In silico design of a novel therapeutic multi-epitope vaccine against NSCLC: a reverse vaccinology approach

Non-small cell lung cancer (NSCLC) remains one of the leading causes of cancer-related deaths worldwide, highlighting the need for effective immunotherapeutic strategies. This study focuses on the in silico design and evaluation of a multi-peptide vaccine targeting five key TAAs frequently overexpressed in NSCLC (MAGE-A4, NY-ESO-1, MUC1, SURVIVIN, and HER2) and on assessing the effect of incorporating calreticulin as an immunostimulatory adjuvant. Using immunoinformatics approaches, three distinct vaccine constructs were designed: one incorporating calreticulin adjuvant, another without the adjuvant, and a third consisting of the whole sequences of the TAAs. Epitope prediction was performed using NetMHCpan 4.1 and NetMHCIIpan 4.1, followed by screening for antigenicity, allergenicity, immunogenicity, and toxicity. The predicted novel epitopes were assembled into multi-peptide constructs using appropriate linkers, which were then analyzed for physicochemical characteristics and structural interactions with TLRs and HLA alleles. Molecular docking and dynamics simulations demonstrated a strong interaction between the calreticulin-containing construct and TLR4. Immune simulations using the C-ImmSim platform predicted strong activation of both B and T cell responses. While the in silico results are promising, further in vitro and in vivo studies are required to comprehensively evaluate the vaccine’s safety and therapeutic efficacy.