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。该ECM重塑是通过形成结构的介导的，称为 Invadopodia和靶向分泌酶，称为基质金属蛋白酶（MMP）。 Invadopodia ECM的扩展和降解取决于协调的局部肌动蛋白聚合以及 MMP的靶向分泌在Invadopodia的尖端，最终导致细胞迁移。然而，关于介导的靶向MMP分泌的机制以及MMP分泌和肌动蛋白的介导的机制知之甚少在细胞迁移过程中，动力学是协调的。我们最近将Rab40b确定为目标的关键调节器乳腺癌细胞中的MMP分泌和侵袭性延伸。我们已经显示TKS5和SGEF为RAB40B 结合蛋白。值得注意的是，TKS5和SGEF既是肌动蛋白，又是调节蛋白质的Invadodia。添加剂，我们还证明了Cullin-5蛋白也与Rab40b结合，并可能介导侵袭性蛋白的泛素化/降解。基于所有这些数据，我们提出以下特定假设：（1）Rab40b与SGEF坐标结合肌动蛋白聚合和MMP分泌。细胞通过ECM迁移期间的Invadopodia；（2）Cullin-5与RAB40B结合调节动力学细胞迁移期间的Invadodia延伸/缩回。该项目的目的是直接测试这些假设。首先，我们将定义RAB40B和SGEF复合物在调节肌动蛋白动力学过程中的作用体外的侵袭性形成和细胞迁移。其次，我们将阐明Cullin-5结合与 Rab40b终止了Invadopodia形成和细胞迁移。第三，我们将测试RAB40B在调节中的作用肌动蛋白动力学和体内细胞迁移。为此，我们将在斑马鱼期间使用神经rest细胞迁移作为实验模型的开发将使我们能够分析现场的细胞迁移和ECM重塑胚胎发生过程中的细胞。总而言之，这项研究的完成将定义新机械管理和在细胞迁移期间，在细胞迁移过程中协调偏光膜传输，细胞骨架动力学和ECM重塑在发育和致癌作用中。