Influence of extracellular matrix fibronectin on platelet derived growth factor signaling

细胞外基质纤连蛋白对血小板衍生生长因子信号传导的影响

• 批准号: 9121935
• 负责人: Christopher Farrar
• 金额: $ 2.91万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9121935
• 关键词: Animal Tarsus; Atherosclerosis; Attenuated; Binding; Biological; Calcium; Cells; Chimeric Proteins; Clinical; Controlled Study; Data; Deposition; Development; Diabetic ulcer; Disease; Down-Regulation; EGF gene; Embryo; Engineering; Extracellular Matrix; Extracellular Matrix Degradation; Extracellular Matrix Proteins; Fibroblast Growth Factor; Fibroblasts; Fibronectins; Fibrosis; Fluo 4; Functional disorder; Goals; Granulation Tissue; Healed; Homeostasis; Human; Intracellular Signaling Proteins; Knockout Mice; Laboratories; Libraries; Lung; Malignant Neoplasms; Measures; Mediating; Mesenchymal; Methods; Mitogens; Mus; Myofibroblast; PRKCA gene; Pathway interactions; Peptides; Phenotype; Physiological; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor beta Receptor; Process; Proliferating; Proteolysis; Proteolytic Processing; Pulmonary Fibrosis; Regulation; Role; Signal Pathway; Signal Transduction; Staging; System; Testing; Time; Tissues; Tyrosine; Up-Regulation; Wound Healing; attenuation; cancer type; cell growth regulation; cell motility; chronic wound; extracellular; healing; insight; novel; public health relevance; response; scaffold; success; tool; wound

 DESCRIPTION (provided by applicant): Platelet derived growth factor (PDGF) is the major mitogen for mesenchymal cells and is used clinically to promote diabetic ulcer healing. Clinical successes using PDGF to promote wound healing have been underwhelming and inconsistent. Fibronectin is a critical ECM protein that is secreted by fibroblasts in a protomeric form and assembled into a fibrillar matrix by a cell-mediated process termed fibronectin matrix assembly. Preliminary data presented in this proposal show that cell-assembled ECM fibronectin fibrils attenuate PDGF-induced intracellular calcium release. Intracellular calcium release was measured in Fluo-4- loaded fibronectin-null (FN-/-) mouse embryonic fibroblasts (MEFs), allowing for precise control over the amount of fibronectin to which cells were exposed and the initiation of fibronectin matrix assembly. Fibronectin- mediated attenuation of PDGF-induced calcium release represents a previously undescribed mechanism by which ECM fibronectin may downregulate PDGF signaling, and thus spatially and/or temporally control PDGF activity in healing wounds. Alternatively, several diseases including pulmonary fibrosis, atherosclerosis and some types of malignancies are characterized, in part, by an upregulation of PDGF activity. Thus, a fibronectin fragment that inhibits cell responses to PDGF may be beneficial in treating these conditions. The goal of this study is to determine the mechanism and consequences of ECM fibronectin-mediated regulation of cellular responses to PDGF in fibroblasts. In order to achieve the goals of this project, the following specific aims have been developed: (1) Localize the effect of ECM fibronectin on cell responsiveness to PDGF and develop small fibronectin peptides regulate cell responsiveness to PDGF, (2) determine the mechanism by which ECM fibronectin regulates fibroblast responses PDGF and (3) determine the effect of regulation of responses to PDGF by ECM fibronectin on cell phenotype. Important tools for these studies will include calcium analysis in our Fluo-4-loaded FN-/- MEF system, as well as our laboratory's extensive library of tools used to study fibronectin matrix assembly and expertise in developing novel fibronectin-derived fusion proteins. Experimental results will provide insight into how cell responses to PDGF are regulated during normal wound healing processes, how they may be dysregulated during pathological wound healing responses, and how specific fibronectin fragments may be used therapeutically to control endogenous PDGF activity.

 描述（由适用提供）：血小板衍生的生长因子（PDGF）是间充质细胞的主要有丝分裂原，可在临床上用于促进糖尿病性溃疡愈合。使用PDGF促进伤口愈合的临床成功一直是不一致和不一致的。纤连蛋白是一种关键的ECM蛋白，由原始形式的成纤维细胞分泌，并通过称为纤连蛋白基质组件的细胞介导的过程组装成原纤维基质。本提案中提供的初步数据表明，细胞组装的ECM纤连蛋白原纤维减弱了PDGF诱导的细胞内钙释放。在Fluo-4-载荷纤连蛋白 - 无（FN - / - ）小鼠胚胎纤连蛋白 - 无（MEFS）中测量细胞内钙释放，从而可以精确控制对细胞暴露于细胞的纤连蛋白量的精确控制以及纤连蛋白基质组组装的启动。纤连蛋白介导的PDGF诱导钙释放的衰减代表了一种先前不受调节的机制，通过该机制，ECM纤维蛋白可能会下调PDGF信号，因此在空间和/或暂时控制PDGF在愈合中的PDGF活性，而是多种疾病，包括肺纤维化，某些疾病，包括肺纤维化，分类的某些疾病，这些疾病是分类的，这些疾病的既定症状，这些疾病是类型的，一致性地构成了特征，这些疾病既有特征，一致性地构成了疾病，这些疾病既有特征，一致性地构成了特征。 活动。这是抑制细胞对PDGF反应的纤连蛋白片段可能对治疗这些疾病有益。这项研究的目的是确定ECM纤连蛋白介导的细胞对PDGF纤连蛋白中细胞反应的调节的机理和后果。为了实现该项目的目标，已经开发出以下具体目标：（1）将ECM纤连蛋白对PDGF细胞反应的作用定位，并开发小型纤连蛋白肽调节细胞对PDGF的反应性，（2）确定ECM纤维蛋白通过哪些ECM纤维素对PD的响应（3）效应（3）确定的机制（2）细胞表型上的纤连蛋白。这些研究的重要工具将包括我们加载的FN-4/ - MEF系统中的钙分析，以及我们实验室的广泛工具库，用于研究纤连蛋白基质组装和开发新型纤连蛋白衍生融合蛋白的专业知识。实验结果将为您提供有关在正常伤口愈合过程中如何调节细胞对PDGF的反应，在病理伤口愈合反应中如何失调的细胞反应，以及如何使用特定的纤连蛋白片段在治疗中使用特定的纤连蛋白片段来控制内源性PDGF活性。