Precision Immunotherapy for NSCLC: Multi-epitope Peptide Vaccine Targeting VEGF-A, TGF-β and MAGE-A3 Increases Antitumor Key Cytokine Balance
Archives of Advances in Biosciences,
Vol. 16 No. 1 (2025),
2 March 2025
,
Page 1-21
https://doi.org/10.22037/aab.v16i1.50519
Abstract
Introduction: Lung cancer remains the leading cause of cancer-related mortality worldwide, underscoring the urgent need for advancements in treatment options. Although, current standard interventions, including surgery, radiotherapy and chemotherapy are widely employed, a significant number of patients experience relapses, highlighting the critical demand for innovative therapeutic strategies. This study was conducted to develop and evaluate a novel multi-epitope peptide vaccine designed from VEGF-A, TGF-β and MAGE-A3 markers, with the aim of enhancing therapeutic efficacy. Lung cancer remains the leading cause of cancer-related mortality worldwide, underscoring the pressing need for more effective therapeutic interventions. Although current standard treatments—including surgery, radiotherapy, and chemotherapy—are widely utilized, a substantial proportion of patients experience disease recurrence, highlighting the necessity for innovative therapeutic strategies. In this study, we developed and evaluated a novel multi-epitope peptide vaccine constructed from VEGF-A, TGF-β, and MAGE-A3 markers, with the objective of enhancing therapeutic efficacy.
Materials and Methods: Optimal epitopes from VEGF-A, TGF-β, and MAGE-A3 were systematically identified and selected, and subsequently conjugated using a KKK linker to form the final multi-epitope vaccine construct. Two groups of BALB/c mice were immunized with the peptide at concentrations of 10 mg/mL and 100 mg/mL, following an immunization protocol that included three weekly administrations. In the fourth week, spleen tissue was collected from the mice to assess the expression levels of IFN-γ, IL-4, IL-6, TNF-α, and IL-10 cytokine genes, thereby enabling a comprehensive evaluation of the immunogenic and functional efficacy of the peptide vaccine.
Results: Bioinformatics evaluations have revealed a promising multi-epitope peptide vaccine,SVRGKGKGQKRKRKKSKKKHHMVKISGGPHISYPPKKKRLESQQTNRRKKRALD.
This peptide notably enhances the expression of key cytokine genes, including TNF-α, IL-6, IFN-γ, IL-4 and IL-10 among the group that received the vaccine at a dose of 10. Even more pronounced levels of gene expression were observed at the higher dose of 100.
Conclusion: This multi-epitope peptide demonstrates considerable potential to elicit a robust immune response and effectively target cancer cells. We strongly recommend conducting further supplementary tests to evaluate its efficacy and possible side effects.
- Multi epitope Peptide Vaccine
- NSCLC
- Cytokine genes
- Immunotherapy
How to Cite
References
1] Herbst RS, Morgensztern D, Boshoff C. The biology and management of non-small cell lung cancer. Nature. 2018 Jan 25;553(7689):446-54. [PMID]
[2] Hu W, Wu H, Li A, Zheng X, Zhang W, Tian Q, Zhang X. Pseudogene CSPG4P12 affects the biological behavior of non small cell lung cancer by Bcl 2/Bax mitochondrial apoptosis pathway. Experimental and Therapeutic Medicine. 2022 Oct 26;24(6):734. [PMID]
[3] Gridelli C, Rossi A, Carbone DP, Guarize J, Karachaliou N, Mok T, Petrella F, Spaggiari L, Rosell R. Non-small-cell lung cancer. Nature reviews Disease primers. 2015 May 21;1(1):1-6.[PMID]
[4] Chen DS, Mellman I. Elements of cancer immunity and the cancer–immune set point. Nature. 2017 Jan 19;541(7637):321-30. [PMID]
