CRIM1-b-Catenin-Cadherin Interactions in Eye Development and Disease

CRIM1-b-连环蛋白-钙粘蛋白在眼睛发育和疾病中的相互作用

基本信息

批准号：
7843631
负责人：
RASHMI S. HEGDE
金额：
$ 37.34万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-01 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7843631
关键词：
Actins Adhesions Adhesiveness Adult Age related macular degeneration Binding Biochemical Genetics Biological Biological Assay Biological Models Budgets Cadherins Cataract Cell Adhesion Cells Complex Crystalline Lens Cytoskeleton Data Defect Development Diabetic Retinopathy Disease Down-Regulation Epithelial Epithelial Cells Epithelium Equilibrium Eye Eye Development Fetal Development Genetic Genetic Models Goals Infection Integral Membrane Protein Lead Learning Left Ligand Binding Ligands Mediating Methods Modeling Morphogenesis Mus Mutant Strains Mice Neoplasm Metastasis Organism Phosphorylation Process Public Health Recombinant Proteins Regulation Relative (related person)Role Staging Study Subject System Time Vascular Diseases Vascular Endothelial Cell Work base beta catenin cadherin 5 design epithelial to mesenchymal transition experience in vivo innovation interest lens lens morphogenesis neoplastic cell novel therapeutics protein protein interaction stoichiometry tool tumor tumor progression

项目摘要

Our long-term goal is an understanding of the mechanisms of lens development. In this application, our more restricted goal is to determine how the unusual transmembrane protein CRIM1 mediates epithelial adhesion and morphogenesis in the developing lens. Our preliminary analysis has shown that CRIM1 regulates cell adhesion through interactions with beta-catenin and cadherins. CRIM1 can form a complex with b-catenin which in turn interacts with cadherins. Our central hypothesis is that through complex formation with catenins and cadherins, CRIM1 regulates epithelial cell adhesion and morphogenesis. To investigate the function of CRIM1 in the lens, we propose two specific aims: Aim 1. To determine the mechanism of interaction between CRIM1, beta-catenin and cadherins. With interaction assays that are cell-based or recombinant protein-based we will determine whether CRIM1 binds beta-catenin with any of the known ligand binding mechanisms. This will include an assessment of the role of phosphorylation as this is known to influence binding of other beta-catenin ligands. We will also determine the stoichiometry of CRIM1-beta-catenin/cadherin complexes as this may have important biological implications. Aim 2. To determine whether the CRIM1-beta-catenin interaction regulates cadherin-dependent adhesion in the lens. Using forms of CRIM1 that either cannot or obligatorily interact with beta-catenin we will determine, using rescue of CRIM1 conditionally mutant mice, whether the CRIM1-beta-catenin interaction is important for cadherin dependent adhesion in the developing lens. The study of developmental morphogenesis is interesting in its own right but also has important consequences for our understanding of disease. For example, regulation of cadherin adhesiveness is critical for eye-specific diseases such as secondary cataract formation where there is an epithelial to mesenchymal transition and in the vascular diseases diabetic retinopathy and age-related macular degeneration that require the modulation of VE-cadherin in vascular endothelial cells. The down-regulation of cadherin expression and activity is also know to be a critical step in epithelial tumor progression as it allows tumor cells to leave the epithelium, become invasive and ultimately form metastases.

我们的长期目标是了解晶状体发育的机制。在此应用中，我们更严格的目标是确定不寻常的跨膜蛋白 CRIM1 如何介导发育中晶状体的上皮粘附和形态发生。我们的初步分析表明 CRIM1 通过与 β-连环蛋白和钙粘蛋白的相互作用来调节细胞粘附。 CRIM1 可以与 β-连环蛋白形成复合物，而 β-连环蛋白又与钙粘蛋白相互作用。我们的中心假设是，通过与连环蛋白和钙粘蛋白形成复合物，CRIM1 调节上皮细胞粘附和形态发生。为了研究 CRIM1 在晶状体中的功能，我们提出了两个具体目标：目标 1. 确定 CRIM1、β-连环蛋白和钙粘蛋白之间相互作用的机制。通过基于细胞或重组蛋白的相互作用测定，我们将确定 CRIM1 是否通过任何已知的配体结合机制结合 β-连环蛋白。这将包括对磷酸化作用的评估，因为已知磷酸化会影响其他 β-连环蛋白配体的结合。我们还将确定 CRIM1-β-连环蛋白/钙粘蛋白复合物的化学计量，因为这可能具有重要的生物学意义。目标 2. 确定 CRIM1-β-连环蛋白相互作用是否调节晶状体中钙粘蛋白依赖性粘附。使用不能或必须与β-连环蛋白相互作用的CRIM1形式，我们将通过拯救CRIM1条件突变小鼠来确定CRIM1-β-连环蛋白相互作用对于发育中的晶状体中的钙粘蛋白依赖性粘附是否重要。发育形态发生的研究本身就很有趣，而且对我们对疾病的理解也有重要影响。例如，钙粘蛋白粘附性的调节对于眼睛特异性疾病至关重要，例如继发性白内障形成，其中存在上皮到间质的转变，以及血管疾病糖尿病性视网膜病和年龄相关性黄斑变性，这些疾病需要调节血管中的VE-钙粘蛋白内皮细胞。钙粘蛋白表达和活性的下调也被认为是上皮肿瘤进展的关键步骤，因为它允许肿瘤细胞离开上皮，变得具有侵袭性并最终形成转移。