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The mechanisms regulating actin dynamics and polarized membrane transport during invadopodia formation

侵袭伪足形成过程中调节肌动蛋白动力学和极化膜运输的机制

基本信息

批准号：
10092180
负责人：
Rytis Prekeris
金额：
$ 30.45万
依托单位：
UNIVERSITY OF COLORADO DENVER
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：
2018-02-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10092180
关键词：
3-Dimensional Actins Binding Binding Proteins Biological Assay Breast Cancer Cell Breast cancer metastasis Cancer Biology Cells Cellular biology Complex Cues Cullin 5 Protein Cytoskeleton Data Defect Development Embryonic Development Enzymes Experimental Models Extracellular Matrix Extracellular Matrix Degradation Foundations Goals Growth In Vitro Knowledge Laboratories Location Malignant Neoplasms Maps Matrix Metalloproteinases Mediating Microscopy Modeling Molecular Neoplasm Metastasis Neural Crest Cell Organ Organogenesis Pathway interactions Process Property Proteins Pseudopodia Publishing Research Role Shapes Structure Surface Testing Three-Dimensional Imaging Time Tissues Transmembrane Transport Ubiquitination Vesicle Work Zebrafish base cancer cell cancer therapy carcinogenesis cell motility design genetic regulatory protein in vivo in vivo Model migration multidisciplinary novel polymerization rab GTP-Binding Proteins recruit response two-dimensional zebrafish development

项目摘要

Project Summary Remodelling of the extracellular matrix (ECM) is a key process in cell migration during normal development as well as during cancer metastasis. This ECM remodelling is mediated via formation of structures, known as invadopodia and targeted secretion of enzymes, known as matrix metalloproteinases (MMPs). Invadopodia extension and degradation of ECM is dependent on coordinated localized actin polymerization as well as targeted secretion of MMPs at the tips of the invadopodia, which ultimately leads to cell migration. However, little is known about the mechanisms mediating targeted MMP secretion and how MMP secretion and actin dynamics are coordinated during cell migration. We recently identified Rab40b as a key regulator of targeted MMP secretion and invadopodia extension in breast cancer cells. We have shown Tks5 and SGEF are Rab40b binding proteins. Significantly, Tks5 and SGEF are both actin and in invadopodia regulating proteins. Additonally, we also demonstrated that Cullin-5 protein also binds to Rab40b and possibly mediate ubiquitination/degradation of invadopodial proteins. Based on all these data, we propose the following specific hypotheses: (1) Rab40b binding to SGEF coordinates actin polymerization and MMP secretion at the invadopodia during cell migration through the ECM; (2) Cullin-5 binding to Rab40b regulates dynamics of invadopodia extension/retraction during cell migration. The goal of this project is to directly test these hypotheses. First, we will define the roles of Rab40b and SGEF complex in regulating actin dynamics during invadopodia formation and cell migration in vitro. Second, we will elucidate the role of Cullin-5 binding to Rab40b in terminating invadopodia formation and cell migration. Third, we will test Rab40b role in regulating actin dynamics and cell migration in vivo. To that end, we will use neural crest cell migration during zebrafish development as experimental model that will allow us to analyse cell migration and ECM remodelling in live cells during embryogenesis. In summary, completion of this study will define new machinery governing and coordinating polarized membrane transport, cytoskeleton dynamics and ECM remodelling during cell migration in development and carcinogenesis.

项目摘要细胞外基质（ECM）的重塑是正常发育过程中细胞迁移的关键过程，以及在癌症转移期间。这种ECM重塑是通过形成称为侵袭伪足和靶向分泌的酶，称为基质金属蛋白酶（MMP）。侵袭伪足 ECM的延伸和降解依赖于协调的局部肌动蛋白聚合，在侵入伪足的尖端有针对性地分泌MMP，这最终导致细胞迁移。然而，在这方面，关于介导靶向MMP分泌的机制以及MMP分泌和肌动蛋白在细胞迁移期间协调动力学。我们最近确定Rab 40 b是靶向的乳腺癌细胞MMP分泌与侵袭伪足延伸。我们已经证明Tks 5和SGEF是Rab 40 b 结合蛋白值得注意的是，Tks 5和SGEF都是肌动蛋白和侵袭伪足调节蛋白。此外，我们还证明了Cullin-5蛋白也与Rab 40 b结合，并可能介导侵袭足蛋白的泛素化/降解。基于所有这些数据，我们提出以下具体建议假设：（1）Rab 40 b与SGEF的结合协调肌动蛋白聚合和MMP分泌，（2）Cullin-5与Rab 40 b的结合调节细胞迁移过程中的侵袭伪足动力学。在细胞迁移过程中侵入伪足延伸/收缩。这个项目的目标是直接测试这些假设首先，我们将确定Rab 40 b和SGEF复合物在调节肌动蛋白动力学过程中的作用。体外侵袭伪足形成和细胞迁移。第二，我们将阐明Cullin-5结合至 rab 40 b终止侵入伪足形成和细胞迁移。第三，我们将测试Rab 40 b在调节肌动蛋白动力学和体内细胞迁移。为此，我们将使用神经嵴细胞迁移在斑马鱼开发作为实验模型，这将使我们能够分析细胞迁移和ECM重塑在生活中胚胎发育过程中的细胞。总之，完成这项研究将确定新的机制，在细胞迁移过程中协调极化膜转运、细胞骨架动力学和ECM重塑在发育和致癌过程中的作用。