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: 7186677
• 负责人: HILARY E BEGGS
• 金额: $ 34.42万
• 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 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7186677
• 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; Cyclophosphamide/Fluorouracil/Prednisone; Cytoskeleton; DNA Sequence Rearrangement; Defect; Deposition; Development; Disease; Disruption; Eels; Electroporation; Employee Strikes; Endopeptidases; Enzymes; Epithelial; Epithelial Cells; Event; Extracellular Matrix; Eye; Eye Development; Family member; Feedback; Focal Adhesion Kinase 1; Generations; Goals; Green Fluorescent Proteins; Heart; Histological Techniques; Image Analysis; Imagery; Immigration; Individual; Integrins; Investigation; Knock-out; Label; Laminin; Lens Fiber; Life; Mechanics; Microphthalmos; Modeling; Molecular; Morphology; Mus; Numbers; 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

DESCRIPTION: 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 functional 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 interaction 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 address 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 contribute 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.

描述：细胞基质粘附位点和细胞骨架之间的关键连接的丧失可能是许多疾病的核心。本项目的重点是阐明在发育中的小鼠透镜中调节这种功能性附着的分子机制。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和级联反应，包括1）细胞增殖，分化和迁移以及2）细胞外基质组织成基底膜。这种动态的双向相互作用的研究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眼透镜中的这些通路，因为它被体内最厚的基底膜之一包围，经历了确定的发育，可以完整地培养，并且是透明的，使其独特地可用于成像分析。我们将使用条件性敲除方法从发育中的小鼠透镜中删除FAK，其唯一的家族成员Pyk 2，以及整合素连接激酶（ILK）。 Pyk 2的缺失将解决功能冗余的问题，ILK的缺失将通过独立于FAK的替代途径破坏细胞-基质-细胞骨架附着来进一步测试我们的假设。我们将使用分子、细胞、生物化学和结构方法的组合来解决以下问题：目的1）FAK、Pyk 2和ILK信号传导是透镜发育所必需的吗？该通路的扰动是否会导致透镜病变？目的2）FAK相关信号的破坏是否导致透镜细胞迁移、增殖和/或分化的特定细胞缺陷？涉及哪些信号通路？目的3）FAK相关信号的破坏如何改变透镜囊基底膜的组织、合成和重塑？这些拟议的研究将提供细胞和细胞外基质之间所需的串扰，以及所涉及的信号转导途径的机制的见解。重要的是，这些结果也将提供独特的洞察眼睛发育的机制，并可能有助于更好地了解致盲性疾病，如小眼和先天性白内障，我们预测可能会导致错误的细胞基质相互作用在透镜细胞的发展。