High‐Throughput Cellular Heterogeneity Analysis in Cell Migration at the Single‐Cell Level

Abstract

cancer metastasis is a complicated multistep process, migration and intravasation of tumor cells from the tumor stroma to a capillary bed or lymphatic vessel represent critical early steps. [3] In addition, considerable evidence suggests that cancer metastasis can be driven by epithelialto-mesenchymal transition (EMT), [4][5][6] a developmental program in which epithelial cells acquire migratory and invasive phenotypes. [7][8][9][10] Inhibition or activation of EMT significantly limits or increases metastasis, respectively. [11][12][13] To elucidate the correlation between cancer migration and metastasis, we previously developed a high-throughput microfluidic platform that can measure the individual migratory capability of thousands of cancer cells and selectively isolate fast-moving subpopulations. [14,15] As expected, we found that highly migratory breast cancer cells metastasized more than non-migratory cells. [15] Furthermore, through whole transcriptome sequencing of migratory and control cancer cells, differentially expressed genes that correlate with clinical outcomes in breast cancer were identified. [16] The distinct gene expression profile of migratory cancer cells highlights the possibility to selectively inhibit this metastatic sub-population. As migratory breast Cancer cell migration represents an essential step toward metastasis and cancer deaths. However, conventional drug discovery focuses on cytotoxic and growthinhibiting compounds rather than inhibitors of migration. Drug screening assays generally measure the average response of many cells, masking distinct cell populations that drive metastasis and resist treatments. Here, this work presents a high-throughput microfluidic cell migration platform that coordinates robotic liquid handling and computer vision for rapidly quantifying individual cellular motility. Using this innovative technology, 172 compounds were tested and a surprisingly low correlation between migration and growth inhibition was found. Notably, many compounds were found to inhibit migration of most cells while leaving fast-moving subpopulations unaffected. This work further pinpoints synergistic drug combinations, including Bortezomib and Danirixin, to stop fast-moving cells. To explain the observed cell behaviors, single-cell morphological and molecular analysis were performed. These studies establish a novel technology to identify promising migration inhibitors for cancer treatment and relevant applications.

Microfluidic single-cell migration chip reveals insights into the impact of extracellular matrices on cell movement

“…1A). 59 Initially, cells were loaded into the upper wells, followed by the introduction of chemoattractant into the lower wells after cell adhesion, thereby stimulating cell chemotaxis toward the channels (Fig. 1B and C).…”

“…In order to analyze a large number of cell migration images, we previously developed an automated image processing toolkit using MATLAB 2022b, which includes features for image registration and quality control. 59 The image registration process involves utilizing two characteristics of the images: 1 the blue auto-fluorescence of Silicone on the ProPlate® 64-well device to identify the region of cell migration channel, and 2 the brightfield image of migration channels to determine rotation angles and separate individual channels. As the patterns of migration channels extend beyond the core cell migration area, these patterns can be identified using brightfield imaging.…”

“…37–58 Recently, we have developed a high-throughput microfluidic migration platform that utilizes robotic liquid handling and computer vision to rapidly quantify individual cellular motility. 59…”

“…Recently, we have developed a high-throughput microfluidic migration platform that utilizes robotic liquid handling and computer vision to rapidly quantify individual cellular motility. 59 With the cutting-edge microfluidic platform, we delve into the influence of extracellular matrix (ECM) on breast cancer cellular migration. The ECM, a complex network of proteins and carbohydrates enveloping cells, assumes pivotal roles in tumor development and progression.…”

High Throughput Confined Migration Microfluidic Device for Drug Screening

“…Drugs focusing on inhibiting confined migration are rare, the only reported inhibitor work efficiently is the combination of Bortezomib and Danirixin by a very latest work. [34] Those two drugs are screened by a single-cell lateral confined migration device, which can inhibit both cell growth and migration. Similarly, by further validation, we precisely screened out the most significant confined migration inhibitors for pancancer.…”