Induction of humoral immune responses and inhibition of metastasis in mice by a VEGF peptide-based vaccine

Document Type : Original Article

Authors

1 Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Immunology Research Center, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

3 Department of Pathology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

4 Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Objective(s): Blocking of vascular endothelial growth factor (VEGF) plays a pivotal role in inhibition of metastasis and is a target for development of anti-angiogenic agents. In this study, a peptide-based vaccine was designed and its potential for induction of humoral immune responses, generation of neutralizing antibodies, inhibition of tumor growth and metastasis was determined.
Materials and Methods: With online bioinformatics tools, a fragment of the VEGF-A was selected for a peptide-based vaccine. To enhance its antigenicity, the peptide was conjugated with Keyhole limpet hemocyanin and used to immunize mice. Then, the polyclonal anti-VEGF antibody titer was measured and its effect on proliferation of HUVEC cell line was investigated by MTT assay. Finally, we checked the effect of the peptide on tumor growth, metastasis, and survival rates in a mouse model of cancer.
Results: The bioinformatics analysis of the selected region confirmed dis-similarity of the peptide with any other human protein and its acceptable antigenicity to stimulate a tumor-specific humoral response. Anti-VEGF antibody titers were significantly greater in vaccinated mice than in controls. IgG antibody from mice immunized with recombinant VEGF-A inhibited HUVEC proliferation (P<0.0001). Tumors in vaccinated mice were significantly smaller than those in controls. Moreover, metastasis was reduced and survival rates increased in the vaccinated group.
Conclusion: Production of high-titer antibody against the peptide vaccine indicated that the peptide has the potency to be used as a peptide-based vaccine for humoral inhibition of tumor growth and metastasis. The efficacy of the peptide should be further tested in primate models.

