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Molecular Profiling of Head and Neck Squamous Cell Carcinomas

This study performed targeted DNA sequencing of key NF-κB signaling pathway regulatory genes in matched tumor and normal tissues from patients with HPV-associated head and neck squamous cell carcinoma (HNSCC). A custom Ion Torrent AmpliSeq panel was designed to interrogate ten genes whose products regulate NF-κB activity and are frequently altered in the NF-κB-active subtype of HPV+ HNSCC: TRAF3, CYLD, TRAF2, NFKBIA, TNFAIP3, TRAF6, BIRC2, BIRC3, MYD88, and MAP3K14. The panel comprised 250 overlapping amplicons in a 2-primer pool format, achieving 93.65% coverage across the 24.86 kb target region. Genomic DNA was isolated from formalin-fixed paraffin-embedded (FFPE) macro-dissected tumor and matched normal tissues at the UNC Lineberger Comprehensive Cancer Center Translational Genomics Lab. This sequencing was performed as part of a study developing a clinical assay to identify HPV+ HNSCC subtypes defined by NF-κB activity for the purpose of risk stratification and de-escalation of radiation therapy.