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Molecular Analysis of Signal Transduction in Cell Migration

细胞迁移信号转导的分子分析

基本信息

批准号：
7408651
负责人：
JUN-LIN GUAN
金额：
$ 27.92万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-08-01 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7408651
关键词：
1-Phosphatidylinositol 3-Kinase Affect Binding Biological Models Biological Process Cell Adhesion Cell physiology Cells Complex Condition Cytoplasmic Tail Dynamin Embryonic Development Epithelial Fibronectins Focal Adhesion Kinase 1 Focal Adhesions Funding Goals Grant Injection of therapeutic agent Integrins Knockout Mice Malignant Neoplasms Mammary gland Mediating Microfluidic Microchips Modeling Molecular Molecular Analysis Mus N-terminal Neoplasm Metastasis Nude Mice PH Domain Phosphatidylinositols Phosphorylation Phosphorylation Site Phosphotransferases Process Property Recruitment Activity Regulation Role Signal Pathway Signal Transduction Signaling Molecule Tail Veins Work Wound Healing cell motility cell transformation directional cell in vivo kinase inhibitor malignant breast neoplasm migration novel novel strategies response scaffold

项目摘要

The long term goal of the proposed studies is to understand the signal transduction mechanisms in the regulation of cell migration. Previous funding period focused on the role of focal adhesion kinase (FAK) and its downstream target Grb7 and a putative novel FAK inhibitor FIP200 in the regulation of cell migration. We identified and characterized Grb7 interaction with phosphoinositides through its PH domain and a role for Grb7 in mediating EphB1 stimulation of cell migration, showed the critical importance of focal contacts localization of the FAK signaling complexes, and characterized FIP200 inhibition of FAK activity and cell migration. We also found an interaction between FAK and N-WASP and showed that FAK phosphorylation of N-WASP regulates N-WASP subcellular localization and promotion of cell migration. In addition, we found an interaction between integrin p1 and 14-3-3p and studied the role of 14-3-3p in the regulation of cell spreading and migration, and an intra-molecular interaction between the N-terminal FERM-like domain and the kinase domain of FAK that regulate FAK activation in an auto-inhibitory mechanism. In preliminary studies, we identified an interaction between FAK and endophilin A2 and showed that endophilin A2 association with FAK and phosphorylation by FAK/Src complex regulated its interaction with dynamin and affected invasion of Src transformed cells. We are also in the process of creating mammary epithelial-specific FAK conditional knockout mice to study the role of FAK and its interaction with endophilin A2 in breast cancer metastasis in vivo. Lastly, to facilitate studies of directional cell migration on ECMgradients which better mimic in vivo conditions, we fabricated microfluidics devices to generate defined and quantitative FN gradients and showed directional cell migration on the gradients with novel properties. In this proposal, we will 1) determine the mechanism and role of FAK interaction with endophilin A2 in the regulation of cell migration and invasion, 2) investigate the potential role of FAK and its interaction with endophilin A2 in cancer metastasis in vivo, and 3) analyze the responses and roles of FAK and other signaling molecules in directional cell migration on defined FN gradients. These studies will enhance our understanding of the molecular mechanisms of signal transduction in cell migration and invasion which are critical factors in biological processes such as embryonic development, wound healing and cancer.

拟议研究的长期目标是了解信号转导机制细胞迁移的调节。之前的资助期重点关注粘着斑激酶 (FAK) 的作用和其下游靶标 Grb7 和假定的新型 FAK 抑制剂 FIP200 在细胞迁移调节中的作用。我们鉴定并表征了 Grb7 通过其 PH 结构域与磷酸肌醇的相互作用以及 Grb7 在介导 EphB1 刺激细胞迁移中显示了焦点接触的至关重要性 FAK 信号复合物的定位，并表征了 FIP200 对 FAK 活性和细胞的抑制迁移。我们还发现了 FAK 和 N-WASP 之间的相互作用，并表明 FAK 磷酸化 N-WASP 调节 N-WASP 亚细胞定位和促进细胞迁移。此外，我们还发现整合素 p1 和 14-3-3p 之间的相互作用，并研究了 14-3-3p 在细胞调节中的作用扩散和迁移，以及 N 端 FERM 样结构域和 FAK 的激酶结构域，以自动抑制机制调节 FAK 激活。在初步研究中，我们确定了 FAK 和内皮素 A2 之间的相互作用，并表明内皮素 A2 关联 FAK 和 FAK/Src 复合物的磷酸化调节其与动力的相互作用并受影响 Src 转化细胞的侵袭。我们还在创建乳腺上皮特异性 FAK 条件敲除小鼠研究 FAK 的作用及其与内皮素 A2 的相互作用在乳腺癌中的作用体内转移。最后，为了促进 ECM 梯度上定向细胞迁移的研究，这更好模拟体内条件，我们制造了微流体装置来生成定义和定量的 FN 梯度并显示出具有新颖特性的梯度上的定向细胞迁移。在这个提案中，我们 1）确定FAK与内亲素A2相互作用在细胞调节中的机制和作用迁移和侵袭，2) 研究 FAK 的潜在作用及其与内皮素 A2 的相互作用癌症体内转移，3) 分析 FAK 和其他信号分子在体内的反应和作用在定义的 FN 梯度上定向细胞迁移。这些研究将加深我们对细胞迁移和侵袭中信号转导的分子机制是细胞迁移和侵袭的关键因素生物过程，例如胚胎发育、伤口愈合和癌症。