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Vaccination with a potent DNA vaccine targeting B-cell epitopes of hGRP induces prophylactic and therapeutic antitumor activity in vivo

Gene Therapy volume 17, pages 459–468 (2010)Cite this article

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Abstract

Gastrin-releasing peptide (GRP), a bombesin-like peptide, is an autocrine or paracrine growth factor that can stimulate the growth of various cancer cells, making it an ideal target antigen to develop vaccines against cancer. In this study, we developed a novel DNA vaccine that encodes six tandem repeats of B-cell epitope GRP18−27 (GRP6) flanked by HSP65 as carrier and four tandem repeats of mycobacterial HSP70407−426 (M4) as helper T-cell epitopes for enhancement of immunogenicity. When intramuscularly immunized to mice, this anti-GRP DNA vaccine-induced GRP-specific antibody (Ab) responses that were at least 10-fold higher in magnitude compared with HSP65–GRP6 protein vaccine. Both prophylactic and therapeutic antitumor immunities induced by vaccination significantly suppressed the growth of GRP-dependent prostate carcinoma RM-1 in vivo and prolonged the survival of tumor-inoculated mice. Out results also showed that the immune sera with high titer of GRP-specific Abs effectively inhibited the growth of tumor in mice and dose dependently inhibited proliferation of cultured RM-1 cells in vitro, suggesting that the GRP neutralizing Ab is responsible for the protective and therapeutic antitumor activity of vaccination. These findings may be of great importance in the further exploration of the applications of growth factors identified in human in cancer therapy.

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References

  1. Patel O, Shulkes A, Baldwin GS . Gastrin-releasing peptide and cancer. Biochim Biophys Acta 2006; 1766: 23–41.

    CAS  PubMed  Google Scholar 

  2. Xiao D, Qu X, Weber HC . GRP receptor-mediated immediate early gene expression and transcription factor Elk-1 activation in prostate cancer cells. Regul Pept 2002; 109: 141–148.

    Article  CAS  Google Scholar 

  3. Hellmich MR, Ives KL, Udupi V, Soloff MS, Greeley Jr GH, Christensen BN et al. Multiple protein kinase pathways are involved in gastrin-releasing peptide receptor-regulated secretion. J Biol Chem 1999; 274: 23901–23909.

    Article  CAS  Google Scholar 

  4. Aprikian AG, Tremblay L, Han K, Chevalier S . Bombesin stimulates the motility of human prostate-carcinoma cells through tyrosine phosphorylation of focal adhesion kinase and of integrin-associated proteins. Int J Cancer 1997; 72: 498–504.

    Article  CAS  Google Scholar 

  5. Miyazaki M, Lamharzi N, Schally AV, Halmos G, Szepeshazi K, Groot K et al. Inhibition of growth of MDA-MB-231 human breast cancer xenografts in nude mice by bombesin/gastrin-releasing peptide (GRP) antagonists RC-3940-II and RC-3095. Eur J Cancer 1998; 34: 710–717.

    Article  CAS  Google Scholar 

  6. Moody TW, Jensen RT, Garcia L, Leyton J . Nonpeptide neuromedin B receptor antagonists inhibit the proliferation of C6 cells. Eur J Pharmacol 2000; 409: 133–142.

    Article  CAS  Google Scholar 

  7. Moody TW, Leyton J, Garcia-Marin L, Jensen RT . Nonpeptide gastrin releasing peptide receptor antagonists inhibit the proliferation of lung cancer cells. Eur J Pharmacol 2003; 474: 21–29.

    Article  CAS  Google Scholar 

  8. Chaudhry A, Carrasquillo JA, Avis IL, Shuke N, Reynolds JC, Bartholomew R et al. Phase I and imaging trial of a monoclonal antibody directed against gastrin-releasing peptide in patients with lung cancer. Clin Cancer Res 1999; 5: 3385–3393.

    CAS  PubMed  Google Scholar 

  9. Szepeshazi K, Schally AV, Nagy A, Wagner BW, Bajo AM, Halmos G . Preclinical evaluation of therapeutic effects of targeted cytotoxic analogs of somatostatin and bombesin on human gastric carcinomas. Cancer 2003; 98: 1401–1410.

    Article  CAS  Google Scholar 

  10. Moody TW, Mantey SA, Pradhan TK, Schumann M, Nakagawa T, Martinez A et al. Development of high affinity camptothecin-bombesin conjugates that have targeted cytotoxicity for bombesin receptor-containing tumor cells. J Biol Chem 2004; 279: 23580–23589.

    Article  CAS  Google Scholar 

  11. Gonzalez G, Lage A . Cancer vaccines for hormone/growth factor immune deprivation: a feasible approach for cancer treatment. Curr Cancer Drug Targets 2007; 7: 229–241.

    Article  CAS  Google Scholar 

  12. Guojun W, Wei G, Kedong O, Yi H, Yanfei X, Qingmei C et al. A novel vaccine targeting gastrin-releasing peptide: efficient inhibition of breast cancer growth in vivo. Endocr Relat Cancer 2008; 15: 149–159.

    Article  Google Scholar 

  13. Anand SB, Murugan V, Prabhu PR, Anandharaman V, Reddy MV, Kaliraj P . Comparison of immunogenicity, protective efficacy of single and cocktail DNA vaccine of Brugia malayi abundant larval transcript (ALT-2) and thioredoxin peroxidase (TPX) in mice. Acta Trop 2008; 107: 106–112.

