Tumor Associated Neutrophils Extend NETs To Spatially Drive Necroptosis In Glioblastoma

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Abstract

BackgroundGlioblastoma is pathologically characterized by extensive pseudopalisading necrosis, a hallmark linked to therapeutic resistance and dismal prognosis. While traditionally viewed as a passive ischemic event, the precise radiogenomic drivers and spatial immune architectures orchestrating active tumor necrosis remain poorly understood. MethodsWe integrated macroscopic clinical MRI radiomics with public genomic datasets (n=3,639), single-cell spatial transcriptomics, and high-resolution protein immunofluorescence (IF) assays in a novel co-mutant murine model to map the necrotic microenvironment. ResultsRadiomic analysis of patient MRIs demonstrated that high tumoral necrotic volume serves as a powerful independent biomarker for poor long-term survival. This macro-structural necrosis phenotype is radiogenomically driven by cell-intrinsic TP53, PTEN, and NF1 (PPN) co-mutations. Spatial transcriptomic profiling of PPN-deficient tumors revealed distinct cellular topography, separating a proliferative, invasive niche (EGFR+ SOX+, TypeInv) from a hyper-aggressive necrotic niche (ERBB3+, TypeNec). Mechanistically, TypeNec cells deploy a prominent CCL5/CXCL2 chemokine milieu that actively recruits and prompts tumor-associated neutrophils to undergo neutrophil extracellular trap (NET) formation. This overactive neutrophil infiltrate drives a feed-forward loop of intrinsic tumor necroptosis mediated by the release of the damage-associated molecular pattern (DAMP) protein HMGB2. This cascade could be therapeutically dismantled using targeted anti-inflammatory compounds. Finally, multi-cohort clinical validation confirmed that an HMGB2high signature dictates a high-risk, myeloid-rich immunosuppressive microenvironment in glioblastoma patients. ConclusionsOur findings redefine glioblastoma necrosis as an active, mutation-driven immunopathological process. The HMGB2-mediated DAMP necroptosis cascade and associated myeloid chemokines provide robust prognostic biomarkers and immediate therapeutic vulnerabilities to disrupt the immunosuppressive necrotic niche. Key PointsO_LIClinical Stratification via Radiomics: Preoperative MRI analysis of 1,079 glioblastoma patients establishes a necrotic core volume threshold of 18.57% as a robust, non-invasive biomarker for early survival divergence and dismal long-term prognosis. C_LIO_LIGenomic Orchestration of Necrosis: Cross-species modeling identifies a high-risk somatic PPN co-mutation profile yielding two topographically distinct niches: an invasive niche EGFR+ SOX+, TypeInv) from a hyper-aggressive necrotic niche (ERBB3+, TypeNec). C_LIO_LIVascular Loop and DAMP Cascade: TypeNec cells deploy a CCL5/CXCL2 chemokine milieu driving perivascular neutrophil recruitment and MPO+CitH3+ NETosis, sparking a feed-forward loop of hypoxia and necrosis driven by tumor-derived HMGB2high that is pharmacologically targetable. C_LI Online Key Points (85 characters)O_LIPreoperative MRI biomarker of necrotic threshold determines long-term prognosis. C_LIO_LICross-species co-mutation profile yielding distinct invasive and necrotic niches. C_LIO_LINeutrophil extracellular traps spark DAMP-targetable feed-forward necroptosis loop. C_LI Importance of the StudyTumor necrosis is a hallmark diagnostic feature of glioblastoma, historically characterized as a passive consequence of local tissue ischemia. This study redefines glioblastoma necrosis as an active, genetically driven immunopathological process. By integrating macroscopic clinical MRI radiomics from over 1,000 patients with single-cell spatial transcriptomics and a novel co-mutant mouse model, we reveal that cell-intrinsic TP53, PTEN, and NF1 co-mutations actively dictate a hyper-aggressive necrotic niche. Mechanistically, these mutated tumor cells deploy a CCL5/CXCL2 chemokine cascade to recruit neutrophils, triggering an overactive neutrophil extracellular trap (NET) and a HMGB2-mediated damage-associated molecular pattern (DAMP) loop that accelerates tissue destruction and myeloid immunosuppression. Crucially, we demonstrate that this pathogenic axis can be tracked non-invasively via standard neuroimaging and pharmacologically dismantled with anti-inflammatory small molecules, providing immediate diagnostic stratifications and a brand-new therapeutic vulnerability to disrupt the glioblastoma microenvironment. Lay SummaryDead tissue zones inside glioblastoma brain tumors, known as necrosis, are tied to poor patient survival. While long believed to be an accidental result of rapid tumor growth cutting off oxygen, this study reveals that tumor cells actively trigger this cell death. Driven by specific genetic mutations, these cells release distress signals that recruit immune cells called neutrophils. These neutrophils create a destructive, inflammatory loop that worsens the disease. By combining patient MRI scans with advanced spatial tissue mapping, we show that this destructive process can be tracked non-invasively and stopped using anti-inflammatory drugs, offering a new treatment strategy.

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