Keywords

References

1. Folkman J. Tumor angiogenesis: therapeutic implications. N Engl J Med 1971;285:1182-1186.
2. Ramjiawan RR, Griffioen AW, Duda DG. Anti-angiogenesis for cancer revisited: Is there a role for combinations with immunotherapy? Angiogenesis 2017;20:185-204.
3. Tugues S, Koch S, Gualandi L, Li X, Claesson-Welsh L. Vascular endothelial growth factors and receptors: anti-angiogenic therapy in the treatment of cancer. Mol Aspects Med 2011;32:88-111.
4. Carmeliet P, Jain RK. Molecular mechanisms and clinical applications of angiogenesis. Nature 2011;473:298-307.
5. Woolard J, Bevan HS, Harper SJ, Bates DO. Molecular diversity of VEGF-A as a regulator of its biological activity. Microcirculation 2009;16:572-592.
6. Ellis LM, Hicklin DJ. VEGF-targeted therapy: mechanisms of anti-tumour activity. Nature Rew Cancer 2008;8:579-591.
7. Heath VL, Bicknell R. Anticancer strategies involving the vasculature. Nature Rew Clin Oncol 2009;6:395-404.
8. Kong DH, Kim MR, Jang JH, Na HJ, Lee S. A review of anti-angiogenic targets for monoclonal antibody cancer therapy. Int J Mol Sci 2017;18. pii: E1786.
9. Matrana MR, Atkinson B, Jonasch E, Tannir NM. Emerging targeted therapies in metastatic renal cell carcinoma. Curr Clin Pharmacol 2011;6:189-198.
10. Hurwitz H, Fehrenbacher L, Novotny W, Cartwright T, Hainsworth J, Heim W, et al. Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N Engl J Med 2004;350:2335-2342.
11. Emens LA. Cancer vaccines: on the threshold of success. Expert Opin Emerg Drugs 2008;13:295-308.
12. Berzofsky JA, Oh S, Terabe M. Peptide vaccines against cancer. Cancer Treat Res 2005;123:115-136.
13. Morera Y, Bequet-Romero M, Ayala M, Lamdan H, Agger EM, Andersen P, et al. Anti-tumoral effect of active immunotherapy in C57BL/6 mice using a recombinant human VEGF protein as antigen and three chemically unrelated adjuvants. Angiogenesis 2008;11:381-393.
14. Morera Y, Bequet-Romero M, Ayala M, Perez PP, Castro J, Sanchez J, et al. Antigen dose escalation study of a VEGF-based therapeutic cancer vaccine in non human primates. Vaccine 2012;30:368-377.
15. Okaji Y, Tsuno NH, Saito S, Yoneyama S, Tanaka M, Nagawa H, et al. Vaccines targeting tumour angiogenesis: a novel strategy for cancer immunotherapy. European J Surgic Oncol 2006;32:363-370.
16. Gavilondo JV, Hernandez-Bernal F, Ayala-Avila M, de la Torre AV, de la Torre J, Morera-Diaz Y, et al. Specific active immunotherapy with a VEGF vaccine in patients with advanced solid tumors. results of the CENTAURO antigen dose escalation phase I clinical trial. Vaccine 2014;32:2241-2250.
17. Soltanpour-Gharibdousti F, Kardar GA, Fazeli-Delshad B, Falak R, Ganjalikhani-Hakemi M, Andalib A. Bioinformatic designing for producing vaccine peptide of human vascular endothelial growth factor (VEGF-A), and evaluation of polyclonal antibodies in mice. J Isfahan Med Sch 2016;34:1054-1059.
18. Soukhtanloo M, Falak R, Sankian M, Varasteh AR. Generation and characterization of anti-chitinase monoclonal antibodies. Hybridoma 2011;30:145-151.
19. Fazeli-Delshad B, Ganjalikhani-Hakemi M, Soltanpour-Gharibdousti F, Shayanfar N, Falak N, Kardar GA. Evaluation of the functionality of 4T1 cell line in development of metastatic cancer in BALB/c mice as an animal model. Journal of Isfahan Medical School 2016;34:985-990.
20. Briand JP, Muller S, Van Regenmortel MHV. Synthetic peptides as antigens: Pitfalls of conjugation methods. J Immunol Methods 1985;78:59-69.
21. Zegers N, Gerritse K, Deen C, Boersma W, Claassen E. An improved conjugation method for controlled covalent coupling of synthetic peptides to proteins using glutaraldehyde in a dialysis method. J Immunol Methods 1990;130:195-200.
22. Swain M, Ross NW. A silver stain protocol for proteins yielding high resolution and transparent background in sodium dodecyl sulfate-polyacrylamide gels. Electrophoresis 1995;16:948-951.
23. Ichihara E, Kiura K, Tanimoto M. Targeting Angiogenesis in Cancer Therapy. Acta Med Okayama 2011;65:353-62.
24. Bellou S, Pentheroudakis G, Murphy C, Fotsis T. Anti-angiogenesis in cancer therapy: Hercules and hydra. Cancer lett 2013;338:219-228.
25. Suzuki H, Fukuhara M, Yamaura T, Mutoh S, Okabe N, Yaginuma H, et al. Multiple therapeutic peptide vaccines consisting of combined novel cancer testis antigens and anti-angiogenic peptides for patients with non-small cell lung cancer. J Transl Med 2013;11:97.
26. Vacchelli E, Martins I, Eggermont A, Fridman W, Galon J, Sautès-Fridman C, et al. Trial watch: Peptide vaccines in cancer therapy. Oncoimmunology 2012;1:1557-1576.
27. Wei YQ, Huang MJ, Yang L, Zhao X, Tian L, Lu Y, et al. Immunogene therapy of tumors with vaccine based on Xenopus homologous vascular endothelial growth factor as a model antigen. Proc Natl Acad Sci USA 2001;98:11545-11550.
28. Kamstock D, Elmslie R, Thamm D, Dow S. Evaluation of a xenogeneic VEGF vaccine in dogs with soft tissue sarcoma. Cancer Immunol Immunother 2007;56:1299-1309.
29. Rad FH, Le Buanec H, Paturance S, Larcier P, Genne P, Ryffel B, et al. VEGF kinoid vaccine, a therapeutic approach against tumor angiogenesis and metastases. Proc Natl Acad Sci USA 2007;104:2837-2842.
30. Morera Y, Bequet-Romero M, Ayala M, Velazco JC, Perez PP, Alba JS, et al. Immunogenicity and some safety features of a VEGF-based cancer therapeutic vaccine in rats, rabbits and non-human primates. Vaccine 2010;28:3453-3461.
31. Morera Y, Gonzalez R, Lamdan H, Perez L, Gonzalez Y, Aguero J, et al. Vaccination with a mutated variant of human Vascular Endothelial Growth Factor (VEGF) blocks VEGF-induced retinal neovascularization in a rabbit experimental model. Exp Eye Res 2014;122:102-109.
32. Vicari D, Foy KC, Liotta EM, Kaumaya PT. Engineered conformation-dependent VEGF peptide mimics are effective in inhibiting VEGF signaling pathways. J Biol Chem 2011;286:13612-13625.
33. Wang B, Kaumaya PT, Cohn DE. Immunization with synthetic VEGF peptides in ovarian cancer. Gynecol Oncol 2010;119:564-570.
34. Kyutoku M, Nakagami H, Koriyama H, Tomioka H, Nakagami F, Shimamura M, et al. Development of novel DNA vaccine for VEGF in murine cancer model. Sci Rep 2013;3:3380.
35. Pulaski BA, Ostrand-Rosenberg S. Reduction of established spontaneous mammary carcinoma metastases following immunotherapy with major histocompatibility complex class II and B7.1 cell-based tumor vaccines. Cancer Res 1998;58:1486-1493.
36. Lelekakis M, Moseley JM, Martin TJ, Hards D, Williams E, Ho P, et al. A novel orthotopic model of breast cancer metastasis to bone. Clin Exp Metastasis 1999;17:163-170.
37. Mawalla B, Yuan X, Luo X, Chalya PL. Treatment outcome of anti-angiogenesis through VEGF-pathway in the management of gastric cancer: a systematic review of phase II and III clinical trials. BMC Res Notes 2018;11:21.
38. Wada S, Yada E, Ohtake J, Sasada T. Personalized peptide vaccines for cancer therapy: current progress and state of the art. Expert Rev Precis Med Drug Dev 2017;2:371-381.
39. Vreeland TJ, Hale DK, Clifton GT, Sears AK, Mittendorf EA and Peoples GE. Peptide vaccine strategies in the treatment of cancer. J Proteomics Bioinform 2013;6:081-084.
Iranian Journal of Basic Medical Sciences
Volume 23, Issue 4
April 2020
Pages 507-514

Files

Share

How to cite

Statistics