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

描述(申请人提供)：癌症死亡率和发病率与肿瘤的侵袭和转移密切相关，其分子机制尚不清楚。在它们的病因被更好地揭示之前，开发新的癌症疗法的尝试仍将是经验的。推动癌症转移的细胞运动涉及细胞骨架的动态和受调控的重新排列。我们和其他几个小组的工作表明，细胞骨架的表型通常伴随着细胞骨架粘弹性性质的急剧变化，这反过来又调节了细胞骨架在前沿产生净推力的能力，并允许细胞改变其形状。长期以来，细胞力学性质的改变一直被预测与转移潜能相关。然而，当前的细胞力学方法存在着严重的缺陷--包括测量时间、缺乏多路传输、测量的模糊性--这些都阻碍了对这一重要假设的直接检验。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，以快速(每细胞30秒)且可靠地测量癌细胞的微观力学特性，并通过比较低侵袭性和高侵袭性的卵巢癌细胞与正常细胞(均来自约翰·霍普金斯医院的患者)来评估这些生物物理特性作为细胞迁移和侵袭的函数。该仪器基于多颗粒微观流变学，与目前的细胞力学方法相比具有关键的优势。我们的设备将成为癌症研究的一种新工具，在癌细胞迁移和黏附的背景下研究细胞力学，并最终可能作为卵巢癌高危患者的诊断工具，在临床环境中补充更多传统的癌症生物分子标志物。虽然我们提出的细胞力学方法是一种先验方法，可用于检测任何类型癌细胞的细胞内力学差异，但该项目的主要重点是卵巢癌。在这项研究中，卵巢癌被选为疾病模型，因为它代表了女性最具侵袭性的癌症之一。

DESCRIPTION (provided by applicant): 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 seconds 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.