[5] Wang XE, Wang YH, Zhou Q, Peng M, Zhang J, Chen M, Ma LJ, Xie GM. Immunomodulatory effect of lentinan on aberrant T subsets and cytokines profile in non-small cell lung cancer patients. Pathology & Oncology Research. 2020 Jan;26(1):499-505. [PMID]
[6] Balkwill F. Tumour necrosis factor and cancer. Nature reviews cancer. 2009 May;9(5):361-71.[PMID]
[7] Benoot T, Piccioni E, De Ridder K, Goyvaerts C. TNFα and immune checkpoint inhibition: friend or foe for lung cancer?. International Journal of Molecular Sciences. 2021 Aug 13;22(16):8691. [PMID]
[8] Ke W, Zhang L, Dai Y. The role of IL‐6 in immunotherapy of non‐small cell lung cancer (NSCLC) with immune‐related adverse events (irAEs). Thoracic cancer. 2020 Apr;11(4):835-9.[PMID]
[9] Guo Y, Xu F, Lu T, Duan Z, Zhang Z. Interleukin-6 signaling pathway in targeted therapy for cancer. Cancer treatment reviews. 2012 Nov 1;38(7):904-10.[PMID]
[10] Fu C, Jiang L, Hao S, Liu Z, Ding S, Zhang W, Yang X, Li S. Activation of the IL-4/STAT6 signaling pathway promotes lung cancer progression by increasing M2 myeloid cells. Frontiers in immunology. 2019 Nov 13;10:2638. [PMID]
[11] Safi S, Yamauchi Y, Hoffmann H, Weichert W, Jost PJ, Winter H, Muley T, Beckhove P. Circulating interleukin-4 is associated with a systemic T cell response against tumor-associated antigens in treatment-naïve patients with resectable non-Small-Cell lung cancer. Cancers. 2020 Nov 24;12(12):3496.[PMID]
[12] Ouyang W, Löhning M, Gao Z, Assenmacher M, Ranganath S, Radbruch A, Murphy KM. Stat6-independent GATA-3 autoactivation directs IL-4-independent Th2 development and commitment. Immunity. 2000 Jan 1;12(1):27-37. [PMID]
[13] Saraiva M, O'garra A. The regulation of IL-10 production by immune cells. Nature reviews immunology. 2010 Mar;10(3):170-81. [PMID]
[14] Couper KN, Blount DG, Riley EM. IL-10: the master regulator of immunity to infection. The Journal of Immunology. 2008 May;180(9):5771-7.[PMID]
[15] Ouyang W, Rutz S, Crellin NK, Valdez PA, Hymowitz SG. Regulation and functions of the IL-10 family of cytokines in inflammation and disease. Annual review of immunology. 2011 Apr 23;29(1):71-109. [PMID]
[16] Yu H, Pardoll D, Jove R. STATs in cancer inflammation and immunity: a leading role for STAT3. Nature reviews cancer. 2009 Nov;9(11):798-809.[PMID]
[17] Hodge G, Barnawi J, Jurisevic C, Moffat D, Holmes M, Reynolds PN, Jersmann H, Hodge S. Lung cancer is associated with decreased expression of perforin, granzyme B and interferon (IFN)-γ by infiltrating lung tissue T cells, natural killer (NK) T-like and NK cells. Clinical & Experimental Immunology. 2014 Oct;178(1):79-85. [PMID]
[18] Jorgovanovic D, Song M, Wang L, Zhang Y. Roles of IFN-γ in tumor progression and regression: a review. Biomarker research. 2020 Sep 29;8(1):49.[PMID]
[19] Wong CW, Huang YY, Hurlstone A. The role of IFN-γ-signalling in response to immune checkpoint blockade therapy. Essays in biochemistry. 2023 Sep;67(6):991-1002. [PMID]
[20] Abd-Aziz N, Poh CL. Development of peptide‐based vaccines for cancer. Journal of Oncology. 2022;2022(1):9749363.[PMID]
[21] Buonaguro L, Tagliamonte M. Peptide-based vaccine for cancer therapies. Frontiers in immunology. 2023 Aug 16;14:1210044. [PMID]
[22] Rahman MM, Masum MH, Talukder A, Akter R. An in silico reverse vaccinology approach to design a novel multiepitope peptide vaccine for non-small cell lung cancers. Informatics in Medicine Unlocked. 2023 Jan 1;37:101169.[LINK]
