Molecular Mechanisms of Invadopodia Formation and Function

侵袭伪足形成和功能的分子机制

• 批准号: 7921869
• 负责人: Vira V Artym
• 金额: $ 7.85万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7921869
• 关键词: Actins; Adhesions; Automation; Biological Models; Cancer Biology; Cancer Etiology; Cell Adhesion; Cell Adhesion Molecules; Cell model; Cells; Cessation of life; Collaborations; Comprehensive Cancer Center; Core Assembly; Cytoskeleton; Data; Disseminated Malignant Neoplasm; Doctor of Philosophy; Electron Microscopy; Environment; Event; Extracellular Matrix; Extracellular Matrix Degradation; Filament; Goals; Image Analysis; Imagery; In Vitro; Integrins; Invaded; Joints; Knowledge; Laboratories; Lead; Letters; Life; MMP14 gene; Malignant - descriptor; Malignant Neoplasms; Mediating; Membrane; Mentors; Mentorship; Metalloproteases; Microscopy; Molecular; Monitor; National Institute of Dental and Craniofacial Research; Neoplasm Metastasis; Outcome; Peptide Hydrolases; Planet Mars; Postdoctoral Fellow; Protein Biochemistry; Protein Dynamics; Proteins; Proteolysis; Public Health; Publishing; Regulation; Regulatory Pathway; Reporting; Research; Resources; Role; Scientist; Signal Transduction; Signaling Protein; Site; Small Interfering RNA; Structure; System; Techniques; Technology; Testing; Therapeutic; Time; Total Internal Reflection Fluorescent; Training; Tumor Cell Biology; Tumor Cell Invasion; Universities; Vinculin; Work; base; cancer cell; cell motility; cell type; cellular imaging; human EMS1 protein; in vivo; innovation; insight; instrumentation; neoplastic cell; novel; paxillin; physical property; prevent; stem; trafficking; tumor

DESCRIPTION (provided by applicant): Tumor metastasis is the leading cause of cancer deaths. Metastatic cancer cells use invadopodia, specialized filament-like membrane protrusions, to degrade and invade through surrounding extracellular matrix (ECM). However, invadopodia-mediated invasion remains understudied, and little is known about the molecular mechanisms that govern invadopodia formation and function. My long-term goal is to elucidate mechanisms of invadopodia-mediated invasion. This application has the dual objective of: (1) providing training required for my transition from postdoctoral fellow to independent successful scientist in the field of cancer biology/metastasis, and (2) determining the cellular mechanisms of invadopodia induction upon tumor cell adhesion to ECM, which is the next logical step in pursuit of my research goal. The central hypothesis of this proposal is that invadopodia assembly is initiated by signaling events at unique cell adhesions of invading cancer cells that we call preinvadopodial adhesions. This hypothesis stems from strong preliminary data, and was formulated on the basis of the unique live-cell model system that I developed in our laboratory for simultaneous visualization and analysis of invadopodia formation and function in ECM degradation (Artym et al., Cancer Res. 66:3034-43, 2006). Using three-channel live-cell imaging, I recently discovered that first, structural cores of invadopodia rich in actin and cortactin are formed, and then, MT1-MMP, a metalloprotease required for invadopodia function, is accumulated at the invadopodia triggering ECM degradation. I have now observed invadopodia initiation and formation at specific adhesion sites, preinvadopodial adhesions. To test the central hypothesis and to accomplish the objective of this application, three specific aims are proposed: 1) Determine the mechanism of invadopodia regulation by p1 and (33 integrins; 2) Determine the function of vinculin in invadopodia formation; 3) Determine the role of ECM in regulation of invadopodia induction from preinvadopodial adhesions in physiologically relevant 3D in vitro system. Confocal and TIRF microscopy techniques in combination with siRNA technology, protein biochemistry, and electron microscopy will be used. The proposed work is innovative because it takes advantage of the real-time live-cell model developed by me in our laboratory that allows characterizing protein dynamics at invadopodia while simultaneously monitoring invadopodia assembly and function. The proposed research is significant because it will expand our knowledge of malignant transformation and provide potential targets for control of tumor cell invasion and metastasis. Relevance to Public Health: understanding the mechanisms governing the formation and function of invadopodia should provide insights into tumor cell biology that could lead to potential therapeutic approaches to cancer metastasis.

