Functional analysis of the WAVE3 gene

WAVE3基因的功能分析

基本信息

批准号：
7667938
负责人：
JOHN K COWELL
金额：
$ 27.93万
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/7667938
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

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：转移是癌症死亡的主要原因，已被证明是由特定遗传事件控制的。从主要部位到遥远位点的转移的复杂性质涉及许多表型，这使得该过程的遗传学非常复杂。细胞运动，迁移和入侵等各个过程对于此过程中的多种控制机制是至关重要的，对于了解转移和针对该过程设计疗法至关重要。我们已经表明，波基因的表达是体外和体内细胞侵袭和乳腺癌细胞转移的关键促进因。在人类原发性乳腺癌中，与低级肿瘤相比，晚期肿瘤中的波蛋白可显着调节。在使用MDA-MB-231细胞的转移小鼠模型中，波的siRNA敲低显着降低了转移的潜力。免疫细胞化学表明，波功能的丧失会影响与细胞运动相关的肌动蛋白结构的正常组织，并且通过与PIS激酶的相互作用来介导这种作用。波浪的敲低导致形成薄片的失败，导致观察到的受限细胞运动。在模型细胞系统中，波似乎通过通过p38途径调节MMP产生来控制侵袭表型。我们的初步数据证明了波浪如何控制细胞侵袭和迁移的机制，为这些表型的另一个关键调节剂提供了重要的线索，这些型号影响了体内转移。为了更充分地了解波的功能，确定该基因的调节剂控制及其所涉及的途径将很重要，因为这可能会为细胞侵袭的调节提供新的见解。在此应用中，我们提出了一系列蛋白质组学方法，以识别与波相互作用的蛋白质，以更好地了解其所涉及的途径。我们还提出了一系列使用siRNA和体细胞敲除的实验，以消除特定细胞中的波的功能，以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，以更深入地探究波功能的机理。由于波的表达促进了侵袭，因此它是针对转移的治疗的有吸引力的潜在靶标，毕竟这是大多数情况下癌症进展的致命阶段。因此，更好地理解有关波浪功能的细胞内调节途径，可以为治疗干预提供新的机会。