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: 7500310
• 负责人: RYAN G MCALLISTER
• 金额: $ 14.79万
• 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/7500310
• 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（由申请人提供）：侵袭伪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。拟议研究的目标是确定侵袭伪足通过 3D 胶原蛋白网络参与乳腺癌细胞迁移，并确定侵袭伪足功能的哪些方面对于侵袭至关重要。在 3D 培养模型中，考生将 1) 测量类似侵入伪足的膜延伸，2) 确定膜延伸和侵入伪足复合体形成之间的关系，3) 开发细胞基质粘附、丝状伪足、侵入伪足以及施加在 ECM 和基质降解上的局部力之间相互联系的机制模型，4) 确定侵入伪足在反应中的作用 趋化肿瘤细胞对受控趋化剂梯度的影响。因此，候选人将建立一套独特的工具来定量模拟 3D 侵袭细胞行为，并且该研究应该对侵袭伪足在侵袭中的多方面作用产生新的见解。 拟议的工作为候选人提供了实现其职业转型目标的理想机会，方法是：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、转染以及使用体外细胞行为分析（重点关注细胞运动和侵袭），以及 3）与成功的癌细胞生物学家（包括导师 Susette Mueller 博士）进行互动。 隆巴迪综合癌症中心。 相关性：一些细胞表面特征被认为是迁移和侵袭所必需的：丝状伪足和片状伪足的形成、基质细胞粘附和侵袭伪足。然而，这些局部位点的粘附和基质降解对细胞侵袭的贡献尚未在 3D 细胞培养环境中得到充分研究。典型癌细胞行为的一些关键组成部分可能只能在 3D 环境中重现。这里提出的实验是离体和体内组织成像的重要先决条件，可以探索微环境的其他复杂效应的贡献。了解侵袭伪足对于改变和穿过细胞外基质的重要性可能会确定新的干预目标以防止迁移和转移，这最终将提高生存率。 3D 聚合物基质中的定量成像、特征跟踪和位移测量具有挑战性，需要研究人员（例如具有光学系统设计、图像分析以及定量实验设计和解释经验的候选人）。