描述（由申请人提供）：肿瘤转移是癌症死亡的主要原因。转移性癌细胞使用侵袭伪足（invadopodia），即专门的细胞毒性样膜突起，通过周围的细胞外基质（ECM）降解和侵入。然而，入侵伪足介导的入侵仍然研究不足，并知之甚少的分子机制，支配入侵伪足的形成和功能。我的长期目标是阐明侵袭足介导的入侵机制。这个应用程序有双重目标：（1）为我从博士后研究员过渡到癌症生物学/转移领域的独立成功科学家提供所需的培训，以及（2）确定肿瘤细胞粘附到ECM时侵袭伪足诱导的细胞机制，这是追求我的研究目标的下一个逻辑步骤。这个提议的中心假设是，侵袭伪足组装是由侵袭癌细胞独特的细胞粘附（我们称之为前侵袭伪足粘附）上的信号事件启动的。这一假设源于强有力的初步数据，并制定了独特的活细胞模型系统的基础上，我在我们的实验室开发的同时可视化和入侵伪足的形成和ECM降解功能的分析 （Artym等人，Cancer Res. 66：3034-43，2006）。使用三通道活细胞成像，我最近发现，首先，形成了富含肌动蛋白和coronin的侵袭伪足的结构核心，然后，MT 1-MMP，侵袭伪足功能所需的金属蛋白酶，在侵袭伪足积累，触发ECM降解。我现在已经观察到在特定的粘连部位，即侵袭性足前粘连，侵袭性足的发生和形成。 为了检验中心假设并实现本申请的目的，提出了三个具体目标：1）确定由β 1和β 3整合素调节侵袭伪足的机制; 2）确定黏着斑蛋白在侵袭伪足形成中的功能; 3）确定ECM在生理学相关的3D体外系统中从侵袭伪足前粘连诱导侵袭伪足的调节中的作用。将使用共聚焦和TIRF显微镜技术结合siRNA技术、蛋白质生物化学和电子显微镜。拟议的工作是创新的，因为它利用了我在实验室开发的实时活细胞模型，该模型允许表征入侵伪足的蛋白质动力学，同时监测入侵伪足的组装和功能。该研究具有重要意义，因为它将扩大我们对恶性转化的认识，并为控制肿瘤细胞的侵袭和转移提供潜在的靶点。与公共卫生的相关性：了解侵袭伪足的形成和功能的机制将有助于深入了解肿瘤细胞生物学，从而为癌症转移提供潜在的治疗方法。

项目成果

Preparation of High-Density Fibrillar Collagen Matrices That Mimic Desmoplastic Tumor Stroma.

• DOI: 10.1002/0471143030.cb1019s70
• 发表时间: 2016-03-01
• 期刊: Current protocols in cell biology
• 作者: Artym VV

Dense fibrillar collagen is a potent inducer of invadopodia via a specific signaling network.

• DOI: 10.1083/jcb.201405099
• 发表时间: 2015-02-02
• 期刊: The Journal of cell biology
• 作者: Artym VV;Swatkoski S;Matsumoto K;Campbell CB;Petrie RJ;Dimitriadis EK;Li X;Mueller SC;Bugge TH;Gucek M;Yamada KM
• 通讯作者: Yamada KM

Tensin 2 modulates cell contractility in 3D collagen gels through the RhoGAP DLC1.

• DOI: 10.1002/jcb.22460
• 发表时间: 2010-03-01
• 期刊: JOURNAL OF CELLULAR BIOCHEMISTRY
• 影响因子: 4
• 作者: Clark, Katherine;Howe, Jonathan D.;Pullar, Christine E.;Green, J. Angelo;Artym, Vira V.;Yamada, Kenneth M.;Critchley, David R.
• 通讯作者: Critchley, David R.

Imaging cells in three-dimensional collagen matrix.

• DOI: 10.1002/0471143030.cb1018s48
• 发表时间: 2010-09
• 期刊: Current protocols in cell biology
• 作者: Artym, Vira V;Matsumoto, Kazue
• 通讯作者: Matsumoto, Kazue