Abstract
The proangiogenic factors including basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) which work synergistically are essential for tumor angiogenesis and tumor growth. The bFGF/VEGF multi-epitope complex peptide (VBP3) was constructed with six different epitope peptides from human bFGF and VEGF. The VBP3 multi-epitope complex peptide was demonstrated good immunogenicity to elicit high titer anti-bFGF antibody and anti-VEGF antibody in C57BL/6 mice. In Lewis lung cancer (LLC) mouse model, the tumor growth was significantly inhibited which resulted in a longer survival time. The immunohistochemistry results showed that the tumor angiogenesis was inhibited with VBP3 vaccine vaccination. The results of flow cytometry (FCM) assay showed that the activation of dentritic cells (DC), CD4+ and CD8+ T cells were stimulated while the infiltration of myeloid-derived suppressor cells and macrophages were suppressed in tumor microenvironment. The polyclonal antibodies were separated from the VBP3-vaccinated mice. The CCK-8 assay results showed that the proliferation of LL-2 cancer cells was inhibited and the Western-blot assay results showed that the phosphorylation levels of Akt and Erk1/2 were decreased by the antibodies. The results indicated that the VBP3 could stimulate specific immune responses to inhibit tumor angiogenesis and tumor growth. The VBP3 with good immunogenicity and immunoprotection could be used as a potential therapeutic peptide vaccine for tumor therapy.
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References
Breslin JW, Pappas PJ, Cerveira JJ, Hobson RW et al (2003) VEGF increases endothelial permeability by separate signaling pathways involving ERK-1/2 and nitric oxide. Am J Physiol Heart Circ Physiol 284:H92-H100. https://doi.org/10.1152/ajpheart.00330.2002
Cao R, Eriksson A, Kubo H, Alitalo K, Cao Y et al (2004) Comparative evaluation of FGF-2-, VEGF-A-, and VEGF-C-induced angiogenesis, lymphangiogenesis,vascular fenestrations, and permeability. Circ Res 94:664–670. https://doi.org/10.1161/01.res.0000118600.91698
Carmeliet P, Jain RK (2011) Molecular mechanisms and clinical applications of angiogenesis. Nature 473:298–307. https://doi.org/10.1038/nature10144
Cheng J, Huo D-M, Kuang D-M, Yang J, Zang L et al (2007) Human macrophages promote the motility and invasiveness of osteopontin-knockdown tumor cells. Cancer Res 67(11):5141–5147. https://doi.org/10.1158/0008-5472.CAN-06-4763
Dailey L, Ambrosetti D, Mansukhani A, Basilico C (2005) Mechanisms underlying differential responses to FGF signaling. Cytokine Growth Factor Rev 16:233–247. https://doi.org/10.1016/j.cytogfr.2005.01.007
Demirkesen C, BüyükpInarbasIlI N, Ramazanoglu R, Oguz O, Mandel NM et al (2006) The correlation of angiogenesis with metastasis in primary cutaneous melanoma: a comparative analysis of microvessel density, expression of vascular endothelial growth factor and basic fibroblastic growth factor. Pathology 38:132–137. https://doi.org/10.1080/00313020600557565
Dikov MM, Ohm JE, Ray N, Tchekneva EE, Burlison J et al (2005) Differential roles of vascular endothelial growth factor receptors 1 and 2 in dendritic cell differentiation. J Immunol 174:215–222. https://doi.org/10.4049/jimmunol.174.1.215
Ferrara N, Gerber H-P, LeCouter J (2003) The biology of VEGF and its receptors. Nat Med 9:669–676. https://doi.org/10.1038/nm0603-669
Finke J, Ko J, Rini B, Rayman P, Ireland J et al (2011) MDSCs as a mechanism of tumor escape from sunitinib mediated anti-angiogenic therapy. Int Immunopharmacol 11:856–861. https://doi.org/10.1016/j.intimp.2011.01.030
Folkman J (2006) Tumor suppression by p53 is mediated in part by the anti-angiogenic activity of endostatin and tumstatin. Science’s STKE. https://doi.org/10.1126/stke.3542006pe35
Foy KC, Miller MJ, Moldovan N, Carson WE, Kaumaya PTP (2012) Combined vaccination with HER-2 peptide followed by therapy with VEGF peptide mimics exerts effective anti-tumor and anti-angiogenic effects in vitro and in vivo. OncoImmunology 1:1048–1060. https://doi.org/10.4161/onci.20708
