Understanding How Invadosomes Breach Basement Membrane In Vivo

了解体内侵袭体如何突破基底膜

基本信息

批准号：
8221154
负责人：
David R Sherwood
金额：
$ 27.72万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-01 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8221154
关键词：
Actins Basement membrane Behavior Biological Process Caenorhabditis elegans Cancer cell line Cell Culture Techniques Cell membrane Cells Cues Development Developmental Process Disease Disseminated Malignant Neoplasm Endothelial Cells Epithelial Cells Extracellular Matrix F-Actin Fibroblasts Genes Genetic Goals Guanosine Triphosphate Phosphohydrolases Health Human Image Immunologic Surveillance In Vitro Infiltration Integrins Invaded Lead Leukocytes Life Malignant Neoplasms Mediating Membrane Mission Modeling Molecular Analysis Molecular Target Mutagenesis Neoplasm Metastasis Optics Pathway interactions Penetration Physiological Processes Process Regulation Research Role Signal Transduction Site Specific qualifier value Stereotyping Structure Subcellular structure Therapeutic Tissues Visual Work cancer cell cell type cellular imaging clinically relevant cofilin genetic analysis human disease in vivo in vivo Model macrophage netrin receptor novel tumor progression vasodilator-stimulated phosphoprotein

项目摘要

DESCRIPTION (provided by applicant): Basement membrane is a thin, dense, sheet-like extracellular matrix that encircles most tissues and provides structural support for epithelial and endothelial cells. Understanding how specialized invasive cells cross basement membrane is of profound importance to human health: Basement membrane transmigration underlies the dispersal of cells in many developmental processes, is required in leukocytes for immune surveillance and inappropriate manifestation of cell-invasive behavior underpins the development of metastatic cancers. F-actin-rich, cell membrane associated structures, termed invadosomes, were identified over three decades ago in normally invasive cell types and metastatic cancer cell lines. Invadosomes are thought to mediate the ability of cells to invade through basement membrane barriers. Owing to the difficulty of examining cell-invasive behavior in vivo, invadosomes have only been studied in vitro, where cell culture conditions do not recapitulate native environmental signals or matrix conditions. As a result, the relevance, regulation and potential functions of invadosomes in vivo remain one of the most critical gaps in our understanding of cell- invasive behavior. Anchor cell invasion in C. elegans is a simple, highly stereotyped in vivo model of cell invasion through basement membrane that uniquely combines single-cell visual and genetic analysis. Through the development of live-cell imaging approaches, our group has recently identified dynamic invadosome structures within the anchor cell that breach the basement membrane. We find that when one invadosome penetrates the BM, new invadosome formation ceases, and a single invasive protrusion matures from the infiltrated invadosome. Using optical highlighting of basement membrane components, we have found that this protrusion pushes the basement membrane aside to clear a path for invasion. The goal of this application is to uncover the mechanistic details of how these invadosomes are used to breach basement membrane. Our proposed research will combine live-cell imaging with genetic and molecular analysis to determine: (1) how key pathways that regulate anchor cell invasion coordinately regulate the formation, dynamics and function of invadosomes prior to and during BM penetration, (2) the role of netrin signaling in selecting a single invadosome for basement membrane invasion, (3) the function of the actin regulator Ena/VASP in promoting basement membrane gap expansion at the anchor cell-basement membrane contact points after invadosome penetration. Together, this work will identify new mechanisms underlying cell invasion and elucidate how they function together to breach basement membrane. This project is relevant to NIH's mission because it will lead to specific therapeutic strategies to control invasive behavior in diseases such as metastatic cancer. PUBLIC HEALTH RELEVANCE: Cell invasion through basement membrane barriers occurs during the dispersal of cells in many normal physiological processes. In human diseases such as metastastic cancer, cell invasion is one of the most clinically relevant yet least understood aspects of cancer progression. The proposed research will investigate the mechanisms that control the subcellular structures that allow invasive cells to penetrate through basement membrane barriers. This work will reveal new molecular targets and strategies to develop more effective and specific treatments to control cell invasive behavior in important human diseases such as cancer.

描述（由申请人提供）：地下膜是一种薄，密集的细胞外基质，它包围大多数组织，并为上皮和内皮细胞提供结构支持。了解专门的侵入性细胞穿越基底膜对人类健康至关重要：基底膜的迁移是细胞在许多发育过程中的扩散构成的，在白细胞中需要免疫监测和不适当的细胞侵入行为表现为转移性癌症的发育。三十年前，在正常浸润性细胞类型和转移性癌细胞系中，鉴定出富含F-肌动蛋白的细胞膜相关结构，称为Intivosomes。被认为可以介导细胞通过基底膜屏障侵袭细胞的能力。由于难以在体内检查细胞侵入性行为，因此仅在体外研究了侵袭体，在细胞培养条件下没有概括天然环境信号或基质条件。结果，体内侵袭体的相关性，调节和潜在功能仍然是我们对细胞侵袭行为的理解中最关键的差距之一。秀丽隐杆线虫中的锚细胞侵袭是通过基底膜的细胞侵袭模型简单，高度定型的，它独特地结合了单细胞视觉和遗传分析。通过开发活细胞成像方法，我们的小组最近确定了突破地下室膜的锚式细胞内的动态侵袭体结构。我们发现，当一个侵袭体穿透BM时，新的侵袭体形成停止，并且从浸润的侵袭体中的单个侵入性突出也成熟。使用基底膜组件的光学突出显示，我们发现这种突出将基底膜推开以清除入侵路径。该应用程序的目的是揭示如何使用这些侵袭体违反地下室膜的机械细节。 Our proposed research will combine live-cell imaging with genetic and molecular analysis to determine: (1) how key pathways that regulate anchor cell invasion coordinately regulate the formation, dynamics and function of invadosomes prior to and during BM penetration, (2) the role of netrin signaling in selecting a single invadosome for basement membrane invasion, (3) the function of the actin regulator Ena/VASP in promoting basement锚固细胞膜膜接触点的膜间隙膨胀在侵袭体渗透后的接触点。这项工作将共同确定细胞侵袭的新机制，并阐明它们如何共同发挥违反地下室膜。该项目与NIH的使命相关，因为它将导致特定的治疗策略来控制转移性癌症等疾病的侵入性行为。公共卫生相关性：在许多正常生理过程中细胞分散期间，通过基底膜屏障的细胞侵袭发生。在人类疾病（如转移癌症）中，细胞侵袭是临床上最相关的癌症进展方面最不明显的方面之一。拟议的研究将研究控制亚细胞结构的机制，从而使侵入性细胞穿过基底膜屏障。这项工作将揭示新的分子靶标和策略，以开发更有效和特定的治疗方法，以控制重要人类疾病（例如癌症）中的细胞侵入性行为。