Single-cell mapping of TMZ-resistant glioma reveals CXCL2/3–CXCR2–associated neutrophil communication

Temozolomide (TMZ) resistance remains a major obstacle to achieving durable treatment responses in glioblastoma (GBM), yet its microenvironmental associations remain incompletely defined. While most studies have focused on tumor-intrinsic mechanisms, the contribution of the tumor microenvironment (TME) to TMZ resistance is less well characterized. Here, by performing transcriptomic profiling of matched control and TMZ-resistant glioma cells based on previously publised result, we identified a prognostic gene signature representing resistant glioma stem-like cells. Projection of this signature onto large-scale single-cell RNA-seq datasets enabled stratification of tumor cells into distinct transcriptional subpopulations with varying resistance profiles. Resistant clusters exhibited enhanced crosstalk with multiple immune cell types compared with non-resistant clusters, with the strongest signals directed toward microglia/macrophages (MG) and a uniquely enriched CXCL2/3-CXCR2 chemotactic axis engaging neutrophils, as revealed by CellChat analysis. Functional assays confirmed that conditioned media from resistant glioma cells promoted neutrophil migration and reactive oxygen species (ROS) production, effects that were attenuated by CXCR2 inhibition. Together, these results define a resistance-associated gene signature that delineates discrete cellular states within gliomas and uncover a specific tumor-neutrophil communication circuit linked to TMZ resistance, highlighting CXCL2/3-CXCR2 signaling as a potential therapeutic target for overcoming treatment resistance.