Aggressive Glioblastoma Cells Enhance the Migratory Persistence and Velocity of Less Aggressive Cells to Promote Tumor Dissemination

ABSTRACT Glioblastoma multiforme (GBM) is a highly lethal brain cancer driven by aggressive invasion. Epidermal growth factor receptor (EGFR) is frequently amplified in GBM, with the EGFRvIII mutant enhancing the infiltrative features of EGFRwt‐overexpressing cells. Previously, we identified Src as a key cell‐cell communication mediator between EGFRvIII and EGFRwt cells. However, a quantitative biophysical characterization of how EGFRvIII induces this increased infiltration, specifically detailing altered movement parameters and the underlying mechanisms, remains lacking. Using bulk cell culture and 2‐cell microfluidic chips, we quantitatively analyzed the motility of EGFRwt‐overexpressing and EGFRvIII‐expressing GBM cells. We observed that EGFRvIII cells exhibit higher migration velocity and persistence compared to EGFRwt cells, with a highly persistent subpopulation contributing significantly to these differences. Notably, the co‐culture of EGFRvIII cells enhanced the velocity and migration persistence of EGFRwt‐overexpressing cells, leading to increased spreading. Our findings revealed that Src‐mediated cell‐cell communication from EGFRvIII‐expressing to EGFRwt‐overexpressing cells promotes aggressive GBM spreading by increasing both their velocity and migration persistence at the micro‐environmental level. Inhibiting the Src pathway with dasatinib reversed this pro‐migratory effect, markedly reducing their migration persistence. These insights refine our understanding of GBM infiltration and highlight Src inhibition as a promising strategy in EGFRvIII‐positive tumors.