Abstract C073: Immunoinformatic design and evaluation of a multi-epitope and multivalent mRNA vaccine targeting NSCLC through MAGE-3, MUC-1, and EGF

Abstract Non-small cell lung cancer (NSCLC) is the most common and deadly type of lung cancer, often growing slower than small cell lung cancer; it accounts for 87% of all lung cancers with a 5-year survival rate of 37% for regional NSCLC and 9% for metastatic NSCLC. Despite its prevalence, there are currently few effective therapeutic interventions beyond supportive care. Vaccine development has emerged as a promising method for NSCLC prevention and treatment. mRNA vaccines have emerged as a promising type of vaccine for increased robustness of immune response. Thus, this study uses immunoinformatic tools to design a safe, efficacious, soluble, and non-toxic multivalent mRNA NSCLC vaccine by assembling antigenic, immunogenic, and non-allergenic B- and T-cell (CTL and HTL) epitopes specifically targeting melanoma-associated antigen 3 (MAGE-3), Mucin-1 (MUC-1) protein, and epidermal growth factor (EGF); MAGE-3 has been found to be expressed in NSCLC affected tissue but not normal tissue, indicating involvement with cancer growth; MUC-1 is over-expressed in all epithelial cancers including NSCLC, making it a good broad-spectrum protein target; EGF is especially vulnerable to somatic mutations closely associated with NSCLC metastasis. Using immunoinformatic methods, 39 CTL, 8 HTL, and 31 B-cell epitopes were successfully identified and conjugated. We ordered the epitopes with 5′ cap, 5′ UTR, Kozak sequence, signal peptide (tPA), stop codon, 3′ UTR, and a 120-nucleotide long poly(A) tail at the C-terminal with linker sequences (AAY, GGGGS, KK). The computational findings (physicochemical, structural (secondary and tertiary), 3D refinement analyses, MD simulations, and docking analyses) corroborated the stability, high quality, and hydrophilicity of the proposed construct. The final mRNA construct had a molecular weight of 131069.91 kDA and was deemed soluble (grand average of hydropathicity of -0.459); a high level of thermostability was also observed with a stability index of 50.49 and an aliphatic index of 57.81. The half-life of the construct is over 10 hours in E. coli in vivo. Molecular docking and immune simulation revealed a high binding affinity (below -600 kcal/mol) of the in silico vaccine with artificial toll-like-receptors (TLR2, TLR3, and TLR9). Immune simulations showed a robust immune response. A slow but consistent increase in immunoglobulins (IgG and IgM) and cytokines (IL-4, IL-10, etc.) was observed. A large increase in memory T-cells and B-cells was noted as well, further corroborating the observed enhanced immune response. Codon optimization showed further success, yielding a GC score above 0.9. This immunoinformatic study presents a viable multivalent NSCLC vaccine candidate, suggesting that this mRNA construct can now be validated experimentally and be compared to other vaccine candidates to assess its possible impact in human health. This in silico vaccine offers a groundbreaking platform for further research and therapeutic intervention for lung cancer. Citation Format: Ridhi Gutta, Sreelasya Polavarapu, Suhani Garg. Immunoinformatic design and evaluation of a multi-epitope and multivalent mRNA vaccine targeting NSCLC through MAGE-3, MUC-1, and EGF [abstract]. In: Proceedings of the AACR Immuno-Oncology Conference (AACR IO): Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2026 Feb 18-21; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2026;14(2 Suppl):Abstract nr C073.