Gerber H-P, Ferrara N (2005) Pharmacology and pharmacodynamics of bevacizumab as monotherapy or in combination with cytotoxic therapy in preclinical studies. Can Res 65:671–680
Goodman VL, Rock EP, Dagher R, Ramchandani RP, Abraham S et al (2007) Approval summary: sunitinib for the treatment of imatinib refractory or intolerant gastrointestinal stromal tumors and advanced renal cell carcinoma. Clin Cancer Res 13:1367–1373. https://doi.org/10.1634/theoncologist.12-1-107
Grépin R, Pagès G (2010) Molecular mechanisms of resistance to tumour anti-angiogenic strategies. J Oncol. https://doi.org/10.1155/2010/835680
Jekunen A, Kairemo K (2003) Inhibition of angiogenesis at endothelial cell level. Microsc Res Tech 60:85–97. https://doi.org/10.1002/jemt.10247
Kano MR, Morishita Y, Iwata C, Iwasaka S, Watabe T et al (2005) VEGF-A and FGF-2 synergistically promote neoangiogenesis through enhancement of endogenous PDGF-B–PDGFRb signaling. J Cell Sci 118:3759–3768. https://doi.org/10.1242/jcs.02483
Kaumaya PTP, Foy KC (2012) Peptide vaccines and peptidomimetics targeting HER and VEGF proteins may offer a potentially new paradigm in cancer immunotherapy. Future Oncol 8:961–987. https://doi.org/10.2217/fon.12.95
Keedy VL, Sandler AB (2007) Inhibition of angiogenesis in the treatment of non-small cell lung cancer. Cancer Sci 98:1825–1830. https://doi.org/10.1111/j.1349-7006.2007.00620.x
Kerbel RS (2008) Tumor angiogenesis. N Engl J Med 358:2039–2049. https://doi.org/10.1056/NEJMra0706596
Kilvaer TK, Valkov A, Sorbye SW, Smeland E, Bremnes RM, Busund LT, Donnem T (2011) Fibroblast growth factor 2 orchestrates angiogenic networking in non-GIST STS patients. J Transl Med 9:104. https://doi.org/10.1186/1479-5876-9-104
Laxmanan S, Robertson SW, Wang E, Lau JS, Briscoe DM et al (2005) Vascular endothelial growth factor impairs the functional ability of dendritic cells through Id pathways. Biochem Biophys Res Commun 334:193–198. https://doi.org/10.1016/j.bbrc.2005.06.065
Li D, Wang H, Xiang J-J, Deng N, Wang P-P et al (2010) Monoclonal antibodies targeting basic fibroblast growth factor inhibit the growth of B16 melanoma in vivo and in vitro. Oncol Rep 24:457–463. https://doi.org/10.3892/or_00000879
Li Q, Gao S, Yu Y, Wang W, Chen X et al (2012a) A novel bFGF antagonist peptide inhibits breast cancer cell growth. Mol Med Report 6:210–214. https://doi.org/10.3892/mmr.2012.882
Li M, Li M, Zhang H, Luo Z, Wan Y et al (2012b) bFGF peptide combined with the pVAX-8CpG plasmid as adjuvant is a novel anti-cancer vaccine inducing effective immune responses against Lewis lung carcinoma. Mol Med Rep 5:625–630. https://doi.org/10.3892/mmr.2011.725
Lu F, Qin Z-Y, Yang W-B, Qi Y-X, Li Y-M (2004) A DNA vaccine against extracellular domains 1–3 of Flk-1 and its immune preventive and therapeutic effects against H22 tumor cell in vivo. World J Gastroenterol 10:2039–2044. https://doi.org/10.3748/wjg.v10.i14.2039
Ma J, Sawai H, Ochi N, Matsuo Y et al (2009) PTEN regulate angiogenesis through PI3K/Akt/VEGF signaling pathway in human pancreatic cancer cells. Mol Cell Biochem 331:161–171. https://doi.org/10.1007/s11010-009-0154-x
Morera Y, Bequet-Romero M, Ayala M, Velazco JC, Pérez PP et al (2010) Immunogenicity and some safety features of a VEGF-based cancer therapeutic vaccine in rats, rabbits and non-human primates. Vaccine 28:3453–3461. https://doi.org/10.1016/j.vaccine.2010.02.069
Mucha J, Majchrzak K, Taciak B, Hellmén E, Król M (2014) MDSCs mediate angiogenesis and predispose canine mammary tumor cells for metastasis via IL-28/IL-28RA (IFN-l) signaling. PLoS ONE 9(7):e103249. https://doi.org/10.1371/journal.pone.0103249
Nishida T, Ito J, Nagayasu Y, Yokoyama S (2009) FGF-1-induced reactions for biogenesis of apoE-HDL are mediated by Src in rat astrocytes. J Biochem 6:881–886. https://doi.org/10.1093/jb/mvp135
