Collectively, these findings demonstrate that soluble ENO1 activates the TLR4/ERK/SPHK1/S1P signaling cascade to drive GBM malignancy, thereby establishing an autocrine S1P loop that drives malignant progression in GBM, expanding our understanding of ENO1’s non-glycolytic roles and identifying novel therapeutic targets. The gene discussed is TLR4; the disease is glioblastoma.