Array Microscope Assay for Cancer Cell Mechanics

描述(申请人提供)：癌症细胞力学的阵列显微镜分析摘要当细胞癌变时，它们的行为会发生特征变化，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、运动和粘连，是癌症最具破坏性的特征之一。目前的细胞转移检测方法包括观察细胞在“划痕”实验中的横向移动性，或细胞通过多孔膜的转移。这些检测通常需要几个小时到几天的细胞跟踪。转移潜能最近被认为与突起能力和细胞体的机械性能有关。我们建议用一种测量细胞硬度和细胞机械反应的方法来代替迁移实验。这包括在测量探头位移的情况下对电池执行校准的拖拽。此测量只需几秒钟。这将允许用一分钟的化验取代5到48小时的化验。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，该系统将允许高通量方法被应用于阐明位于机械转移倾向中心的生化途径的时间进程，从而阐明转移倾向。我们目前有一个原型多孔分析系统演示了癌细胞的力学。我们的下一步是从16孔的原型转移到96孔的测试，并在细胞系和体外肿瘤细胞上验证我们的系统。我们开发的高通量测力分析将被应用于将肿瘤发生与转化生长因子-2超家族受体的调节表达以及随后的转化生长因子-2超家族信号联系起来。转化生长因子-2及相关的转化生长因子-2超家族配体，骨形态发生蛋白和抑制素，是正常上皮细胞增殖、分化、存活和迁移的有效调节因子，这些动态平衡机制经常被破坏，导致人类癌症，并推动人类癌症的进展，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)和体外的迁移、侵袭和转移潜力，并将这些结果与生物力学测量相关联。这些测量将验证我们用于多种癌细胞生物学研究的高通量FORCE系统，从而能够阐明转移行为的生化和遗传决定因素。 公共卫生相关性：癌细胞力学的阵列显微镜分析我们将开发一种在癌细胞系和体外肿瘤标本上验证的高通量测力系统。这一强大的系统将被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。

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.