Cadherin Mechanotransduction

钙粘蛋白机械传导

基本信息

批准号：
9976560
负责人：
Deborah E Leckband
金额：
$ 32.94万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-05 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9976560
关键词：
3-Dimensional Acinus organ component Actins Adhesions Back Binding Biophysics Bioreactors Breast Epithelial Cells Cadherin Domain Cadherins Cell Proliferation Cell Shape Cell membrane Cells Cellular Stress Collaborations Complex Couples Coupling Data Dependence Development Disease E-Cadherin EGF gene Epidermal Growth Factor Receptor Epithelial Epithelial Cell Junction Epithelial Cells Epithelium Feedback Fluorescence Fluorescence Resonance Energy Transfer Goals Grant Integrins Intercellular Junctions MAP Kinase Gene Malignant Neoplasms Mammary Gland Parenchyma Mammary gland Measurement Mechanical Stimulation Mechanics Mediating Methodology Mitogen-Activated Protein Kinases Molecular Morphogenesis Mutation Pathway interactions Phosphorylation Phosphotransferases Physiological Proteins Psychological reinforcement Receptor Signaling Role Signal Pathway Signal Transduction Structure Testing Tissues Transducers base biophysical analysis cellular engineering cohesion dimer human model innovation mammary epithelium mechanical force mechanotransduction migration monomer mutant novel programs receptor binding response rho three dimensional cell culture tissue culture tumor progression two-dimensional

项目摘要

This grant builds on our novel discovery that E-cadherin at epithelial cell-cell junctions transduces mechanical signals, by activating a kinase cascade via the epidermal growth factor receptor (EGFR). E-cadherin is an essential adhesion protein at epithelial cell-cell junctions, and E-cadherin complexes also transduce force, to regulate cell shape and epithelial barrier integrity. These new findings suggest that E-cadherin force- transduction also activates signals that regulate cell proliferation, morphogenesis, and disease. The broad goal of this program is to identify initial steps in the mechanical activation of EGFR by E-cadherin, and to establish the broader physiological implications of this mechanism. Our preliminary data also demonstrate that this force-activated signaling pathway regulates the cytoskeletal reinforcement of stressed cell-cell junctions by α−catenin in E-cadherin complexes. This unexpected finding supports the hypothesis that EGFR and E- cadherin are essential components in the core force-transduction machinery at epithelial cell junctions. In this program, Specific Aim 1 tests the hypothesis that mechanically stimulated E-cadherin activates EGFR phosphorylation, by triggering the disruption of putative E-cadherin/EGFR complexes. Specific Aim 2 will use innovative fluorescence-based methodology, developed by collaborator Hristova (Johns Hopkins) to investigate direct interactions between E-cadherin and EGFR at the plasma membrane. Proposed studies are based on substantial preliminary data, which reveal direct protein-protein association. Biophysical studies will establish the molecular requirements for this association, using a subset of E-cadherin and EGFR mutants. Specific Aim 3 will test the physiological implications of these findings in a three-dimensional, organotypic model of human mammary epithelial tissue, in collaboration with Weaver (UCSF). Studies will determine whether E-cadherin/EGFR complexes are indeed central force-sensing units that coordinate with integrins to tune morphogenesis and malignancy, in response to tissue mechanics. 3D cultures of breast epithelial cells engineered to express E-cadherin mutants (Aim 2) will determine the impact of E-cadherin/EGFR complex disruption on proliferation, morphogenesis, and invasion, as a function of matrix rigidity. Integrins are well known to coordinate with EGFR to regulate breast tissue development and tumor progression. These studies would potentially establish E-cadherin as an essential component in this force-sensitive network.

这项资助建立在我们的新发现之上，即上皮细胞-细胞连接处的 E-钙粘蛋白转导机械作用信号，通过表皮生长因子受体 (EGFR) 激活激酶级联。上皮细胞-细胞连接处的必需粘附蛋白和 E-钙粘蛋白复合物也可以传导力，以调节细胞形状和上皮屏障完整性这些新发现表明 E-钙粘蛋白强制- 转导还激活调节细胞增殖、形态发生和疾病的信号。该计划的目的是确定 E-钙粘蛋白机械激活 EGFR 的初始步骤，并建立我们的初步数据也证明了这一机制的更广泛的生理意义。力激活信号通路通过以下方式调节应激细胞-细胞连接的细胞骨架强化 E-钙粘蛋白复合物中的 α-连环蛋白这一意外发现支持了 EGFR 和 E- 的假设。钙粘蛋白是上皮细胞连接处核心力传导机制的重要组成部分。程序，具体目标 1 测试机械刺激 E-钙粘蛋白激活 EGFR 的假设具体目标 2 将使用通过触发假定的 E-钙粘蛋白/EGFR 复合物的破坏来磷酸化。由合作者 Hristova（约翰霍普金斯大学）开发的基于荧光的创新方法研究 E-钙粘蛋白和 EGFR 在质膜上的直接相互作用。基于大量初步数据，这些数据揭示了蛋白质与蛋白质之间的直接关联。使用 E-钙粘蛋白和 EGFR 突变体的子集建立这种关联的分子要求。具体目标 3 将在三维、器官型模型中测试这些发现的生理学意义。人类乳腺上皮组织模型，将与韦弗研究（加州大学旧金山分校）合作确定。 E-钙粘蛋白/EGFR复合物是否确实是与整合素协调的中心力传感单元根据乳腺上皮细胞的 3D 培养物调整形态发生和恶性肿瘤。设计表达 E-钙粘蛋白突变体（目标 2）将确定 E-钙粘蛋白/EGFR 复合物的影响作为基质刚性的函数，对增殖、形态发生和侵袭的破坏是良好的。这些研究已知与 EGFR 协调调节乳腺组织发育和肿瘤进展。将建立潜在的E-钙粘蛋白作为这个力敏感网络的重要组成部分。