Biomechanics of Invading Breast Cancer Cells in 3D Matrices: Invadopodia Function

3D 矩阵中侵袭乳腺癌细胞的生物力学：侵袭伪足功能

• 批准号: 7384723
• 负责人: RYAN G MCALLISTER
• 金额: $ 14.69万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-28 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7384723
• 关键词: Actin-Binding Protein; Actins; Address; Adherence; Adhesions; Behavior; Binding; Biological Assay; Biomechanics; Breast Cancer Cell; Caliber; Cell Adhesion; Cell Surface Extensions; Cell surface; Cell-Matrix Junction; Cells; Cellular biology; Characteristics; Chemotactic Factors; Chemotaxis; Collagen; Complex; Comprehensive Cancer Center; Cultured Cells; Cytoskeleton; Depth; Development; Education; Endopeptidases; Environment; Experimental Designs; Extracellular Matrix; Extracellular Matrix Degradation; Face; Fiber; Filopodia; Focal Adhesions; Goals; Growth Factor; Ideal 1; Image; Image Analysis; Imaging Techniques; In Vitro; Intervention; Invaded; Invasive; Knowledge; Label; Localized; Location; MMP14 gene; Malignant Neoplasms; Measurement; Measures; Mediating; Membrane; Mentors; Methods; Microscope; Microscopic; Microscopy; Microspheres; Modeling; Molecular; Morphogenesis; Neoplasm Metastasis; Numbers; Optics; Pathway interactions; Penetration; Peptide Hydrolases; Phosphotransferases; Plasmids; Polymers; Process; Property; Proteolysis; Publishing; Quantum Dots; Relative (related person); Research; Research Design; Research Personnel; Resistance; Resolution; Role; Site; Small Interfering RNA; Speed; System; Techniques; Testing; Thinking; Tissues; Training; Transfection; Tumor Cell Invasion; Variant; Wasps; Work; base; cancer cell; career; cell behavior; cell motility; design; experience; human EMS1 protein; human MMP14 protein; improved; in vitro Assay; in vivo; indexing; insight; laser tweezer; migration; mutant; neoplastic cell; neuronal cell body; novel; polymerization; prevent; programs; protein aggregation; research study; response; rho; three-dimensional modeling; tool; tumor; tumor progression

DESCRIPTION (provided by applicant): Invadopodia are thin cell membrane extensions detected in aggressive tumor cells and defined by their role in localized extracellular matrix degradation. The goal of the proposed research is to determine the involvement of invadopodia in breast cancer cell migration through 3D collagen networks and to determine which aspects of invadopodia function are critical for invasion. In 3D culture models, the candidate will 1) measure invadopodia-like membrane extensions, 2) determine the relationship between membrane extension and invadopodial complex formation, 3) develop a mechanistic model of the interconnections between cell-matrix adhesions, filopodia, invadopodia and to localized force exerted on the ECM and matrix degradation, and 4) determine the role of invadopodia in the response of chemotactic tumor cells to controlled chemoattractant gradients. As a result, the candidate will have built a unique set of tools to quantitatively model invasive cell behavior in 3D, and the research should produce new insights into the multi-faceted role of invadopodia in invasion. The proposed work provides the candidate with an ideal opportunity to achieve his career transition goal by 1) education in cancer cell biology, 2) training in in vitro cell and molecular techniques including cell culture, manipulation of plasmids and siRNA, transfection, and use of in vitro assays of cellular behavior focusing on cell motility and invasion, and 3) interaction with successful cancer cell biologists, including the mentor Dr. Susette Mueller, in the setting of the Lombardi Comprehensive Cancer Center. Relevance: Several cell surface features are thought to be required for migration and invasion: filopodia and lamellipodia formation, matrix-cell adhesions, and invadopodia. Yet the contribution of adhesion and matrix degradation at these localized sites to cellular invasion has not been studied adequately in a 3D cell culture environment. Some critical components of typical cancer cell behavior can likely only be reproduced in the context of a 3D environment. Experiments proposed here are important prerequisites to imaging tissue ex vivo and in vivo where the contribution of other complex effects of the microenvironment can be explored. An understanding of the significance of invadopodia for altering and moving through the extracellular matrix may identify new targets for intervention to prevent migration and metastasis, which would ultimately improve survival. Quantitative imaging, feature-tracking, and displacement-measurement in a 3D polymer matrix is challenging and requires researchers such as the candidate with experience in optical system design, image analysis, and quantitative experimental design and interpretation.

描述(申请人提供)：侵袭足是在侵袭性肿瘤细胞中检测到的薄细胞膜延伸，并通过它们在局部细胞外基质降解中的作用来定义。这项研究的目的是确定内陷在乳腺癌细胞通过3D胶原蛋白网络的迁移中的作用，并确定内陷功能的哪些方面对侵袭至关重要。在3D培养模型中，候选者将1)测量内翻足细胞样膜延伸，2)确定膜延伸和内翻足复合体形成之间的关系，3)建立细胞-基质粘连、丝状足、内翻足与细胞外基质和基质降解的局部力之间的相互联系的机制模型，以及4)确定内翻足在趋化肿瘤细胞对可控制的化学吸引剂梯度的反应中的作用。因此，候选人将建立一套独特的工具，在3D中对侵袭细胞的行为进行定量建模，这项研究应该会对入侵足在侵袭中的多方面作用产生新的见解。 这项拟议的工作为候选人提供了一个理想的机会，通过1)癌细胞生物学教育，2)体外细胞和分子技术培训，包括细胞培养、质粒和siRNA操作、转基因和专注于细胞运动和侵袭的细胞行为体外分析的使用，以及3)在隆巴迪综合癌症中心与成功的癌细胞生物学家互动，包括与导师苏塞特·穆勒博士的互动。 相关性：几种细胞表面特征被认为是迁移和侵袭所必需的：丝足和片脂形成、基质-细胞粘连和内足。然而，在3D细胞培养环境中，这些局部部位的黏附和基质降解对细胞侵袭的贡献还没有得到充分的研究。典型癌细胞行为的一些关键成分很可能只能在3D环境中重现。这里提出的实验是对组织进行体外和体内成像的重要先决条件，在这里可以探索微环境的其他复杂效应的贡献。了解内毒素对细胞外基质改变和移动的重要性，可能会确定新的干预靶点，以防止迁移和转移，这最终将提高存活率。3D聚合物基质中的定量成像、特征跟踪和位移测量是具有挑战性的，需要具有光学系统设计、图像分析以及定量实验设计和解释经验的应聘者。