Biophysical Benchmarks of Malignancy in Primary Breast Tumor Cells

由于缺乏对潜在物理原理的机械知识以及三维(3D)侵袭测试系统有限的生理学相关性，对癌细胞在组织中的侵袭过程知之甚少。这项提议将系统地检验原代乳腺肿瘤细胞的机械性能与其转移潜力之间存在机械联系的假设。在2D和3D环境中比较原代患者来源的肿瘤细胞和正常乳腺细胞，我们将确定迁移细胞的硬度、形状、突起动力学、机械转导和牵引力。此外，我们将使用各种不同硬度、多孔性和粘附性的天然细胞外基质来探索细胞迁移和侵袭的统计特性。目前的体外细胞外基质模型往往给出相互矛盾的结果，远远不能复制体内的生理状况。因此，我们将开发具有可调和精确可控参数的3D细胞培养系统，使我们能够使用高分辨率成像和新的高通量方法来监测原代乳腺癌细胞的侵袭。我们预测已转移的原发肿瘤细胞与未转移的原发肿瘤细胞在生物力学特性和侵袭行为方面存在显著差异。这些体外预测将与同一患者队列的体内临床结果相关联。

The process of cancer cell invasion in tissue is poorly understood due to a lack of mechanistic knowledge of underlying physical principles and limited physiological relevance of 3-dimensional (3D) invasion test systems. This proposal will systematically test the hypothesis of a mechanistic link between mechanical properties of primary breast tumor cells and their metastatic potential. Comparing primary patient-derived tumor and normal breast cells in 2D and 3D environments, we will determine the stiffness, shape, protrusion dynamics, mechanotransduction and traction forces of migrating cells. Moreover, we will explore the statistical properties of cell migration and invasion, using a variety of differently stiff, porous and adhesive natural extracellular matrices. Current in vitro extracellular matrix models often give contradictory results and are far from replicating the physiological situation in vivo. Therefore, we will develop 3D cell culture systems with tunable and precisely controllable parameters that will allow us to monitor the invasion of primary breast cancer cells with high-resolution imaging and novel high-throughput methods. We predict significant differences in the biomechanical properties and the invasion behavior between primary tumor cells that have metastasized versus those that have not. These in vitro predictions will be correlated with in vivo clinical findings from the same patient cohort.