High Throughput Mechanical Modulatory Assay for Breast Cancer Drug Testing

动态应力微环境可以调制细胞的生物力学，导致明显不同的 正常细胞和癌变细胞的签名。拟议的研究旨在分析生物物理变化。 乳房细胞在反复作用力下被激发时发生在乳腺细胞中的。在这项提议中，我们计划暴露细胞 到顺序变形和确定更全面的用于癌症诊断的生物力学标志， 预后和治疗。提出的“机械调制信号”是细胞变化的结果。 当它穿过多个狭窄区域时的速度可以假想地预测转移 乳腺癌细胞的潜能和药物反应性。我们之前在原子力显微镜和 微流控芯片显示，乳腺癌细胞比健康的癌细胞更柔软、更具流动性。 此外，癌细胞表现出应变软化行为，而正常细胞表现出应变硬化或更少的应变硬化。 态度软化。我们的研究成果将对乳腺癌生物学和药物产生重大影响 发展，因为它意味着癌细胞离开其原始位置时，可以通过挤压变得更软 通过毛孔到达血管并转移，而正常细胞表现出更多的阻力，因此 它们的迁徙速度会放缓，甚至可能会停止。目标1是开发一种高通量微流控芯片，并 相应的流体和图像处理接口，用于分析单个信号的机械调制特征 当细胞穿过多个狭窄时。正常和癌细胞株以及原代细胞都将被使用。 将通过改变总沟道长度和松弛来探索不同的收缩结构 两个后续收缩区域之间的区域。在成功完成这一阶段的 项目中，我们将实现一种高通量的测试，能够对每个细胞进行大约50,000个细胞的生物力学分析 等一下。生物检测将被用来发现每个入伍细胞是否有唯一的调制信号 类别(非侵入性、中度侵入性和高度侵入性)，可用于区分它们以及如何区分 这些签名与约束体系结构相关。目标2将评估化疗的作用 药物对细胞生物力学特征及其相应的细胞骨架结构的影响。目标2是一个 确定微管蛋白干扰药物对活的乳腺细胞力学的影响的基础研究。 同时使用抗癌微管蛋白稳定剂和不稳定剂及其对生物力学的影响 将确定细胞系和原代细胞的调制特征。这个目标将确定细胞是否机械 在药物治疗中，如果观察到细胞由于周期性变形而软化/僵硬，签名发生了变化 已经改变了，改变到了什么程度。

Dynamic stress microenvironments can modulate the biomechanics of cells resulting in distinctly different signatures for normal and caner cells. The proposed research aims at analyzing the biophysical changes occurring in breast cells when they are excited under repetitive forces. In this proposal, we plan to expose cells to sequential deformations and to identify a more comprehensive biomechanical marker for cancer diagnosis, prognosis, and treatment. The proposed “mechanical modulatory signatures” result from changes in the cell velocity as it traverses through multiple constriction regions and can hypothetically predict the metastatic potential and drug responsiveness of breast cancer cells. Our previous work with atomic force microscopy and microfluidic chips reveal that breast cancer cells are softer and more fluidic than their healthy counterparts. Moreover, cancer cells demonstrate strain-softening behavior while normal cells display strain-stiffening or less softening attitude. Our research outcome will have substantial impact on breast cancer biology and drug development as it implies that cancer cells as they leave their original site can become softer by squeezing through pores to reach to blood vessels and metastasize while normal cells show more resistance and hence their migration slows down and potentially stops. Aim 1 is to develop a high throughput microfluidic chip and the corresponding fluidic and image processing interfaces to analyze the mechanical modulatory signature of single cells as they pass through multiple constrictions. Both normal and cancer cell lines and primary cells will be used. Different constriction architectures will be explored by varying the overall channel length and the relaxation regions between two subsequent constriction regions. Upon successful accomplishment of this phase of the project, we will realize a high throughput assay enabling the biomechanical analysis of about 50,000 cells per minute. The bioassays will be used to discover if there are unique modulatory signatures for each enlisted cell category (non-invasive, moderately invasive, and highly-invasive) that can be used to distinguish them and how these signatures are related to the constriction architecture. Aim 2 will be to assess the role of chemotherapy agents on cell biomechanical signatures and their corresponding cytoskeletal architectures. Aim 2 is a fundamental study defining the impact of microtubulin disrupting drugs on the mechanics of living breast cells. Both anti-cancer microtubulin stabilizer and destabilizer drugs will be used and their effect on biomechanical modulatory signatures of cell lines and primary cells will be determined. This aim will identify if cell mechanical signatures have changed upon drug treatment and if cell softening/stiffening observed due to cyclic deformations have altered and to which degree.