Immunoinformatics-driven design of a multiepitope vaccine against the EGFR extracellular domain for head and neck squamous cell carcinoma.

Epidermal growth factor receptor (EGFR) is overexpressed in more than 90% of head and neck squamous cell carcinoma (HNSCC) cases and plays a critical role in tumor progression, metastasis, and therapeutic resistance, making it an attractive target for immunotherapy. This study aimed to design and comprehensively evaluate a multi-epitope peptide vaccine targeting the extracellular domain of EGFR using an integrated immunoinformatics approach. Linear B-cell, helper T lymphocyte (HTL), and cytotoxic T lymphocyte (CTL) epitopes were predicted and screened for antigenicity, immunogenicity, allergenicity, toxicity, HLA binding affinity, population coverage, and IFN-γ induction. Selected epitopes were assembled into a multi-epitope vaccine construct and evaluated through structural modeling, molecular docking, molecular dynamics simulation, immune simulation, and codon optimization. Epitope conservancy was assessed across reviewed EGFR isoforms, followed by structural and HNSCC somatic mutation mapping. The vaccine construct exhibited favorable antigenic, non-allergenic, and non-toxic properties with broad predicted population coverage. Among the HTL epitopes, CQGTSNKLTQLGTFE was predicted to induce IFN-γ. Molecular docking and dynamics simulations demonstrated stable receptor interactions, whereas immune simulation predicted robust humoral and cellular immune responses. Six of the ten selected epitopes were completely conserved across all reviewed EGFR isoforms, while the remaining epitopes retained 75% conservancy. Importantly, none of the recurrent extracellular EGFR mutations identified in 619 profiled HNSCC tumors overlapped with the selected epitopes. These findings support the proposed EGFR-targeted multi-epitope vaccine as a promising immunotherapeutic candidate for HNSCC and provide a strong computational foundation for future experimental validation.