Abstract C076: Immunoinformatic design and evaluation of a multi-epitope and multivalent mRNA vaccine targeting ovarian cancer through NYESO-1, CT45-1, WT-1, FOLR-1, and LAGE-1

Abstract Ovarian cancer is one of the most common gynecological cancers; it is ranked third after cervical and uterine cancer. The lifetime risk of being diagnosed with ovarian cancer is 1.3%, making it the fifth-leading cause of cancer related death among women. The current treatment standard for ovarian cancer primarily involves surgery and platinum-based chemotherapy; however, over 80% of patients experience recurrence despite therapeutic intervention, marking a need for more innovative methods of treatment. While there is not yet an FDA-approved vaccine for ovarian cancer, vaccine development has emerged as a promising method for 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 an efficacious, soluble, and non-toxic multivalent mRNA ovarian cancer vaccine by assembling antigenic, immunogenic, and non-allergenic B- and T-cell (CTL and HTL) epitopes specifically NYESO-1, CT45-1, WT-1, FOLR-1, and LAGE-1; NYESO-1 is a cancer testis antigen expressed in tumor cells generally associated with increased tumor aggression; CT45-1 is a prognostic biomarker of high-grade serous ovarian cancer with increased expression in late-stage disease; WT-1 is a zinc finger transcription factor correlated with aggressive metastasis; FOLR-1 is highly expressed in high-grade serous carcinoma; LAGE-1 is overexpressed in a significant portion of ovarian cancer tissue but not normal tissue. Using immunoinformatic methods, 15 CTL, 21 HTL, and 21 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 106158.33 kDA and was deemed soluble (grand average of hydropathicity of -0.698); a high level of thermostability was also observed with a stability index of 44.78 and an aliphatic index of 49.26. The half-life of the construct is over 20 hours in yeast in vivo. Molecular docking and immune simulation revealed a high binding affinity (below -800 kcal/mol) of the in silico vaccine with artificial toll-like-receptors (TLR3, TLR4, and TLR9). Immune simulations showed a robust immune response. A consistent increase in immunoglobulins (IgG and IgM) and cytokines (IL-4, IL-10, TNF-alpha, IFN-gamma) was observed. A large increase in memory T-cells and B-cells was noted as well, further corroborating the observed enhanced immune response. This immunoinformatic study presents a viable multivalent ovarian cancer vaccine candidate, suggesting that this mRNA construct can now be validated experimentally for potential clinical use. Citation Format: Ridhi Gutta, Sreelasya Polavarapu, Suhani Garg. Immunoinformatic design and evaluation of a multi-epitope and multivalent mRNA vaccine targeting ovarian cancer through NYESO-1, CT45-1, WT-1, FOLR-1, and LAGE-1 [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 C076.