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

• 批准号: 7573094
• 负责人: RASHMI S. HEGDE
• 金额: $ 37.5万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 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/7573094
• 关键词: Adhesions; Adhesiveness; Adult; Age related macular degeneration; Area; Binding; Biochemical Genetics; Biological; Biological Assay; Cadherins; Cataract; Cell Adhesion; Cell Differentiation process; Cells; Complex; Crystalline Lens; Defect; Development; Diabetic Retinopathy; Disease; Down-Regulation; Embryo; Epithelial; Epithelial Cells; Epithelium; Eye; Eye Development; Fetal Development; Genetic; Genotype; Goals; Head; Hemorrhage; Infection; Integral Membrane Protein; Label; Lead; Left; Ligand Binding; Ligands; Mediating; Membrane Proteins; Mesenchyme; Methods; Modeling; Morphogenesis; Mus; Mutant Strains Mice; Neoplasm Metastasis; Phosphorylation; Process; Public Health; Recombinant Proteins; Regulation; Role; Sagittaria; Study Subject; Surface Ectoderm; System; Vascular Diseases; Vascular Endothelial Cell; Work; base; beta catenin; cadherin 5; epithelial to mesenchymal transition; experience; fiber cell; interest; lens; neoplastic cell; novel therapeutics; stoichiometry; tool; tumor; tumor progression; Adhesions; Adhesiveness; Adult; Age related macular degeneration; Area; Binding; Biochemical Genetics; Biological; Biological Assay; Cadherins; Cataract; Cell Adhesion; Cell Differentiation process; Cells; Complex; Crystalline Lens; Defect; Development; Diabetic Retinopathy; Disease; Down-Regulation; Embryo; Epithelial; Epithelial Cells; Epithelium; Eye; Eye Development; Fetal Development; Genetic; Genotype; Goals; Head; Hemorrhage; Infection; Integral Membrane Protein; Label; Lead; Left; Ligand Binding; Ligands; Mediating; Membrane Proteins; Mesenchyme; Methods; Modeling; Morphogenesis; Mus; Mutant Strains Mice; Neoplasm Metastasis; Phosphorylation; Process; Public Health; Recombinant Proteins; Regulation; Role; Sagittaria; Study Subject; Surface Ectoderm; System; Vascular Diseases; Vascular Endothelial Cell; Work; base; beta catenin; cadherin 5; epithelial to mesenchymal transition; experience; fiber cell; interest; lens; neoplastic cell; novel therapeutics; stoichiometry; tool; tumor; tumor progression

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展机制。在此应用中，我们更受限制的目标是确定不寻常的跨膜蛋白Crim1如何介导发育中的晶状体中的上皮粘附和形态发生。我们的初步分析表明，CRIM1通过与β-蛋白和钙粘着蛋白的相互作用来调节细胞粘附。 Crim1可以与B-catenin形成复合物，而B-catenin又与钙粘蛋白相互作用。我们的中心假设是，通过与Catenin和cadherin的复杂形成，Crim1调节上皮细胞粘附和形态发生。为了研究Crim1在镜头中的功能，我们提出了两个具体的目的：目标1。确定Crim1，β-catenin和Cadherin之间的相互作用机理。通过基于细胞或重组蛋白的相互作用测定，我们将确定CRIM1是否与任何已知的配体结合机理结合β-catenin。这将包括评估磷酸化的作用，因为众所周知，这会影响其他β-catenin配体的结合。我们还将确定Crim1-beta-catenin/cadherin复合物的化学计量法，因为这可能具有重要的生物学意义。目标2。确定Crim1-beta-catenin的相互作用是否调节镜片中的钙粘蛋白依赖性粘附。使用Crim1的形式，我们将无法与β-catenin相互作用，我们将使用有条件的突变小鼠来确定Crim1-beta-catenin相互作用是否对发育型晶状体中的钙粘着蛋白依赖性粘附很重要。对发育形态发生的研究本身很有趣，但对我们对疾病的理解产生了重要的影响。例如，钙粘蛋白粘附性的调节对于眼科特异性疾病（例如继发性白内障形成）至关重要，其中有间质转变和血管疾病的糖尿病性视网膜病变和与年龄相关的黄斑病变性，需要在血管内无内皮细胞中Ve-粘肽的调节。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，侵入性并最终形成转移酶。