

• 批准号: 6387242
• 负责人: SIOBHAN A CORBETT
• 金额: $ 25.05万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6387242
• 关键词: CHO cells; JUN kinase; biological signal transduction; cell cycle; cell growth regulation; cell migration; cell motility; cell proliferation; complementary DNA; cytoskeletal proteins; enzyme activity; extracellular matrix; fibrin; fibronectins; guanosinetriphosphatases; immunoprecipitation; integrins; intermolecular interaction; mitogen activated protein kinase; mutant; myosin light chain kinase; phosphorylation; platelet derived growth factor; protein structure function; wound healing

Following injury, the formation of a provisional extracellular matrix (ECM) from extravasated plasma proteins is a critical step in the wound repair process. The interactions between cell surface receptors and this "injury-associated" matrix provide important cues that can modulate the cellular response at the injury site leading to alterations in cell growth and gene expression. The long-term goal of this project is to define the molecular mechanisms of ECM-mediated signaling required for tissue repair. We hypothesize that fibronectin (FN), an essential adhesive component of the provisional ECM, is a key determinant of cell growth and survival during wound healing. We propose that FN modulates these important cellular behaviors through its interaction with the integrin receptor alpha5beta1. To determine the molecular mechanisms, both structural and biochemical, that are required for this process, we have developed an in vitro model that allows the incorporation of FN molecules into a three- dimensional (3D) fibrin matrix that mimics the provisional ECM found in vivo. We will use this system to test the hypotheses that: 1) the incorporation of FN into fibrin matrices enhances alpha5beta1-mediated matrix contraction by modulating cell motility and contractile force generation 2) alpha5beta1-FN interactions promote cell shape change and are essential for cell growth and survival in FN-fibrin matrices 3) the cytoplasmic domain of the alpha5 integrin subunit regulates FN-fibrin matrix contraction. The experiments outlined above are the first to specifically examine FN- mediated cell signaling in 3D culture. As such, they will provide important information regarding the determinants of cell behavior in an environment that closely approximates the early provisional wound matrix. This will improve our understanding of the fundamental processes of the injury response and contribute to the development of effective therapeutic strategies for acute and chronic injury.

受伤后，从外部血浆蛋白中形成临时细胞外基质（ECM）是伤口修复过程中的关键步骤。细胞表面受体与该“损伤相关”基质之间的相互作用提供了重要的线索，可以调节损伤部位的细胞反应，从而导致细胞生长和基因表达的改变。该项目的长期目标是定义组织修复所需的ECM介导的信号传导的分子机制。我们假设纤连蛋白（FN）是临时ECM的重要粘合剂成分，是伤口愈合过程中细胞生长和存活的关键决定因素。我们建议FN通过与整联蛋白受体α5BETA1的相互作用来调节这些重要的细胞行为。为了确定此过程所需的结构和生化的分子机制，我们开发了一种体外模型，该模型允许将FN分子掺入三维（3D）纤维蛋白基质中，该基质模拟了VIVO中发现的临时ECM。 We will use this system to test the hypotheses that: 1) the incorporation of FN into fibrin matrices enhances alpha5beta1-mediated matrix contraction by modulating cell motility and contractile force generation 2) alpha5beta1-FN interactions promote cell shape change and are essential for cell growth and survival in FN-fibrin matrices 3) the cytoplasmic domain of the alpha5 integrin subunit调节FN-纤维蛋白基质收缩。上面概述的实验是第一个专门检查3D培养中FN介导的细胞信号传导的研究。因此，他们将在紧密近似早期临时伤口矩阵的环境中提供有关细胞行为决定因素的重要信息。这将提高我们对伤害反应的基本过程的理解，并有助于发展有效的急性和慢性损伤治疗策略。