A Novel RhoG Protein Interaction Network in Invadopodia

Invadopodia 中新型 RhoG 蛋白相互作用网络

• 批准号: 9318481
• 负责人: Rafael Garcia-Mata
• 金额: $ 7.56万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-19 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9318481
• 关键词: Actins; Behavior; Binding; Biological Assay; Blood Circulation; Breast Cancer Cell; Breast Cancer Patient; Cells; Cytoskeleton; Disseminated Malignant Neoplasm; Distant; Extracellular Matrix; Fostering; Future; Goals; Health; Invaded; Knowledge; Label; Light; Mass Spectrum Analysis; Mediating; Membrane; Mission; Neoplasm Metastasis; Organ; Pathway interactions; Play; Primary Neoplasm; Protein Family; Proteins; Proteomics; Public Health; Regulation; Research; Role; Signal Pathway; Signal Transduction; Subgroup; Testing; Time; Tissues; United States National Institutes of Health; Work; base; cancer cell; cell motility; cell type; improved; in vivo; migration; mortality; novel; protein function; protein protein interaction; rho GTP-Binding Proteins; tumor progression

PROJECT SUMMARY In breast cancer patients, the migration of cancer cells away from the primary tumor and their subsequent metastasis to distant organs is the leading cause of mortality. Metastatic cells escape the primary tumor and enter the bloodstream by developing actin-rich membrane protrusions called invadopodia that degrade the extracellular matrix (ECM) to allow invasion of surrounding tissues. The assembly of invadopodia is regulated by Rho GTPases, a family of proteins that regulates the actin cytoskeleton. Deregulation in Rho GTPase signaling has been associated with all stages of cancer progression, including proliferation, invasion and metastasis. However, little is known about how they are activated, the time course of their activation, or the identity of their upstream regulators and downstream effectors. Our long term goal is to characterize the mechanisms of regulation of Rho GTPases that contribute to cancer cell metastasis, in particular to cancer cell migration and invasion. Our preliminary results show that RhoG plays a key role in the regulation of invadopodia formation in breast cancer cells. Based on our preliminary results, our central hypothesis is that RhoG functions as a negative regulator of invadopodia formation. To characterize the RhoG signaling components involved in invadopodia formation, we have performed a proteomic analysis of the RhoG interactome and identified several potential RhoG binding partners. This application focuses on a subgroup of functionally related proteins, which includes potential upstream and downstream components involved in RhoG-mediated signaling. The objective of this application is to characterize these novel protein- protein interactions and their role during invadopodia formation and cell invasion. We will test our hypothesis by pursuing two specific aims: Aim 1. To validate the RhoG interaction partners identified by mass spectrometry. Using a proximity- based labeling assay we have identified a network of potential RhoG-interaction proteins. The goal of this aim is to validate the interactions identified by mass spectrometry. Aim 2. Characterization of the role of the identified proteins during invadopodia formation. We have uncovered a protein interaction network that may play a role in the regulation of RhoG-mediated invadopodia formation. The goal of this aim is to characterize the role of the different RhoG interaction partners on the regulation of RhoG activity, invadopodia formation, and cell invasion. It is expected that the knowledge generated from these studies will advance our understanding of how Rho GTPase pathways regulate critical steps during cancer progression. Future work will build up on this application and extend these studies to dissect the function of this protein network during cell invasion and metastasis in vivo.

项目摘要 在乳腺癌患者中，癌细胞远离原发肿瘤的迁移及其 随后转移到远处器官是死亡的主要原因。转移细胞逃脱了 原发性肿瘤，并通过发展富含肌动蛋白的膜突起进入血液， 侵袭伪足降解细胞外基质（ECM）以允许侵袭周围组织。的 侵入伪足的组装受Rho GTP酶的调节，Rho GTP酶是调节肌动蛋白的蛋白质家族， 细胞骨架Rho GT3信号的失调与癌症的所有阶段有关 进展，包括增殖、侵袭和转移。然而，人们对它们是如何工作的知之甚少。 被激活，其激活的时间过程，或其上游调控因子的身份， 下游效应器。我们的长期目标是表征Rho的调节机制， 促进癌细胞转移，特别是癌细胞迁移和侵袭的GTP酶。 我们的初步结果表明，RhoG在调节侵袭伪足的形成中起着关键作用。 乳腺癌细胞根据我们的初步结果，我们的中心假设是RhoG的功能是 侵入伪足形成的负调节因子。表征RhoG信号传导组分 我们对RhoG相互作用组进行了蛋白质组学分析， 并鉴定了几种潜在的RhoG结合伴侣。该应用程序集中在一个子组 功能相关的蛋白质，包括潜在的上游和下游成分， RhoG介导的信号传导。本申请的目的是表征这些新的蛋白质， 蛋白质相互作用及其在侵入伪足形成和细胞侵入过程中的作用。我们将测试我们的 通过追求两个具体目标的假设： 目标1.验证通过质谱鉴定的RhoG相互作用伴侣。利用一种接近- 基于标记分析，我们已经鉴定了潜在的RhoG相互作用蛋白的网络。的目标 目的是验证质谱法鉴定的相互作用。 目标2.鉴定的蛋白质在侵入伪足形成期间的作用的表征。我们有 揭示了一个蛋白质相互作用网络，可能在RhoG介导的调节中发挥作用。 内陷伪足形成该目的的目标是表征不同RhoG相互作用的作用。 在调节RhoG活性、侵袭伪足形成和细胞侵袭方面的作用。 预计这些研究产生的知识将促进我们的理解 Rho GT3通路如何调节癌症进展过程中的关键步骤。今后的工作将建立 在这个应用上，并扩展这些研究，以剖析这种蛋白质网络在细胞中的功能， 体内侵袭和转移。