Immunoinformatics-guided design of a multi-epitope vaccine targeting WISP1 for gastric cancer.

Abstract

Background: Gastric cancer remains a leading cause of cancer-related mortality worldwide, and effective preventive or therapeutic vaccines are still lacking. WNT1‑inducible signalling pathway protein 1 (WISP1/CCN4) is a secreted matricellular protein that is overexpressed in gastric tumours and associated with poor prognosis, making it a promising immunotherapy target. We aimed to design a multi‑epitope protein vaccine candidate targeting WISP1 using an immunoinformatics workflow.

Results: We predicted linear B‑cell epitopes from WISP1 and filtered them for antigenicity, non-allergenicity, and non-toxicity. High‑scoring MHC class I and class II T‑cell epitopes with broad HLA coverage were then selected. The final construct combined validated B‑ and T‑cell epitopes with appropriate linkers and a TLR‑agonist adjuvant to enhance immunogenicity. Physicochemical profiling indicated that the construct is stable, soluble, hydrophilic and antigenic, with no predicted allergenicity or toxicity. Secondary and tertiary structures were modelled, refined and validated, revealing proper folding and favourable stereochemical quality. Molecular docking showed strong binding to innate immune receptors, particularly TLR4, and molecular dynamics simulations confirmed stable receptor-vaccine interactions with low structural deviation. Binding‑free energy analysis further supported these results. Immune simulations predicted robust primary and secondary immune responses characterised by sustained IgG/IgM production, increased IFN‑γ and IL‑2, and activation of memory B and T cells. Codon optimisation and in‑silico cloning suggested feasibility for experimental expression.

Conclusions: These results indicate that the proposed WISP1 multi‑epitope vaccine is antigenic, safe, structurally stable and capable of eliciting broad humoral and cellular immune responses in silico. This work provides a testable candidate for experimental validation and highlights secreted CCN family proteins as novel targets for gastric cancer vaccines.