Otrock ZK, Makarem JA, Shamseddine AI (2007) Vascular endothelial growth factor family of ligands and receptors: review. Blood Cells Mol Dis 38:258–268. https://doi.org/10.1016/j.bcmd.2006.12.003
Potente M, Gerhardt H, Carmeliet P (2011) Basic and therapeutic aspects of angiogenesis. Cell 146:873–887. https://doi.org/10.1016/j.cell.2011.08.039
Purcell AW, McCluskey J, Rossjohn J (2007) More than one reason to rethink the use of peptides in vaccine design. Nat Rev Drug Discov 6(5):404–414. https://doi.org/10.1038/nrd2224
Shojaei F, Wu X, Malik AK, Zhong C, Baldwin ME et al (2007) Tumor refractoriness to anti-VEGF treatment is mediated by CD11b+Gr1+myeloid cells. Nat Biotechnol 25:911–920. https://doi.org/10.1038/nbt1323
Suzuki H, Fukuhara M, Yamaura T, Mutoh S, Okabe N et al (2013) 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 11:97. https://doi.org/10.1186/1479-5876-11-97
Tao J, Xiang J-J, Li D, Deng N, Wang H et al (2010) Selection and characterization of a human neutralizing antibody to human fibroblast growth factor-2. Biochem Biophys Res Commun 394:767–773. https://doi.org/10.1016/j.bbrc.2010.03.067
Tartour E, Pere H, Maillere B, Terme M, Merillon N et al (2011) Angiogenesis and immunity: a bidirectional link potentially relevant for the monitoring of anti-angiogenic therapy and the development of novel therapeutic combination with immunotherapy. Cancer Metastasis Rev 30:83–95. https://doi.org/10.1007/s10555-011-9281-4
Terada T, Mizobata M, Kawakami S, Yamashita F, Hashida M (2007) Optimization of tumor-selective targeting by basic fibroblast growth factor-binding peptide grafted PEGylated liposomes. J Controll Rel 119:262–270
Vicari D, Foy KC, Liotta EM, Kaumaya PTP (2011) Engineered conformation-dependent VEGF peptide mimics are effective in inhibiting VEGF signaling pathways. J Biol Chem 286:13612–13625. https://doi.org/10.1074/jbc.M110.216812
Wang H, Zhu Z, Yang Q, Xiang J, Yang H et al (2010) Screening the antigen epitopes of bFGF/VEGF and expressing and identifying their complex peptide. Immunol J 26:688–693
Yamada A, Sasada T, Noguchi M, Itoh K (2013) Next-generation peptide vaccines for advanced cancer. Cancer Sci 104:15–21. https://doi.org/10.1111/cas.12050
Zhang H, Yuan C, Zhang D, Shi H, Li M et al (2011) A novel combined conjugate vaccine: enhanced immunogenicity of bFGF with CRM197 as a carrier protein. Mol Med Rep 4:857–863. https://doi.org/10.3892/mmr.2011.521
Zhou H, Luo Y, Mizutani M, Mizutani N, Dolman C et al (2004) A DNA minigene vaccine against VEGF receptor 2 (Flk-1) suppresses angiogenesis and successfully inhibits growth of prostate and lung carcinoma in mice. Can Res 64:326–326
Acknowledgements
This work was supported by the Grants from State Natural Science Foundation of China (81372281), Science and Technology Planning Project of Guangdong Province (2015B020211009, 2016A010105008) and Science and Technology Planning Project of Guangzhou City (201604020099).
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Ligang Zhang, Dan He, Jianhua Huang, Yanrui Deng, Ruiqiang Weng, Lei Pan, Ning Deng declared that they have no conflict of interest.
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All international, national, and institutional guidelines for the care and use of laboratory animals were followed. This article did not involve any study with human participants.
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Ligang Zhang, Dan He and Jianhua Huang contributed equally to this article.
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Zhang, L., He, D., Huang, J. et al. The Immunogenicity and Immunoprotection of VBP3 Multi-epitope Vaccine Targeting Angiogenesis and Tumor Inhibition in Lung Cancer-Bearing Mice. Int J Pept Res Ther 25, 215–225 (2019). https://doi.org/10.1007/s10989-017-9667-4
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DOI: https://doi.org/10.1007/s10989-017-9667-4