Abstract
Extensive studies have demonstrated the relationship between metabolic reprogramming and the tumor microenvironment. Here, we characterized the head and neck squamous cell carcinoma (HNSCC) evolutionary landscape using spatial metabolomics/transcriptomics, single-cell transcriptomics, and bulk multi-omics. Metabolic heterogeneity during HNSCC malignant transformation was identified, with significant enrichment in the purine metabolism. Integrating single-cell and bulk data, we developed a robust ligand-receptor-based signature (LRS) linked to NT5E, a key upstream regulator of purine metabolism, which served as an independent prognostic indicator. The low LRS subtype was associated with a high proportion of immune cell infiltration and improved response to immunotherapy. Notably, in vitro and in vivo experiments demonstrated that AMIGO2, a core molecule within the LRS, regulates tumor-associated purine metabolism, and that its downregulation suppresses tumor cell invasion and migration, inhibits myofibroblast differentiation, and promotes immune effector cell infiltration. Moreover, combining AMIGO2 targeting with anti-PD-1 therapy yielded superior efficacy. Consistent validation was also obtained in a clinical cohort of HNSCC and premalignancy patients.