On-Chip Clonal Analysis of Glioma-Stem-Cell Motility and Therapy Resistance

Abstract

Enhanced glioma-stem-cell (GSC) motility and therapy resistance are considered to play key roles in tumor cell dissemination and recurrence. As such, a better understanding of the mechanisms by which these cells disseminate and withstand therapy could lead to more efficacious treatments. Here, we introduce a novel micro-/nanotechnology-enabled chip platform for performing live-cell interrogation of patient-derived GSCs with single-clone resolution. On-chip analysis revealed marked intertumoral differences (>10-fold) in single-clone motility profiles between two populations of GSCs, which correlated well with results from tumor-xenograft experiments and gene-expression analyses. Further chip-based examination of the more-aggressive GSC population revealed pronounced interclonal variations in motility capabilities (up to ∼4-fold) as well as gene-expression profiles at the single-cell level. Chip-supported therapy resistance studies with a chemotherapeutic agent (i.e., temozolomide) and an oligo RNA (anti-miR363) revealed a subpopulation of CD44-high GSCs with strong antiapoptotic behavior as well as enhanced motility capabilities. The living-cell-interrogation chip platform described herein enables thorough and large-scale live monitoring of heterogeneous cancer-cell populations with single-cell resolution, which is not achievable by any other existing technology and thus has the potential to provide new insights into the cellular and molecular mechanisms modulating glioma-stem-cell dissemination and therapy resistance.

Combined elevation of TRIB2 and MAP3K1 indicates poor prognosis and chemoresistance to temozolomide in glioblastoma

“…Increasing evidence has indicated that acquired resistance to chemotherapy and radiation leads to the rapid recurrence and poor prognosis of glioma in patients . According to our results, MAP3K1 was the gene that was most correlated with TRIB2.…”

Guided migration analyses at the single-clone level uncover cellular targets of interest in tumor-associated myeloid-derived suppressor cell populations

“…These cells were plated on microtextured polydimethylsiloxane (PDMS) surfaces ( Fig. 1B), which were fabricated via replica molding from photolithographically fabricated silicon masters, and were designed as an array of parallel ridges and grooves with dimensions that have been previously tested in cancer cell dissemination studies (~2 µm × 2 µm with 2 µm spacing) [10][11][12][13] . MDSC motility was monitored at the single-clone level in real time via time-lapse microscopy.…”

“…While targeting the dissemination-based mechanisms by which MDSCs infiltrate the tumor niche could be a viable alternative strategy against MDSC-driven immunosuppression at the tumor site, our understanding of such mechanisms for MDSCs is limited compared to what we know about the dissemination modalities of cancerous tumor cells. Structurally guided migration has been known to play a key role in the escape of cancerous cells from the primary tumor, as well as in dissemination and metastasis [10][11][12][13][14][15] . Nevertheless, to the best of our knowledge, no study has probed MDSC motility under structurally guided dissemination conditions.…”

Wearable Devices for Single-Cell Sensing and Transfection

“…The intrinsic heterogeneity existed in almost any given cell population is a well-known confounding factor with important biological meanings. To properly address this variable and take the cellular heterogeneity into account in research, many platforms that allow biological samples to be analyzed at the single-cell level have been developed and have increasingly significant roles in the studies of genomics, transcriptomics, epigenomics, and proteomics relevant to different fields [57][58][59]. The heterogeneity of cells in a pathological lesion (e.g., a tumor) is likely to determine the therapeutic outcome and long-term prognosis in patients.…”