[23] Peng J, Xiao Y, Wan X, Chen Q, Wang H, Li J, Chen J, Gao R. Enhancement of immune response and anti-infection of mice by porcine antimicrobial peptides and interleukin-4/6 fusion gene encapsulated in chitosan nanoparticles. Vaccines. 2020 Sep 21;8(3):552. [PMID]
[24] Mahaki H, Saeed Modaghegh MH, Nasr Isfahani Z, Amir Daddost R, Molaei P, Ahmadyousefi Y, Vahidzadeh M, Lotfiane E, Tanzadehpanah H. The role of peptide-based tumor vaccines on cytokines of adaptive immunity: a review. International Journal of Peptide Research and Therapeutics. 2021 Dec;27(4):2527-42.[LINK]
[25] Rašková M, Lacina L, Kejík Z, Venhauerova A, Skaličková M, Kolář M, Jakubek M, Rosel D, Smetana Jr K, Brabek J. The role of IL-6 in cancer cell invasiveness and metastasis—overview and therapeutic opportunities. Cells. 2022 Nov 21;11(22):3698. [PMID]
[26] Crotty S. T follicular helper cell differentiation, function, and roles in disease. Immunity. 2014 Oct 16;41(4):529-42.[PMID]
[27] Abbas AK, Benoist C, Bluestone JA, Campbell DJ, Ghosh S, Hori S, Jiang S, Kuchroo VK, Mathis D, Roncarolo MG, Rudensky A. Regulatory T cells: recommendations to simplify the nomenclature. Nature immunology. 2013 Apr;14(4):307-8. [PMID]
[28] Cho HI, Lee YR, Celis E. Interferon γ limits the effectiveness of melanoma peptide vaccines. Blood, The Journal of the American Society of Hematology. 2011 Jan 6;117(1):135-44. [PMID]
[29] Hunter CA, Jones SA. IL-6 as a keystone cytokine in health and disease. Nature immunology. 2015 May;16(5):448-57.[PMID]
[30] Grivennikov SI, Greten FR, Karin M. Immunity, inflammation, and cancer. Cell. 2010 Mar 19;140(6):883-99. [PMID]
[31] Carretero-Iglesia L, Couturaud B, Baumgaertner P, Schmidt J, Maby-El Hajjami H, Speiser DE, Hebeisen M, Rufer N. High peptide dose vaccination promotes the early selection of tumor antigen-specific CD8 T-cells of enhanced functional competence. Frontiers in immunology. 2020 Jan 8;10:3016.[PMID]
[32] Gong W, Liang Y, Mi J, Jia Z, Xue Y, Wang J, Wang L, Zhou Y, Sun S, Wu X. Peptides-based vaccine MP3RT induced protective immunity against Mycobacterium tuberculosis infection in a humanized mouse model. Frontiers in Immunology. 2021 Apr 26;12:666290.[PMID]
[33] Lian F, Yang H, Hong R, Xu H, Yu T, Sun G, Zheng G, Xie B. Evaluation of the antitumor effect of neoantigen peptide vaccines derived from the translatome of lung cancer. Cancer Immunology, Immunotherapy. 2024 May 14;73(7):129. [PMID]
[34] Kumai T, Kobayashi H, Harabuchi Y, Celis E. Peptide vaccines in cancer—old concept revisited. Current opinion in immunology. 2017 Apr 1;45:1-7.[PMID]
[35] Yang L, Wang L, Zhang Y. Immunotherapy for lung cancer: advances and prospects. American journal of clinical and experimental immunology. 2016 Mar 23;5(1):1. [PMID]
[36] Hemmati S, Saeidikia Z, Seradj H, Mohagheghzadeh A. Immunomodulatory peptides as vaccine adjuvants and antimicrobial agents. Pharmaceuticals. 2024 Feb 2;17(2):201. [PMID]
[37] Eisenbarth SC, Colegio OR, O’Connor W, Sutterwala FS, Flavell RA. Crucial role for the Nalp3 inflammasome in the immunostimulatory properties of aluminium adjuvants. Nature. 2008 Jun 19;453(7198):1122-6.[LINK]
[38] Kool M, Soullié T, Van Nimwegen M, Willart MA, Muskens F, Jung S, Hoogsteden HC, Hammad H, Lambrecht BN. Alum adjuvant boosts adaptive immunity by inducing uric acid and activating inflammatory dendritic cells. The Journal of experimental medicine. 2008 Apr 14;205(4):869-82. [PMID]
[39] HogenEsch H. Mechanism of immunopotentiation and safety of aluminum adjuvants. Frontiers in immunology. 2013 Jan 10;3:406.[PMID]
[40] Moyer TJ, Zmolek AC, Irvine DJ. Beyond antigens and adjuvants: formulating future vaccines. The Journal of clinical investigation. 2016 Mar 1;126(3):799-808.[PMID]
