On-chip clonal analysis of glioma stem cell motility and therapy resistance

Daniel Gallego-Perez, Lingqian Chang, Junfeng Shi, Junyu Ma, Sung-Hak Kim, Xi Zhao, Veysi Malkoc, Xinmei Wang, Mutsuko Minata, Kwang Joo Kwak, Yun Wu, Gregory Lafyatis, Wu Lu, Derek J. Hansford, Ichiro Nakano, and Ly James Lee

Enhanced glioma stem cell (GSC) motility and therapy resistance are considered to play key roles in tumor cell dissemination and life-threatening recurrence. As such, a better understanding of the mechanisms by which these cells disseminate and withstand therapy could lead to more efficacious treatments for this devastating disease. Here we introduce a novel micro/nanotechnology-based chip platform for performing live-cell interrogation of patient-derived GSC cultures with single-clone resolution. On-chip analysis revealed marked inter-tumoral differences (> 10-fold) in single-clone motility profiles between two populations of patient-derived GSCs, which correlated well with results from supporting tumor xenograft experiments and gene expression analyses. Further chip-based examination of the more aggressive GSC population revealed pronounced inter-clonal 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 sub-population of CD44-high GSCs with strong anti-apoptotic 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.

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