Functional analysis of the WAVE3 gene

WAVE3基因的功能分析

基本信息

批准号：
8132654
负责人：
JOHN K COWELL
金额：
$ 11.76万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-06 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8132654
关键词：
Actins Affect Animals Antibodies Biological Assay Biological Process Blood Breast Cancer Cell Cell model Cell physiology Cells Cessation of life Co-Immunoprecipitations Complex Cytoskeleton Data Diagnostic Neoplasm Staging Distant Dominant-Negative Mutation Down-Regulation Event Failure Fiber Focal Adhesions Genes Genetic Genetic Processes Goals Human Immunohistochemistry In Vitro Individual Interstitial Collagenase Invaded Knock-out Lung Lung Neoplasms Lymph MAPK14 gene Malignant Neoplasms Mass Spectrum Analysis Matrix Metalloproteinases Mediating Nature Neoplasm Metastasis Nodule Null Lymphocytes Pathological Staging Pathway interactions Pattern Phase Phenotype Phosphorylation Phosphorylation Site Phosphotransferases Play Procedures Process Production Property Proteins Proteomics Regulation Regulatory Pathway Research Personnel Role SCID Mice Series Signal Pathway Site Small Interfering RNA Somatic Cell Staging Staining method Stains Structure System Tail Therapeutic Intervention Tissues Tumor Cell Invasion Tumor stage Up-Regulation Veins Yeasts cancer cell cell growth regulation cell motility cell type design human disease immunocytochemistry in vivo inhibitor/antagonist insight malignant breast neoplasm migration mitogen-activated protein kinase p38 mouse model neoplastic cell polymerization prevent programs protein protein interaction research study small hairpin RNA therapy design tumor tumor progression yeast two hybrid system

项目摘要

Metastasis is main reason for cancer death and has been shown to be controlled by specific genetic events. The complex nature of metastasis from the primary site to a distant site involves acquisition of many phenotypes which makes the genetics of the process very complex. Individual processes such as cell movement, migration and invasion are central to the process and understanding the multitude of controlling mechanisms in this process is vital to understand metastasis and designing therapies against the process. We have shown that expression of the WAVES gene is a critical facilitator of in vitro and in vivo cell invasion and metastasis in breast cancer cells. In human primary breast cancer, significant up regulation of the WAVES protein is seen in advanced stage tumors compared with low grade tumors. In a mouse model of metastasis using MDA-MB-231 cells, siRNA knockdown of WAVES significantly reduces metastasis potential. Immunocytochemistry shows that loss of WAVES function affects the normal organization of actin structures that are related to cell movement and that this effect is mediated through an interaction with PIS Kinase. Knockdown of WAVES results in a failure of lamellipodia formation which is responsible for the restricted cell movement observed. In model cell systems, WAVES appears to control the invasion phenotype through the regulation of MMP production through the p38 pathway. Our preliminary data demonstrating the mechanism of how WAVES controls cellular invasion and migration has provided important clues to another key regulator of these phenotypes which impact on metastasis in vivo. To understand the function of WAVES more fully it will be important therefore to determine the regulator controls of this gene and the pathways it is involved in, since this will likely provide new insights into the regulation of cellular invasion. In this application we propose a series of proteomics approaches to identify the proteins that interact with WAVES to obtain a better understanding of the pathways it is involved in. We also propose a series of experiments using siRNA and somatic cell knockout to eliminate WAVES function in specific cells to evaluate the importance of specific protein-protein interactions. Finally we have designed a series of experiments to probe more deeply into the mechanisms of WAVES function. Since expression of WAVES promotes invasion, it is an attractive potential target for therapy against metastasis, which after all is the lethal phase of cancer progression in most cases. A better understanding of the intracellular regulatory pathways governing the function of WAVES, therefore, may provide new opportunities for therapeutic intervention.

转移是癌症死亡的主要原因，已被证明受特定遗传事件的控制。从主要地点到远处的转移的复杂性质涉及许多使过程的遗传学非常复杂的表型。单个过程，例如单元运动，迁移和入侵是过程的核心，并了解控制的众多此过程中的机制对于了解转移和针对该过程的疗法设计至关重要。我们已经表明，波基因的表达是体外和体内侵袭的关键促进因子和乳腺癌细胞中的转移。在人类原发性乳腺癌中，对与低级肿瘤相比，在晚期肿瘤中可以看到波蛋白。在鼠标模型中使用MDA-MB-231细胞转移，波的siRNA敲低显着降低了转移潜在的。免疫细胞化学表明，波功能的损失会影响肌动蛋白的正常组织与细胞运动相关的结构，这种效应是通过与PIS的相互作用介导的激酶。波浪的敲低导致层状形成的失败，这是负责的观察到的受限制细胞运动。在模型细胞系统中，波似乎控制着侵袭通过通过p38途径调节MMP产生的表型。我们的初步数据证明波浪如何控制细胞侵袭和迁移的机制已提供这些表型的另一个关键调节剂的重要线索影响了体内转移。到更充分地了解波的功能，因此要确定调节器控制很重要该基因及其所参与的途径，因为这可能会提供有关调节的新见解细胞入侵。在此应用中，我们提出了一系列蛋白质组学方法来识别蛋白质与波相互作用，以更好地了解其所涉及的途径。我们也提出一系列使用siRNA和体细胞敲除以消除特定细胞中波的功能的实验评估特定蛋白质蛋白质相互作用的重要性。最后，我们设计了一系列实验更深入地探究波功能的机理。由于波的表达促进入侵，它是针对转移治疗的有吸引力的潜在目标，毕竟是在大多数情况下，癌症进展的致命期。更好地了解细胞内调节因此，管理波浪功能的途径可能会为治疗提供新的机会干涉。