    Article  CAS  Google Scholar 

  14. Nimal S, Heath AW, Thomas MS . Enhancement of immune responses to an HIV gp120 DNA vaccine by fusion to TNF alpha cDNA. Vaccine 2006; 24: 3298–3308.

    Article  CAS  Google Scholar 

  15. Lu Y, Ouyang K, Fang J, Zhang H, Wu G, Ma Y et al. Improved efficacy of DNA vaccination against prostate carcinoma by boosting with recombinant protein vaccine and by introduction of a novel adjuvant epitope. Vaccine 2009; 27: 5411–5418.

    Article  CAS  Google Scholar 

  16. Papac DI, Hoyes J, Tomer KB . Epitope mapping of the gastrin-releasing peptide/anti-bombesin monoclonal antibody complex by proteolysis followed by matrix-assisted laser desorption ionization mass spectrometry. Protein Sci 1994; 3: 1485–1492.

    Article  CAS  Google Scholar 

  17. Perraut R, Lussow AR, Gavoille S, Garraud O, Matile H, Tougne C et al. Successful primate immunization with peptides conjugated to purified protein derivative or mycobacterial heat shock proteins in the absence of adjuvants. Clin Exp Immunol 1993; 93: 382–386.

    Article  CAS  Google Scholar 

  18. Wang Y, Whittall T, McGowan E, Younson J, Kelly C, Bergmeier LA et al. Identification of stimulating and inhibitory epitopes within the heat shock protein 70 molecule that modulate cytokine production and maturation of dendritic cells. J Immunol 2005; 174: 3306–3316.

    Article  CAS  Google Scholar 

  19. Jin L, Wang Y, Xiong Q, Chen Q, Li J, Zhu A et al. Long-lasting specific antibodies against P277 induced by mucosal administration of P277 repeat sequences carried by Hsp65 in the absence of adjuvants. Vaccine 2007; 25: 2043–2050.

    Article  CAS  Google Scholar 

  20. Dupuis M, Denis-Mize K, Woo C, Goldbeck C, Selby MJ, Chen M et al. Distribution of DNA vaccines determines their immunogenicity after intramuscular injection in mice. J Immunol 2000; 165: 2850–2858.

    Article  CAS  Google Scholar 

  21. Kjerrulf M, Lowenadler B, Svanholm C, Lycke N . Tandem repeats of T helper epitopes enhance immunogenicity of fusion proteins by promoting processing and presentation. Mol Immunol 1997; 34: 599–608.

    Article  CAS  Google Scholar 

  22. Yankai Z, Rong Y, Yi H, Wentao L, Rongyue C, Ming Y et al. Ten tandem repeats of beta-hCG 109–118 enhance immunogenicity and anti-tumor effects of beta-hCG C-terminal peptide carried by mycobacterial heat-shock protein HSP65. Biochem Biophys Res Commun 2006; 345: 1365–1371.

    Article  Google Scholar 

  23. Pullen GR, Fitzgerald MG, Hosking CS . Antibody avidity determination by ELISA using thiocyanate elution. J Immunol Methods 1986; 86: 83–87.

    Article  CAS  Google Scholar 

  24. Jin L, Zhu A, Wang Y, Chen Q, Xiong Q, Li J et al. A Th1-recognized peptide P277, when tandemly repeated, enhances a Th2 immune response toward effective vaccines against autoimmune diabetes in nonobese diabetic mice. J Immunol 2008; 180: 58–63.

    Article  CAS  Google Scholar 

  25. Kroog GS, Sainz E, Worland PJ, Akeson MA, Benya RV, Jensen RT et al. The gastrin-releasing peptide receptor is rapidly phosphorylated by a kinase other than protein kinase C after exposure to agonist. J Biol Chem 1995; 270: 8217–8224.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the China National Natural Science Fund Committee (grant nos. 30500458, 30701023, 30672464, 30572272, 30872393 and 30772570); the Natural Science Foundation of Jiangsu Province (no. BK 2007170 and BK2007169).

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Authors and Affiliations

  1. Minigene Pharmacy Laboratory, Biopharmaceutical College, China Pharmaceutical University, Nanjing, China

    Lu Yong, Zhang Huiyong, Hou Jing, Wang Huaqian, Hu Xiangbing, Ma Yanjun, Ge Xiaoyu, Huang Li, Yang Yanan, Cao Rongyue, Liu Jingjing & Wu Jie

  2. Department of Life Sciences and Technology, Xinxiang Medical University, Xinxiang, China

    Zhang Huiyong

  3. Shanghai Yijiu Biomedical Cooperation Ltd, Shanghai Zhangjiang Hi-Tech Park, Shanghai, China

    Lu Yong & Fan Hao

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  1. Lu Yong
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  2. Zhang Huiyong
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Correspondence to Wu Jie.

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Yong, L., Huiyong, Z., Jing, H. et al. Vaccination with a potent DNA vaccine targeting B-cell epitopes of hGRP induces prophylactic and therapeutic antitumor activity in vivo. Gene Ther 17, 459–468 (2010). https://doi.org/10.1038/gt.2009.165

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  • DOI: https://doi.org/10.1038/gt.2009.165

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