Development of a multi-neoepitope vaccine targeting non-small cell lung cancer through reverse vaccinology and
Elahe Asadollahi1, Alireza Zomorodipour2, Zahra-Soheila Soheili2
1Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Introduction:
Lung cancer, predominantly non-small cell lung cancer (NSCLC), is the leading cause of cancer-related mortality worldwide. Among immunotherapeutic strategies, the personalized multi-neoepitope vaccine (MNEV) offers a promising approach for managing advanced-stage NSCLC.
Methods:
We used reverse vaccinology, immunoinformatics, and bioinformatics to design an MNEV targeting lung cancer in murine (LL/2) cells. Whole exome sequencing (WES) and RNA sequencing data from human and mouse NSCLC cell lines were analyzed to select neoantigens, which were evaluated for their ability to stimulate B cells, helper T lymphocytes (HTLs), and cytotoxic T lymphocytes (CTLs). Molecular docking studies estimated the binding affinity of mouse neoepitopes with MHC class I, MHC class II, and B-cell receptors. Suitable linkers were selected to construct the MNEV, with the 50S L7/L12 ribosomal protein sequence included as an adjuvant to enhance immune responses. The immunoglobulin kappa (Igκ) chain signal peptide was incorporated to improve secretion efficiency. The stability of the final MNEV construct in complex with TLR3, TLR4, and TLR9 was confirmed through binding analysis and refinement of the best-predicted 3D model. To evaluate the immunological efficacy of the MNEV, female C57BL/6 mice were immunized subcutaneously. Immune responses were assessed by measuring total IgG levels in serum using enzyme-linked immunosorbent assay (ELISA) and quantifying IFN-γ and granzyme B levels in the supernatant of cultured splenocytes. The proportions of CD19+ B cells and CD4+ and CD8+ T cells were determined using flow cytometric analysis.
Results:
In silico evaluations indicated that the MNEV is non-toxic, non-allergenic, and stable, exhibiting high-affinity interactions with B lymphocytes, CTLs, and HTLs. Immunization with the MNEV significantly increased serum IgG levels. Flow cytometry analysis revealed higher percentages of CD19+ B cells and CD4+ and CD8+ T cells. Furthermore, splenocytes from immunized mice showed a marked increase in IFN-γ and granzyme B secretion compared to control groups.
Discussion:
This study demonstrates that the MNEV induces a robust strong immune response, highlighting its potential as a promising approach for cancer prevention and immunotherapy, particularly for NSCLC. Furthermore, it provides a foundation for developing neoepitope-based vaccines against various malignancies, guiding future research in cancer vaccine development through advanced computational methods in immunology and oncology.
Insights
This study developed a personalized multi-neoepitope vaccine (MNEV) for lung cancer. The MNEV demonstrated a strong immune response in mice, showing potential for non-small cell lung cancer (NSCLC) immunotherapy.
Area of Science:
- Immunology
- Oncology
- Bioinformatics
Background:
- Non-small cell lung cancer (NSCLC) is a leading cause of cancer mortality globally.
- Personalized multi-neoepitope vaccines (MNEVs) represent a promising immunotherapeutic strategy for advanced NSCLC.
Purpose of the Study:
- To design and evaluate a novel MNEV targeting lung cancer using computational immunology and bioinformatics.
- To assess the MNEV's immunogenicity and potential efficacy in a preclinical murine model.
Main Methods:
- Utilized reverse vaccinology, immunoinformatics, and bioinformatics to select neoantigens from NSCLC cell lines.
- Constructed the MNEV with selected neoantigens, adjuvant (50S L7/L12 ribosomal protein), and signal peptide for enhanced secretion.
- Assessed MNEV stability, binding affinity, and in vivo immunogenicity in C57BL/6 mice through serological, flow cytometry, and cytokine analysis.
Main Results:
- In silico analysis predicted the MNEV to be non-toxic, non-allergenic, and stable with high binding affinity to immune receptors.
- MNEV immunization significantly increased serum IgG levels, B cell (CD19+), helper T cell (CD4+), and cytotoxic T cell (CD8+) proportions.
- Splenocytes from immunized mice exhibited significantly elevated levels of IFN-γ and granzyme B, indicating a robust cellular immune response.
Conclusions:
- The developed MNEV effectively induces a strong and comprehensive immune response in a preclinical setting.
- This study supports the potential of MNEVs as a viable immunotherapy for NSCLC and other malignancies.
- Computational approaches in immunology and oncology are crucial for advancing cancer vaccine development.
