NET-Associated Inflammatory Markers in Head and Neck Squamous Cell Carcinoma: Bioinformatic Prioritization and Exploratory Serum Assessment in Patients Undergoing Radiotherapy

Neutrophil extracellular traps (NETs) and inflammatory mediators have been implicated in head and neck squamous cell carcinoma, but their relationship with systemic inflammatory status during radiotherapy remains incompletely understood. This exploratory integrative study combined a literature-guided network-based bioinformatic prioritization strategy, public transcriptomic interrogation, and a small serum-based clinical cohort evaluated during radiotherapy. Network and ontology analyses identified IL6 and CXCL8 as central inflammatory mediators within a NET-associated condition-specific interaction network. Public transcriptomic analyses showed altered expression patterns of selected NETosis-related genes in tumor tissue. To provide complementary clinical context, circulating NET-associated complexes, interleukin-6 (IL-6), and interleukin-8 (IL-8) were measured in patients with head and neck squamous cell carcinoma undergoing radiotherapy and in healthy controls. Patients showed higher serum levels of NET-associated complexes, IL-6, and IL-8 than controls, whereas no significant differences were observed between the two intra-treatment sampling time points. These findings support the relevance of NET-associated inflammatory markers in HNSCC and indicate that systemic inflammatory alterations can be detected in patients sampled during radiotherapy. Given the exploratory design and limited sample size, these findings should be interpreted cautiously and require validation in larger longitudinal studies.