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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在癌症转移期间也是如此。这种细胞外基质的重塑是通过结构的形成来调节的，称为侵袭性和靶向性分泌酶，称为基质金属蛋白酶(MMPs)。跨足类细胞外基质的延伸和降解依赖于协调的局部肌动蛋白聚合以及靶向分泌MMPs，最终导致细胞迁移。然而，对于介导靶向基质金属蛋白酶分泌的机制以及基质金属蛋白酶分泌和肌动蛋白是如何产生的，我们知之甚少在细胞迁移过程中，动态是协调的。我们最近确定Rab40b是TARGET的关键调节因子乳腺癌细胞中基质金属蛋白酶的分泌和侵袭性伸展。我们已经展示了Tks 5和Segf是Rab40b 结合蛋白。值得注意的是，Tks5和Sgef都是肌动蛋白，并且在介形虫中都是调节蛋白。此外，我们还证明了cullin-5蛋白也可以与Rab40b结合，并可能介导内向蛋白的泛素化/降解。基于所有这些数据，我们提出了以下具体建议假设：(1)Rab40b与Sgef结合协调肌动蛋白聚合和基质金属蛋白酶的分泌。 (2)cullin-5与Rab40b结合调节细胞外基质迁移的动力学。细胞迁移过程中的内向伸展/回缩。该项目的目标是直接测试这些假设。首先，我们将确定Rab40b和Sgef复合体在调节肌动蛋白动态过程中的作用在体外形成内足细胞和细胞迁移。第二，我们将阐明cullin-5结合到 Rab40b在终止不定足的形成和细胞迁移中的作用。第三，我们将测试Rab40b在调节中的作用肌动蛋白动力学与体内细胞迁移。为此，我们将在斑马鱼过程中使用神经脊细胞迁移发展为实验模型，使我们能够在活体中分析细胞迁移和细胞外基质重塑胚胎发育过程中的细胞。总之，这项研究的完成将确定新的管理和管理机制细胞迁移过程中极化膜转运、细胞骨架动力学和细胞外基质重塑的协调在发育和致癌方面。