Identification and evaluation of tumor pyroptosis-associated antigens for design a vaccine candidate against lung cancer.

Lung cancer is the leading cause of cancer-related mortality worldwide, emphasizing the need for innovative therapeutic strategies. This study utilized immunoinformatics and structural bioinformatics approaches to design and evaluate a multi-epitope vaccine targeting pyroptosis-associated antigens (CARD8, NAIP, NLRP1, and NLRP3), which are implicated in lung cancer immunology. Fifteen T-cell and B-cell epitopes were identified, analyzed for their antigenicity, non-toxicity, non-allergenicity, and immune-stimulatory potential, and optimized to construct a vaccine with suitable adjuvants and linkers. The vaccine demonstrated high antigenicity, solubility, and stability, as validated through physicochemical analyses. Its three-dimensional structure was modeled, refined, and validated using molecular modeling approaches. Molecular docking studies revealed stable and strong interactions between the vaccine and key immune receptors (TLR2, TLR4, TLR5, TLR3, TLR7, and TLR8), indicating its potential to activate both innate and adaptive immunity. Molecular dynamics simulations confirmed the vaccine's structural stability and solvent accessibility over 100 ns across ten replicas. Immune simulations demonstrated strong immunogenic responses, including elevated antibody titers, memory cell populations, and cytokine production. Codon optimization and in silico cloning further ensured efficient expression in Escherichia coli, facilitating experimental application. These findings underscore the vaccine's promise as a therapeutic candidate against lung cancer, warranting further in vitro and in vivo investigations.