Fibronectin-dependent mechanisms governing the assembly of a definitive extracellular matrix

纤连蛋白依赖性机制控制最终细胞外基质的组装

• 批准号: 9923444
• 负责人: Jean E Schwarzbauer
• 金额: $ 34.55万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9923444
• 关键词: Address; Affect; Arthritis; Binding; Binding Proteins; Binding Sites; Biological Assay; Biological Models; Cell Differentiation process; Cell physiology; Cells; Chondrogenesis; Collagen; Collagen Fibril; Collagen Type I; Connective Tissue Diseases; Data; Defect; Development; Disease; Disease Outcome; Elements; Engineering; Enzymes; Extracellular Matrix; Extracellular Matrix Proteins; Fiber; Fibronectins; Fibrosis; Foundations; Glycosaminoglycans; Goals; Heparin; Heparitin Sulfate; Human; In Vitro; Knowledge; Lead; Link; Malignant Neoplasms; Mediating; Molecular; Molecular Conformation; Mutation; Normal tissue morphology; Pathogenicity; Pathologic Processes; Pathology; Patients; Process; Procollagen; Production; Property; Proteins; Pulmonary Fibrosis; Recombinant Proteins; Regulation; Research; Role; Route; Structure; System; Testing; Tissues; Work; base; chemical property; design; experimental study; fibrillogenesis; glomerulosclerosis; human disease; improved; insight; mechanical properties; mutant; novel; physical property; protein protein interaction; receptor; skeletal abnormality; skeletal dysplasia; tissue repair; treatment strategy; tumorigenesis

PROJECT SUMMARY The proposed work seeks to uncover novel mechanisms governing the assembly of the extracellular matrix (ECM) protein fibronectin (FN) and its role in directing assembly of other ECM proteins to form a definitive matrix. Because the definitive matrix is integral to key physical, chemical, and mechanical properties that regulate tissue structures and functions, understanding mechanisms that coordinate the assembly of FN with type I collagen and other ECM proteins is crucial. Molecular defects in matrix assembly are implicated in fibrosis in which disordered ECM fibers accumulate due to abnormal production of FN, collagen I, and other matrix proteins. Along with fibrosis, skeletal abnormalities, tumorigenesis, and other ECM-related diseases affect millions of people around the globe yet in most cases the ECM defects are poorly understood and, in many ways, still largely untreatable. The proposed work will test the hypothesis that the organization and insolubility of the pericellular FN matrix control and direct the assembly of a tissue-appropriate definitive matrix, and that perturbation of the FN matrix disrupts tissue and cell functions leading to disease. We have a general understanding of the main steps of FN matrix assembly, but specific mechanisms governing FN fibril organization, fibril stability, or how FN guides assembly of collagens have yet to be elucidated. The proposed aims will address these mechanisms, building on the foundation that we have established with our previous work. We will use our proven matrix assembly systems to analyze the formation of FN fibrils and their contributions to type I collagen assembly to determine the protein interactions that are critical for directing definitive matrix assembly. The goal of Aim 1 is to determine the mechanism that converts reversible FN-FN interactions into stable insoluble fibrils. We will test a new hypothesis that heparin/heparan sulfate acts as a molecular switch by binding to FN and inducing conformational changes that promote strong protein-protein interactions. Aim 2 will address the hypothesis that FN matrix acts as a template for collagen fibrillogenesis by providing a platform for the localization and activation of collagen processing enzymes. Aim 3 will link FN assembly with cell differentiation. Using a newly discovered human FN mutation potentially linked to a skeletal dysplasia, we will determine the matrix assembly defect caused by this mutation and will apply the micromass chondrogenesis model system to understand the effects of mutant FN matrix on cell differentiation. Our work will fill critical knowledge gaps in our understanding of the mechanisms governing key steps in the assembly of a definitive matrix, and will suggest routes by which pathological processes or mutations can cause abnormal matrix organization or accumulation. This work will generate new ideas for strategies to manipulate ECM assembly in order to treat or control fibrosis and other ECM-related diseases and may lead to ECM-based treatments to improve disease outcomes.

项目摘要 拟议的工作旨在揭示有关细胞外组装的新型机制 基质（ECM）蛋白质纤连蛋白（FN）及其在指导其他ECM蛋白的组装中的作用 确定的矩阵。因为确定的矩阵是关键物理，化学和机械性能不可或缺的 调节组织结构和功能，了解协调FN组装的机制 使用I型胶原蛋白和其他ECM蛋白至关重要。基质组件中的分子缺陷与 由于FN，胶原蛋白I和其他其他因素产生异常的ECM纤维累积的纤维化 基质蛋白。以及纤维化，骨骼异常，肿瘤发生和其他与ECM有关的疾病 影响全球数百万的人，但在大多数情况下，ECM缺陷知之甚少，并且在 许多方面，仍然很少无法治疗。拟议的工作将检验组织和组织的假设 细胞周围FN基质对照的不溶性，并指导适合组织的确定基质组装 FN基质的扰动破坏了组织和细胞功能导致疾病。我们有一个将军 了解FN矩阵组件的主要步骤，但是管理FN原纤维的特定机制 组织，原纤维稳定性或FN指南胶原蛋白的组装尚未阐明。提议 目的将解决这些机制，建立在我们与以前建立的基础之上 工作。我们将使用我们经过验证的矩阵组装系统来分析FN原纤维及其的形成 对I型胶原蛋白组装的贡献，以确定指导至关重要的蛋白质相互作用 确定的矩阵组件。目标1的目的是确定转换可逆FN-FN的机制 相互作用成稳定的不溶性原纤维。我们将检验一个新的假设，即肝素/硫酸乙酰肝素充当 通过与FN结合并诱导构象变化来促进强蛋白蛋白质的变化，分子转换 互动。 AIM 2将解决以下假设：FN矩阵充当胶原蛋白原纤维生成的模板 为胶原蛋白加工酶的定位和激活提供了一个平台。 AIM 3将链接FN 带有细胞分化的组装。使用新发现的人FN突变可能与骨骼有关 发育不良，我们将确定由此突变引起的基质组件缺陷，并将应用微瘤 软骨发生模型系统了解突变体FN基质对细胞分化的影响。我们的工作 将在我们理解有关​​集会中关键步骤的机制中填补关键知识差距 确定的矩阵，并建议通过病理过程或突变引起异常的路线 矩阵组织或积累。这项工作将为操纵ECM的策略产生新的想法 组装以治疗或控制纤维化和其他与ECM相关的疾病，并可能导致基于ECM的疾病 改善疾病预后的治疗方法。