Abstract
BACKGROUND: Ammonia metabolism plays a crucial role in cancer progression and immune modulation. However, the prognostic relevance and immunological effects of ammonia-induced cell death (ACD) in HNSC remain insufficiently studied. ACD is a newly characterized form of regulated cell death caused by pathological ammonia accumulation, leading to mitochondrial dysfunction, oxidative stress, and DNA damage. METHODS: A prognostic model was developed using random survival forests (RSF), with performance evaluated through concordance index (C-index) and survival analysis. Immune landscape profiling was performed using ssGSEA and Tracking Tumor Immunophenotype (TIP) analysis. Single-cell transcriptomic data were also used to examine ACD-associated gene expression in distinct cellular populations. RESULTS: KM survival analysis revealed that a high ammonia-death signature (ADS) was significantly associated with worse survival outcomes. Multivariate Cox regression identified ADS as an independent predictor of prognosis. Immune profiling showed that high ADS correlated with an immunosuppressive microenvironment, reduced immune infiltration, and impaired immune function. Glutaminase (GLS) was identified as a key ACD-related gene with high diagnostic accuracy and prognostic relevance. Single-cell RNA sequencing analysis demonstrated elevated GLS expression in malignant and stromal cells, with cell-cell communication analysis suggesting its involvement in immune evasion and metabolic reprogramming. CONCLUSIONS: This study establishes ADS as a novel prognostic biomarker in HNSC, linking ammonia metabolism with tumor progression and immune suppression. These findings highlight potential therapeutic targets for metabolic and immunotherapeutic interventions in HNSC.