Array Microscope Assay for Cancer Cell Mechanics

描述（由申请人提供）：癌细胞力学的阵列显微镜分析摘要随着细胞癌变，其行为发生特征性变化，这些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。目前细胞转移的测定涉及在“划痕”测定中观察细胞的侧向移动性，或细胞通过多孔膜的平移。这些检测通常需要几个小时到几天的细胞跟踪。最近转移潜力与突出能力和细胞体机械性能有关。我们建议用一种测量细胞刚度和细胞机械响应的方法代替迁移测定。这涉及在测量探针位移的情况下对单元执行校准的拖拽。这个测量只需要几秒钟。这将允许用一分钟测定代替五至四十八小时测定。更重要的是比一个简单的好处，更快的测量一个单一的标本，我们提出了一个测定系统，将允许高通量的方法被应用于阐明的时间过程中的生化途径的心脏的机械，因此，转移倾向。我们目前有一个原型多孔检测系统上展示癌细胞力学。我们的下一步是从16孔原型转移到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应用于将致瘤性与TGF-2超家族受体的调节表达和随后的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（BMP）和TGF-2受体，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用于各种癌细胞生物学研究，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叙述我们将开发一种高通量的力分析系统，在癌细胞系和离体肿瘤标本上进行验证。这个强大的系统将准备用于发现癌细胞转移特性的生物化学和遗传决定因素，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.