Unraveling the Molecular Mechanisms of Glioma Recurrence: A Study Integrating Single‐Cell and Spatial Transcriptomics

OBJECTIVE: Glioma recurrence severely impacts patient prognosis, with current treatments showing limited efficacy. Traditional methods struggle to analyze recurrence mechanisms due to challenges in assessing tumor heterogeneity, spatial dynamics, and gene networks. Single-cell combined spatial transcriptomics (ST) offers innovative solutions. METHODS: We analyzed glioma mRNA data from TCGA and single-cell and ST data from GEO. Following quality control, dimensionality reduction, clustering, and cell annotation of single-cell sequencing data, we identified cell types exhibiting significantly aberrant distributions between primary and recurrent samples by analyzing the deviation degree of Ro/e values. Fibroblasts demonstrating the greatest intergroup differences were subsequently selected as the key cellular population for further investigation. Key differentially expressed genes (DEGs) were identified via random survival forest analysis. Drug sensitivity was assessed using GDSC. Deconvolution algorithms mapped cellular spatial distribution, while PROGENy quantified pathway activity. MISTy modeling revealed cell-cell interactions. RESULTS: Fibroblasts were the primary recurrence-associated subpopulation, with marker genes enriched in extracellular matrix and adhesion pathways. AEBP1, ZNF708, and TSHZ2 were identified as key genes: AEBP1/TSHZ2 correlated with poor prognosis, while ZNF708 showed an inverse trend. These genes were linked to chemosensitivity (Irinotecan, Carmustine, Vincristine, and Cisplatin). Recurrent tumors exhibited increased plasma cell infiltration, with key genes regulating IL-17, Notch, and Toll-like receptor pathways. Spatial analysis highlighted oligodendrocyte-astrocyte interactions in the tumor microenvironment. INTERPRETATION: Fibroblasts drive glioma recurrence, with AEBP1, ZNF708, and TSHZ2 predicting recurrence and chemoresistance. These genes promote immune suppression (via plasma cells) and activate recurrence pathways. Oligodendrocyte-astrocyte interactions shape the recurrent microenvironment, suggesting new therapeutic targets.