Na+/Ca2+ exchanger 1 (NCX1) drives migration and vasculogenic mimicry in mesenchymal glioblastoma

Abstract Glioblastoma (GBM) is a highly aggressive brain tumor characterized by diffuse invasion, marked cellular plasticity, and poor prognosis, particularly in the mesenchymal subtype. The Na + /Ca 2+ exchanger 1 (NCX1) has been implicated in glioblastoma motility, but its role in patient-derived mesenchymal GBM and vasculogenic mimicry (VM) remains unclear. Here, we investigated NCX1 function in GBM19, a primary mesenchymal glioblastoma stem cell-enriched culture. GBM19 cells displayed coordinated migration and formed Vascular Endothelial (VE)-cadherin-positive loop-like structures consistent with VM, whereas leading-edge cells showed a more migratory phenotype. NCX1 was enriched at the migratory front and was functionally active in both leading-edge and loop cells, as demonstrated by patch-clamp recordings and pharmacological inhibition with bepridil. NCX1 blockade impaired wound closure, disrupted actin organization, altered cell polarity and directional migration, reduced spheroid invasion, and prevented proper loop formation and closure. In vivo, VM-like tumor structures were positive for NCX1 and VE-cadherin. Transcriptomic analysis further showed a positive correlation between NCX1 and the VM-related regulator E74-like factor 2 (ELF2) specifically in mesenchymal GBM, where high ELF2 expression was associated with worse survival. These findings identify NCX1 as a regulator of migration and vasculogenic mimicry in mesenchymal glioblastoma and support its potential as a therapeutic target in aggressive GBM.