A cell death program-based tumor signature stratifies prognosis, immune landscape, and therapeutic response in glioma

Glioma exhibits profound intratumoral heterogeneity driven by coordinated cell death programs and dynamic tumor-microenvironment interactions. However, the transcriptional landscape linking inflammatory cell death signatures with malignant cell states, genomic alterations, and therapeutic vulnerability remains incompletely defined. We sought to develop a prognostic cell death-associated signature and elucidate its biological and clinical relevance in glioma.

Using transcriptomic and clinical data from TCGA-GBMLGG and external validation cohorts, we constructed a pyroptosis-apoptosis-associated gene signature (PA.Sig) through integrative survival modeling. Genomic alteration profiling, pathway enrichment analyses, and tumor immune microenvironment characterization were performed. Single-cell RNA sequencing datasets were analyzed to delineate malignant cell states and intercellular communication patterns. Functional validation was conducted in glioma cell lines treated with chemotherapeutic agents.

PA.Sig stratified patients into high- and low-risk groups with significantly different overall survival and independent prognostic value. High-risk tumors were characterized by increased somatic mutation burden, enrichment of cell cycle and inflammatory pathways, and a malignant-centered outgoing signaling architecture. Single-cell analyses revealed preferential enrichment of MES-like and AC/MES-like states in high-risk tumors, accompanied by intensified ligand-receptor interactions with immune and stromal compartments. Despite increased immune infiltration, high-risk tumors displayed features of immune dysfunction and exhaustion. In vitro, chemotherapy induced concomitant apoptotic and pyroptotic morphologies in glioma cells, supporting coexistence of lytic and non-lytic death programs. Collectively, PA.Sig captured a coordinated inflammatory cell death landscape associated with aggressive cellular states and immune remodeling.

Our study defines a clinically relevant inflammatory cell death signature that integrates tumor cell state plasticity, genomic instability, and immune microenvironment dynamics in glioma. PA.Sig may serve as a prognostic biomarker and provide a rationale for combinatorial strategies targeting cell death pathways and immune modulation.