Cadherin Mechanotransduction

钙粘蛋白机械传导

基本信息

批准号：
9498490
负责人：
Deborah E Leckband
金额：
$ 33.42万
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/9498490
关键词：
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 Dimensions 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 Testing Tissues Transducers alpha catenin base biophysical analysis cellular engineering cohesion dimer feeding human model innovation 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.

这项赠款建立在我们的新颖发现的基础上，上皮细胞连接处的e-钙黏着蛋白会翻译机械信号，通过表皮生长因子受体（EGFR）激活激酶级联反应。电子 - 辅助蛋白是一个上皮细胞 - 细胞连接处的必需粘合剂蛋白，以及电子钙粘蛋白络合物也将力转化为调节细胞形状和上皮屏障完整性。这些新发现表明电子钙粘蛋白力 - 转导还激活调节细胞增殖，形态发生和疾病的信号。广泛的目标该程序的是确定E-Cadherin对EGFR机械激活的初始步骤，并建立这种机制的更广泛的生理意义。我们的初步数据也证明了这一点力激活信号通路调节通过 E-钙粘蛋白复合物中的α-catenin。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的核心变换机械中的必要组成部分。在这个程序，特定目标1检验了机械刺激的电子钙粘着蛋白激活EGFR的假设磷酸化，通过触发推定的E-钙粘蛋白/EGFR复合物的破坏。特定的目标2将使用由合作者Hristova（Johns Hopkins）开发的基于创新荧光的方法论研究质膜上电子钙粘着蛋白和EGFR之间的直接相互作用。拟议的研究是基于大量初步数据，该数据揭示了直接的蛋白质蛋白质关联。生物物理研究将使用E-钙粘蛋白和EGFR突变体的子集建立该关联的分子要求。特定目标3将在三维有机的三维中测试这些发现的物理含义与Weaver（UCSF）合作的人类乳腺上皮组织的模型。研究将决定电子钙粘蛋白/EGFR复合物是否确实是与整合素协调至对组织力学的调子形态发生和恶性肿瘤。乳房上皮细胞的3D培养物设计以表达E-钙粘蛋白突变体（AIM 2）将确定E-钙粘蛋白/EGFR复合物的影响对基质刚度的函数的增殖，形态发生和侵袭的破坏。整联蛋白很好已知可以与EGFR协调以调节乳腺组织发育和肿瘤进展。这些研究在该力敏感网络中，有可能将电子钙粘着蛋白作为基本组成部分。