ANXA2+/TNR- glioblastoma subpopulation drives tumor progression through via Src-STAT3 axis and neutrophil remodeling.

BACKGROUND

Glioblastoma (GBM) exhibits remarkable cellular heterogeneity, but the functional roles of distinct tumor subpopulations remain incompletely understood.

METHODS

We employed single-cell RNA sequencing, flow cytometry, and in vitro functional assays to identify and characterize an ANXA2+/TNR- subpopulation in GBM. The stem-like phenotype, key signaling pathways, and interactions with neutrophils were evaluated using limiting dilution assays, 3D culture, co-culture systems, and transcriptomic profiling. Functional relevance was validated by pharmacological inhibition and xenograft models.

RESULTS

ANXA2+/TNR- GBM subpopulation with prominent stem-like features characterized by enhanced self-renewal and tumor-initiating capacity. Functional assays revealed that ANXA2 enhanced GBM cell proliferation, migration, and 3D growth while repressing TNR expression, correlating with aggressive clinicopathological features. Mechanistically, ANXA2 activated the Src-STAT3 signaling cascade, which reinforced stemness-associated transcriptional programs and sustained malignant progression. Additionally, ANXA2 activated the Src-STAT3 pathway, inducing a STAT3-dependent secretory program characterized by robust IL-8 release. In concert with tumor-derived TGF-β, ANXA2-driven IL-8 polarized neutrophils toward an ELANE+/N2 immunosuppressive phenotype, which further secreted MMP2/9 to amplify tumor invasion and growth. Pharmacological inhibition of Src or STAT3 effectively impaired stemness maintenance, suppressed neutrophil N2 polarization, and attenuated GBM growth in xenograft models.

CONCLUSIONS

Our findings identify ANXA2+/TNR- cells as a critical driver of GBM stemness and immune evasion through the Src-STAT3 axis and neutrophil N2 polarization. Targeting this subpopulation and its signaling network may provide new therapeutic strategies for overcoming GBM aggressiveness.