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.

摘要当细胞发生癌变时，它们的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。目前的细胞转移检测包括观察细胞在“划痕”试验中的横向移动，或细胞通过多孔膜的翻译。这些检测通常需要数小时到数天的细胞追踪。转移潜能最近与突出能力和细胞体力学特性有关。我们建议用一种测量细胞刚度和细胞力学反应的方法来代替迁移试验。这包括在测量探针位移的情况下对电池进行校准拖曳。这个测量只需要几秒钟。这将允许用一分钟的测定代替5到48小时的测定。比单个标本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，该系统将允许高通量方法应用于阐明机械和转移倾向核心的生化途径的时间过程。我们目前有一个多孔实验系统的原型，展示了癌细胞力学。下一步，我们将从16孔的原型实验发展到96孔的实验，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分析将用于将致瘤性与TGF-2超家族受体的调控表达和随后的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（BMPs）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，这些稳态机制经常被破坏，导致人类癌症和驱动人类癌症进展，包括转移过程。我们将评估上皮-间充质转化（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）和离体中的迁移、侵袭和转移潜力，并将这些结果与生物力学测量结果联系起来。这些测量将验证我们的高通量力系统用于各种癌细胞生物学研究，使转移行为的生化和遗传决定因素得以阐明。