High-throughput intracellular microrheology: a new tool for cancer research

总结 癌症的死亡率和发病率与肿瘤的侵袭和转移密切相关， 分子机制知之甚少。在更好地揭示其病因之前， 新的癌症疗法仍将是经验性的。细胞运动，这驱动癌症转移，涉及 细胞骨架的动态和受调节的重排。我们和其他几个小组的工作 已经表明，细胞骨架表型通常伴随着细胞骨架的急剧变化。 细胞骨架的粘弹性，这反过来又调节细胞骨架的能力， 在前缘产生净推力并允许细胞改变其形状。 细胞机械性质的变化长期以来被预测与转移性肿瘤相关。 潜力然而，当前的细胞力学方法具有严重的缺点-包括细胞生长时间。 测量，缺乏多路复用，测量的模糊性-这阻止了直接测试 重要的假设。本研究的目的是：开发一种高度优化的高通量 弹道注射纳米流变学（htBIN）技术平台，用于测量微机械性能 在癌细胞中快速（<30 s/细胞）和可靠地，并评估这些生物物理特性作为一种 通过比较低侵袭性和高侵袭性卵巢癌细胞的细胞迁移和侵袭功能 这些细胞都来自约翰霍普金斯医院的病人。拟议的文书， 该方法基于多颗粒微观流变学，与当前的方法相比具有关键优势， 细胞力学我们的设备将作为癌症研究的新工具，研究细胞力学， 癌细胞迁移和粘附的背景，并可能最终作为患者的诊断工具 卵巢癌的高危人群，补充更传统的生物分子标记物， 临床环境中的癌症 虽然我们提出的细胞力学方法是先验适用于检测细胞内 任何类型的癌细胞的机械差异，这个项目的主要重点是卵巢癌。 卵巢癌被选为本研究中的疾病模型，因为它代表了最常见的疾病之一。 女性的侵袭性癌症

Summary Cancer mortality and morbidity are critically related to tumor invasion and metastasis in which the molecular mechanisms are poorly understood. Until their etiology is better revealed, attempts to develop new cancer therapeutics would remain empirical. Cell motility, which drives cancer metastasis, involves dynamic and regulated re-arrangements of the cytoskeleton. Our work and that of several other groups have shown that cytoskeleton phenotypes are typically accompanied by drastic changes in the viscoelastic properties of the cytoskeleton, which in turn modulate the ability of the cytoskeleton to generate net pushing forces at the leading edge and allow the cell to change its shape. Changes in cell mechanical properties have long been predicted to correlate with metastatic potential. However, current cell-mechanics approaches suffer from serious drawbacks - including time of measurement, lack of multiplexing, ambiguity of measurements - which prevent a direct test of this important hypothesis. The objective of this study is to: develop a highly-optimized high-throughput ballistic injection nanorheology (htBIN) technological platform to measure the micromechanical properties in cancer cells rapidly (< 30 s per cell) and reliably, and to assess these biophysical properties as a function of cell migration and invasion by comparing ovarian cancer cells of low and high invasive nature to normal cells, all obtained from patients at the Johns Hopkins Hospital. The proposed instrument, which is based on multiple-particle microrheology, presents key advantages over current approaches to cell mechanics. Our device will serve as a new tool for cancer research to study cell mechanics in the context of cancer cell migration and adhesion and may ultimately serve as a diagnostic tool for patients who are at high risk for ovarian cancer, complementing more conventional biomolecular markers of cancer in a clinical setting While our proposed approach to cell mechanics is a priori applicable to detect intracellular mechanical differences in any type of cancer cells, a primary focus of this project is ovarian cancer. Ovarian cancer was selected as the disease model in this study because it represents one of the most aggressive cancers in women.