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

基本信息

批准号：
7387367
负责人：
HILARY E BEGGS
金额：
$ 33.84万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
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/7387367
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

项目摘要

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位于许多疾病。该项目的重点是阐明调节这一点的分子机制发育中的小鼠镜头中的iM依恋。我们的中心假设是局灶性粘附激酶 ^fak）和相关信号分子将细胞 - 矩阵的附着转换为细胞骨架重排和一系列响应，包括1）细胞增殖，分化和迁移以及2）将细胞外基质组织到基底膜中。研究这种动态的双向 nteraction需要检查天然的三维组织结构。我们的一般策略是在小鼠眼镜中研究这些途径，因为它被最厚的地下室之一包围体内的膜经历了定义的发育，可以完整培养，并且是透明的，使其独特地访问成像分析。我们将使用有条件的淘汰方法删除 FAK是其唯一的家庭成员Pyk2，以及发育中的小鼠镜头的整合素连接激酶（ILK）。 pyk2的删除将解决功能冗余问题，删除ILK将进一步测试我们的通过通过替代路线破坏细胞 - 玛质 - 细胞骨架附着的假设，而不是独立于 fak。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）镜头开发需要FAK，PYK2和ILK信号传导？做这种途径的扰动导致镜头病理学？ AIM 2）确实会破坏FAK相关的信号传导结果在透镜细胞迁移，增殖和/或分化的特定细胞缺陷中？什么信号通路参与吗？目标3）与FAK相关信号的破坏如何改变镜头胶囊的地下室膜是组织，合成和重新建模的吗？这些提出的研究将提供机械深入了解细胞和细胞外基质之间所需的串扰以及信号通路涉及。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有可能更好地理解诸如微观心脏病和我们预测的先天性白内障可能是由于晶状体细胞期间的细胞基质相互作用而导致的发展。