Array Microscope Assay for Cancer Cell Mechanics

癌细胞力学的阵列显微镜分析

• 批准号: 8154990
• 负责人: RICHARD SUPERFINE
• 金额: $ 32.24万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8154990
• 关键词: Address; Adhesions; Affect; Apoptotic; Artificial Heart; Ascites; Automobile Driving; Basic Science; Behavior; Biochemical Genetics; Biochemical Pathway; Biological Assay; Biological Markers; Biomechanics; Bone Morphogenetic Proteins; Breast; Cancer Model; Cancer cell line; Cancerous; Cell Line; Cell Mobility; Cell division; Cell membrane; Cell model; Cells; Cellular biology; Characteristics; Clinical; Cytoskeleton; Data Analyses; Development; Diagnosis; Diagnostic; Diffusion; Disease; Disseminated Malignant Neoplasm; Environment; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Extracellular Matrix; Genetic Determinism; Goals; Heterogeneity; Hour; Human; Image; Immigration; In Vitro; Integrins; Lateral; Ligands; MDA MB 231; Magnetism; Malignant Neoplasms; Malignant neoplasm of ovary; Mammary Gland Parenchyma; Measurement; Measures; Mechanics; Mediating; Membrane; Mesenchymal; Methodology; Microscope; Migration Assay; Modeling; Mus; Nanotechnology; Neoplasm Metastasis; Ovarian; Ovarian Tissue; Pathogenesis; Patients; Process; Property; Rheology; S-Phase Fraction; Signal Transduction; Signal Transduction Pathway; Specimen; Staging; System; Time; Tissues; Translations; Tumor Biology; Tumorigenicity; abstracting; bioimaging; cancer cell; cancer therapy; cell motility; data acquisition; epithelial to mesenchymal transition; high throughput screening; indexing; inhibin; insight; instrument; magnetic beads; malignant breast neoplasm; member; metastatic process; migration; neoplastic cell; neuronal cell body; prognostic; prototype; receptor; response; software development; tool; treatment effect; tumor; tumor progression

DESCRIPTION (provided by applicant): Array Microscope Assay for Cancer Cell Mechanics Abstract As cells become cancerous, characteristic changes take place in their behavior that affect cell division as well as the ability of the cell to migrate or metastasize. Metastatic behavior, including cell migration, motility and adhesion, is one of the most damaging hallmarks of cancer. Current assays of cell metastases involve the observation of the lateral mobility of cells in a "scratch" assay, or the translation of cells through porous membranes. These assays usually take several hours to days of cell tracking. Metastatic potential has recently been associated with protrusive ability and cell body mechanical properties. We propose to replace the migration assay with one that measures the cell stiffness and cell mechanical response. This involves performing a calibrated tug on the cell with the measurement of the probe displacement. This measurement takes only seconds. This would allow the replacement of a five to forty eight hour assay with a one minute assay. More important than the simple benefit of a faster measurement on a single specimen, we propose an assay system that will allow high throughput methodologies to be applied to elucidating the time course of the biochemical pathways at the heart of the mechanical, and hence, metastatic propensity. We currently have a prototype multiwell assay system demonstrated on cancer cell mechanics. Our next steps are to move from a 16 well prototype to a 96 well assay, and to validate our system on cell lines and on ex-vivo tumor cells. Our development of high throughput force assays will be applied to relate tumorigenicity to the regulated expression of TGF-2 superfamily receptors and subsequent TGF-2 superfamily signaling. TGF-2 and the related TGF-2 superfamily ligands, the bone morphogenetic proteins (BMPs) and inhibin, are potent regulators of normal epithelial cell proliferation, differentiation, survival and migration, with frequent disruption in these homeostatic mechanisms resulting in human cancers and driving human cancer progression, including the metastatic process. We will assess dynamic changes in biomechanical properties during epithelial- mesenchymal transition (EMT), and investigate the migratory, invasive and metastatic potential of these cell models both in vitro (cell lines) and ex vivo and correlate these results with the biomechanical measurements. These measurements will validate our high throughput force system for a wide variety of cancer cell biology studies, enabling the elucidation of the biochemical and genetic determinants of metastatic behavior. PUBLIC HEALTH RELEVANCE: Array Microscope Assay for Cancer Cell Mechanics Narrative We will develop a high throughput force assay system validated on cancer cell lines and on ex-vivo tumor specimens. This powerful system will be ready to be used for discovery of biochemical and genetic determinants of cancer cell metastatic properties to better understand the basic science, diagnosis and treatment of cancer.

描述（由申请人提供）：癌细胞力学的阵列显微镜分析 摘要当细胞癌变时，其行为会发生特征性变化，影响细胞分裂以及细胞迁移或转移的能力。转移行为，包括细胞迁移、运动和粘附，是癌症最具破坏性的标志之一。目前的细胞转移分析涉及在“划痕”分析中观察细胞的横向移动性，或细胞通过多孔膜的平移。这些测定通常需要几个小时到几天的细胞追踪。最近，转移潜能与突出能力和细胞体机械特性相关。我们建议用测量细胞刚度和细胞机械响应的迁移测定法代替迁移测定法。这涉及通过测量探头位移对电池进行校准拖拉。该测量仅需几秒钟。这将允许用一分钟的测定代替五到四十八小时的测定。比对单个样本进行更快测量的简单好处更重要的是，我们提出了一种分析系统，该系统将允许应用高通量方法来阐明机械核心的生化途径的时间进程，从而阐明转移倾向。我们目前拥有一个在癌细胞力学上展示的原型多孔测定系统。我们的下一步是从 16 孔原型转向 96 孔测定，并在细胞系和离体肿瘤细胞上验证我们的系统。我们开发的高通量力测定将用于将致瘤性与 TGF-2 超家族受体的调节表达以及随后的 TGF-2 超家族信号传导联系起来。 TGF-2 和相关的 TGF-2 超家族配体、骨形态发生蛋白 (BMP) 和抑制素是正常上皮细胞增殖、分化、存活和迁移的有效调节剂，这些稳态机制的频繁破坏导致人类癌症并驱动人类癌症进展，包括转移过程。我们将评估上皮-间质转化（EMT）过程中生物力学特性的动态变化，并研究这些细胞模型在体外（细胞系）和离体的迁移、侵袭和转移潜力，并将这些结果与生物力学测量结果相关联。这些测量将验证我们的高通量力系统适用于各种癌细胞生物学研究，从而阐明转移行为的生化和遗传决定因素。 公共卫生相关性：癌细胞力学叙述的阵列显微镜分析 我们将开发一种高通量力分析系统，并在癌细胞系和离体肿瘤样本上进行验证。这个强大的系统将准备好用于发现癌细胞转移特性的生化和遗传决定因素，以更好地了解癌症的基础科学、诊断和治疗。