Role of beta-arrestins in chemokine receptor signaling

β-抑制蛋白在趋化因子受体信号传导中的作用

基本信息

批准号：
10391496
负责人：
Adriano Marchese
金额：
$ 32.05万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-02-11 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10391496
关键词：
Adaptor Signaling Protein Anatomy Attenuated Binding Biochemistry Biophysics CXCL12 gene Cell Culture Techniques Cell Survival Cell physiology Cell surface Cells Cellular biology Cessation of life Chemotaxis Complex Coupling Data Disease Disseminated Malignant Neoplasm Distant Focal Adhesion Kinase 1 Focal Adhesions G-Protein-Coupled Receptors GTP-Binding Protein alpha Subunits, Gs GTP-Binding Proteins Genetic Health Heterotrimeric GTP-Binding Proteins Homing In Vitro Integrins Knowledge Lead Ligands Link Liver Lung MAPK3 gene Malignant Neoplasms Mass Spectrum Analysis Mediating Metastatic to Modeling Molecular Molecular Biology Molecular Conformation Mutagenesis Pathway interactions Phosphorylation Play Process Prognosis Publishing RNA Interference Receptor Signaling Reporting Research Role Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Site Specific qualifier value Spectrum Analysis Stimulus Surface Techniques Testing Tissues Vesicle base beta-arrestin biophysical properties cancer cell cell motility chemokine chemokine receptor design innovation insight live cell imaging lymph nodes novel overexpression prevent protein B receptor receptor binding targeted treatment therapeutic target tumor progression

项目摘要

PROJECT SUMMARY The heterotrimeric G protein-coupled receptor (GPCR) C-X-C motif receptor 4 (CXCR4) and its cognate ligand CXCL12 play important roles in health and disease. A large body of evidence indicates that CXCR4 signaling is linked to cancer progression. CXCR4 expression and signaling in cancer correlates with poor prognosis2-5, mainly because cancer cells expressing CXCR4 colonize distant anatomical sites where CXCL12 is located, resulting in metastatic disease, the cause of most cancer related deaths. CXCR4 signaling regulates several aspects of cell physiology linked to cancer progression. This includes directed cell migration and cell survival, which occur via several discrete signaling pathways. Yet the mechanisms remain poorly understood. The focus of this proposal is on the signal transduction mechanisms that regulate CXCR4-mediated chemotaxis towards CXCL12. We recently reported that CXCR4-mediated chemotaxis occurs via a novel mechanism involving a complex formed between endocytic adaptor proteins b-arrestin1 (barr1) and STAM1 (barr1:STAM1). The barr1:STAM1 complex does not act on Akt or ERK-1/2 signaling pathways, but instead is necessary for activating focal adhesion kinase (FAK), which is also necessary for CXCL12 driven chemotaxis. FAK is typically linked to integrin signaling and focal adhesion dynamics, but these aspects of FAK function are not regulated by the barr1:STAM1 complex. Despite our contribution how barr1:STAM1 activates FAK downstream of CXCR4 to promote chemotaxis remains poorly understood. The overall objective of this proposal is to fill in knowledge gaps. Based on our published and preliminary studies we hypothesize that G protein-dependent barr1:STAM1 signaling spatially and temporally controls FAK activity required for CXCR4-dependent chemotaxis. To test this hypothesis we will pursue the following specific aims: Aim 1. To elucidate the role of CXCR4 site-specific phosphorylation on FAK activation; Aim 2. To identify the structural and biophysical properties of the barr1 interaction with STAM1; Aim 3. To elucidate the functional role of the barr1:STAM1 complex in chemotaxis. Because of the mechanistic focus of our proposal we will use cell culture models and other in vitro approaches spanning techniques in cell and molecular biology, genetics, biochemistry and biophysics plus advanced live cell imaging strategies and mass spectrometry approaches. At the conclusion of this project we will have learned novel signal transduction mechanisms by which barr1:STAM1 collaborate to activate FAK to promote chemotaxis. This is significant because it will reveal novel aspects of CXCR4 signaling that could be targeted therapeutically.

项目概要异三聚体 G 蛋白偶联受体 (GPCR) C-X-C 基序受体 4 (CXCR4) 及其同源配体 CXCL12 在健康和疾病中发挥着重要作用。大量证据表明 CXCR4 信号传导与癌症进展有关。癌症中的 CXCR4 表达和信号传导与不良预后相关2-5，主要是因为表达 CXCR4 的癌细胞定植于 CXCL12 所在的远处解剖部位，导致转移性疾病，这是大多数癌症相关死亡的原因。 CXCR4 信号调节多种与癌症进展相关的细胞生理学方面。这包括定向细胞迁移和细胞存活，这是通过几个离散的信号通路发生的。然而，人们对这些机制仍然知之甚少。焦点该提案的重点是调节 CXCR4 介导的趋化性的信号转导机制 CXCL12。我们最近报道，CXCR4 介导的趋化作用是通过一种新机制发生的，该机制涉及内吞接头蛋白 b-arrestin1 (barr1) 和 STAM1 (barr1:STAM1) 之间形成复合物。这 barr1：STAM1 复合物不作用于 Akt 或 ERK-1/2 信号通路，但对于激活粘着斑激酶 (FAK)，这对于 CXCL12 驱动的趋化性也是必需的。 FAK 是通常与整合素信号传导和粘着斑动力学相关，但 FAK 功能的这些方面并不受 barr1:STAM1 复合体调节。尽管我们做出了贡献，但 barr1:STAM1 是如何激活 FAK 下游的 CXCR4 促进趋化性的作用仍知之甚少。该提案的总体目标是填补知识差距。根据我们已发表的初步研究，我们假设 G 蛋白依赖性 barr1：STAM1 信号在空间和时间上控制 CXCR4 依赖性所需的 FAK 活性趋化性。为了检验这一假设，我们将追求以下具体目标：目标 1. 阐明 FAK 激活时 CXCR4 位点特异性磷酸化；目标 2. 确定结构和生物物理 barr1 与 STAM1 相互作用的特性；目标 3. 阐明 barr1:STAM1 的功能作用趋化性复杂。由于我们提案的机械重点，我们将使用细胞培养模型和其他体外方法涵盖细胞和分子生物学、遗传学、生物化学和生物物理学加上先进的活细胞成像策略和质谱方法。结束时在这个项目中，我们将了解 barr1:STAM1 合作的新颖信号转导机制激活FAK以促进趋化作用。这很重要，因为它将揭示 CXCR4 的新颖方面可以作为治疗目标的信号。