Mechanisms of Cell-Matrix Interaction and Signaling in Lens Development

晶状体发育中细胞-基质相互作用和信号传导的机制

• 批准号: 7796620
• 负责人: HILARY E BEGGS
• 金额: $ 34.22万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7796620
• 关键词: Actin-Binding Protein; Actins; Address; Architecture; Basal lamina; Basement membrane; Binding; Biochemical; Bromodeoxyuridine; Bypass; Cataract; Cell Adhesion Molecules; Cell Proliferation; Cell physiology; Cell-Matrix Junction; Cells; Cellular Morphology; Collagen; Control Animal; Crystalline Lens; Cytoskeleton; DNA Sequence Rearrangement; Defect; Deposition; Development; Disease; Eels; Electroporation; Employee Strikes; Enzymes; Epithelial; Epithelial Cells; Event; Extracellular Matrix; Eye; Eye Development; Family member; Feedback; Focal Adhesion Kinase 1; Generations; Goals; Heart; Histological Techniques; Image Analysis; Imagery; Immigration; Individual; Integrins; Investigation; Knock-out; Label; Laminin; Lens Fiber; Life; Mechanics; Microphthalmos; Modeling; Molecular; Morphology; Mus; Pathology; Pathway interactions; Peptide Hydrolases; Phosphotransferases; Play; Proline; Protein Tyrosine Kinase; Proteins; Reporter; Research Personnel; Retinal Dysplasia; Role; Route; Rupture; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Staining method; Stains; Structure; Surface; System; Technology; Testing; Thick; Tissues; Translating; Western Blotting; body system; cell motility; congenital cataract; extracellular; fiber cell; in vivo; insight; integrin-linked kinase; lens; lens capsule; migration; mutant; programs; response; scaffold; success

Loss of the pivotal connection between cell matrix adhesion sites and the cytoskeleton may lie at the heart of a number of diseases. This project is focused on elucidating the molecular mechanisms regulating this Imctional attachment in the developing mouse lens. Our central hypothesis is that focal adhesion kinase ^FAK) and related signaling molecules translate cell-matrix attachment into cytoskeletal rearrangements and a cascade of responses, including 1) cellular proliferation, differentiation and migration as well as 2) organization of the extracellular matrix into basement membranes. Study of this dynamic, bi-directional nteraction requires examination of a native, three-dimensional tissue structure. Our general strategy is to study these pathways in the mouse ocular lens, since it is surrounded by one of the thickest basement membranes in the body, undergoes a defined development, can be cultured intact, and is transparent, rendering it uniquely accessible to imaging analysis. We will use a conditional knockout approach to delete FAK, its only family member Pyk2, as well as integrin-linked kinase (ILK) from the developing mouse lens. Deletion of Pyk2 will addresses issues of functional redundancy, and deletion of ILK will further test our hypothesis by disrupting cell-matrix-cytoskeletal attachment through an alternative route, independent of FAK. We will use a combination of molecular, cellular, biochemical and structural approaches to address the following questions: Aim 1) Are FAK, Pyk2 and ILK signaling required for lens development? Does perturbation of this pathway result in lens pathology? Aim 2) Does disruption of FAK-related signaling result in specific cellular defects in lens cell migration, proliferation and/or differentiation? What signaling pathways are involved? Aim 3) How does disruption of FAK-related signaling alter how the lens capsule basement membrane is organized, synthesized and re-modeled? These proposed studies will provide mechanistic insight into the crosstalk required between cells and the extracellular matrix, and the signaling pathways involved. Importantly, these results will also provide unique insight into the mechanisms of eye development and potentially contriubte to a better understanding of blinding disorders such as microphthalmia and congenital cataracts that we predict may result from faulty cell matrix interactions during lens cell development.

细胞基质粘附位点和细胞骨架之间关键连接的丧失可能是 多种疾病。该项目的重点是阐明调节这一现象的分子机制 正在发育的小鼠镜头中的情感依恋。我们的中心假设是粘着斑激酶 ^FAK）和相关信号分子将细胞基质附着转化为细胞骨架重排， 一系列反应，包括 1) 细胞增殖、分化和迁移以及 2) 将细胞外基质组织成基底膜。研究这种动态的、双向的 相互作用需要检查天然的三维组织结构。我们的总体策略是 研究小鼠晶状体中的这些通路，因为它被最厚的基底之一包围 体内的膜，经历明确的发育，可以完整培养，并且是透明的， 使其易于成像分析。我们将使用条件淘汰的方法来删除 FAK、其唯一的家族成员 Pyk2，以及来自发育中的小鼠晶状体的整合素连接激酶 (ILK)。 删除 Pyk2 将解决功能冗余的问题，删除 ILK 将进一步测试我们的 通过另一种途径破坏细胞-基质-细胞骨架附着的假设，独立于 错误。我们将结合分子、细胞、生化和结构方法来解决 以下问题： 目标 1) 晶状体开发是否需要 FAK、Pyk2 和 ILK 信号传导？做 该通路的扰动会导致晶状体病理学吗？目标 2) 是否会破坏 FAK 相关信号传导 晶状体细胞迁移、增殖和/或分化的特定细胞缺陷？有哪些信号通路 参与？目标 3) FAK 相关信号传导的破坏如何改变晶状体囊基底的方式 膜被组织、合成和重塑吗？这些拟议的研究将提供机制 深入了解细胞和细胞外基质之间所需的串扰以及信号传导途径 涉及。重要的是，这些结果还将为眼睛发育机制提供独特的见解 并可能有助于更好地了解致盲性疾病，例如小眼症和 我们预测先天性白内障可能是由于晶状体细胞期间细胞基质相互作用错误造成的 发展。