[41] Heidary F, Tourani M, Hejazi-Amiri F, Khatami SH, Jamali N, Taheri-Anganeh M. Design of a new multi-epitope peptide vaccine for non-small cell Lung cancer via vaccinology methods: an in silico study. Molecular Biology Research Communications. 2022 Mar;11(1):55.[PMID]
[42] Van Horssen R, Ten Hagen TL, Eggermont AM. TNF-α in cancer treatment: molecular insights, antitumor effects, and clinical utility. The oncologist. 2006 Apr 1;11(4):397-408. [PMID]
[43] Lawrence T. The nuclear factor NF-κB pathway in inflammation. Cold Spring Harbor perspectives in biology. 2009 Dec 1;1(6):a001651.[PMID]
[44] Tanaka T, Narazaki M, Kishimoto T. IL-6 in inflammation, immunity, and disease. Cold Spring Harbor perspectives in biology. 2014 Oct 1;6(10):a016295. [PMID]
[45] Karakasheva TA, Lin EW, Tang Q, Qiao E, Waldron TJ, Soni M, Klein-Szanto AJ, Sahu V, Basu D, Ohashi S, Baba K. IL-6 mediates cross-talk between tumor cells and activated fibroblasts in the tumor microenvironment. Cancer Research. 2018 Sep 1;78(17):4957-70.[PMID]
[46] Ni L, Lu J. Interferon gamma in cancer immunotherapy. Cancer medicine. 2018 Sep;7(9):4509-16.[PMID]
[47] Castro F, Cardoso AP, Gonçalves RM, Serre K, Oliveira MJ. Interferon-gamma at the crossroads of tumor immune surveillance or evasion. Frontiers in immunology. 2018 May 4;9:847.[PMID]
[48] Zhang Y, Li D, Shen Y, Li S, Lu S, Zheng B. Immunization with a novel mRNA vaccine, TGGT1_216200 mRNA-LNP, prolongs survival time in BALB/c mice against acute toxoplasmosis. Frontiers in immunology. 2023 Apr 14;14:1161507.[PMID]
[49] Moore KW, de Waal Malefyt R, Coffman RL, O'Garra A. Interleukin-10 and the interleukin-10 receptor. Annual review of immunology. 2001 Apr;19(1):683-765.[PMID]
[50] Dent AL, Hu-Li J, Paul WE, Staudt LM. T helper type 2 inflammatory disease in the absence of interleukin 4 and transcription factor STAT6. Proceedings of the National Academy of Sciences. 1998 Nov 10;95(23):13823-8.[PMID]
[51] Eisenbarth SC. Dendritic cell subsets in T cell programming: location dictates function. Nature Reviews Immunology. 2019 Feb;19(2):89-103.[PMID]
[52] Heinrich PC, Behrmann I, Müller-Newen G, Schaper F, Graeve L. Interleukin-6-type cytokine signalling through the gp130/Jak/STAT pathway. Biochemical journal. 1998 Sep 1;334(2):297-314.[PMID]
[53] Murray PJ. Understanding and exploiting the endogenous interleukin-10/STAT3-mediated anti-inflammatory response. Current opinion in pharmacology. 2006 Aug 1;6(4):379-86. [PMID]
[54] Hu X, Li J, Fu M, Zhao X, Wang W. The JAK/STAT signaling pathway: from bench to clinic. Signal transduction and targeted therapy. 2021 Nov 26;6(1):402.[PMID]
[55] Bergmann-Leitner ES, Chaudhury S, Steers NJ, Sabato M, Delvecchio V, Wallqvist AS, Ockenhouse CF, Angov E. Computational and experimental validation of B and T-cell epitopes of the in vivo immune response to a novel malarial antigen. PloS one. 2013 Aug 16;8(8):e71610.[PMID]
[56] Khanum S, Carbone V, Gupta SK, Yeung J, Shu D, Wilson T, Parlane NA, Altermann E, Estein SM, Janssen PH, Wedlock DN. Mapping immunogenic epitopes of an adhesin-like protein from Methanobrevibacter ruminantium M1 and comparison of empirical data with in silico prediction methods. Scientific Reports. 2022 Jun 21;12(1):10394.[PMID]
[57] Wang T, Chen S, Wang Y, Zhang Y, Song X, Bi Z, Liu M, Niu Q, Liu J, Feng P, Sun X. From In Silico to In Vitro: A Comprehensive Guide to Validating Bioinformatics Findings. arXiv preprint arXiv:2502.03478. 2025 Jan 24. [LINK]
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