Integrative immune subtyping of HNSCC reveals clinically relevant phenotypes and treatment-associated transitions

Head and neck squamous cell carcinoma (HNSCC) exhibits profound heterogeneity in clinical presentation, treatment response, and immune landscape. While prior classification systems have identified molecular and immune subtypes in this disease, their applicability to real-world clinical settings remains restricted to small, homogeneous cohorts and limited by lack of multimodal data integration and interpretation. We performed integrated multi-omics analysis including transcriptomic, genomic (copy number, single-nucleotide variants) on 1,149 tumors from 1,102 HNSCC patients across treatment settings. Using the Similarity Network Fusion (SNF) algorithm, we defined immune subtype clusters (ISCs) based on the full immune gene landscape. These clusters were characterized using mutational, transcriptional, and immune cell enrichment analyses, and mapped to hypoxia and traditional subtypes. Associations with clinical outcomes, including progression-free survival, were evaluated across first-line and post-metastatic treatment settings. Four distinct immune subtype clusters (ISC1-ISC4) were identified: ISC1: immune-cold and EMT-enriched; ISC2: immune activated; ISC3: mixed immune-regulatory and stromal-enriched phenotype; and ISC4: immunosuppressed. Distinct treatment response patterns were observed across subtypes in subjects treated with checkpoint inhibitors, chemotherapy, and combination regimens. 44 Patients with matched pre/post treatment tumors revealed treatment-associated transitions between immune subtypes: checkpoint inhibitor treatment enriched for immune activation, while chemotherapy treatment enriched for immunosuppressive signaling pathways. This study provides a clinically relevant immune subtyping framework for HNSCC based on real-world, multi-omics data. These subtypes reflect dynamic tumor-immune states and associated with treatment response and survival, supporting their use in guiding immune-based therapy in HNSCC.