Construction of a Recombinant Phage-vaccine Capable of Reducing the Growth Rate of an Established LL2 Tumor Model
Over expression of the epidermal growth factor receptor (EGFR) in many human epithelial tumors has been correlated with disease progression and poor prognosis. EGFR-inhibiting immunotherapy has already been introduced in cancer therapy. Peptide displaying phage particles in eukaryotic hosts can behave as antigen carriers, able to activate the innate immune system and to elicit adaptive immunity. Herein, the M13-pAK8-VIII phagemid plasmid was engineered to contain the sequences for an EGFR mimotope along with the L2 extracellular domain of EGFR (EM-L2) which would produce the final peptide-phage vaccine. The prophylactic and therapeutic effects of this novel vaccine were evaluated on the Lewis lung carcinoma induced mouse (C57/BL6) model. The recombinant peptide was confirmed to be displayed on the surface of M13 phage as an extension for phage’s PVIII protein. Immunization of mice with peptide-phage vaccine resulted in antibody production against EM-L2 and significant reduction of tumor growth rate by nearly 25 percent. In conclusion, EM-L2 displaying phage particles could be deemed as an encouraging strategy in contemporary cancer immunotherapy.
1. Troiani T, Zappavigna S, Martinelli E, Addeo SR, Stiuso P, Ciardiello F, et al. Optimizing treatment of metastatic colorectal cancer patients with anti-EGFR antibodies: overcoming the mechanisms of cancer cell resistance. Expert Opin Biol Ther 2013; 13(2):241-55.
2. Najar AG, Pashaei-Asl R, Omidi Y, Farajnia S, Nourazarian AR. EGFR antisense oligonucleotides encapsulated with nanoparticles decrease EGFR, MAPK1 and STAT5 expression in a human colon cancer cell line. Asian Pac J Cancer Prev 2013; 14(1):495-8.
3. Jorissen RN, Treutlein HR, Burgess AW, Epa VC, Garrett TP, Ward CW. Characterization of a comparative model of the extracellular domain of the epidermal growth factor receptor. Protein Sci 2000; 9(2):310-24.
4. Lax I, Fischer R, Ng C, Segre J, Ullrich A, Givol D, et al. Noncontiguous regions in the extracellular domain of EGF receptor define ligand-binding specificity. Cell Regul 1991; 2(5):337-45.
5. Lemmon MA, Bu Z, Ladbury JE, Zhou M, Pinchasi D, Lax I, et al. Two EGF molecules contribute additively to stabilization of the EGFR dimer. EMBO J 1997; 16(2):281-94.
6. Ramírez BS, Pestana ES, Hidalgo GG, García TH, Rodríguez RP, Ullrich A, et al. Active antimetastatic immunotherapy in Lewis lung carcinoma with self EGFR extracellular domain protein in VSSP adjuvant. Int J Cancer 2006; 119(2):2190-9.
7. Woodburn J. The epidermal growth factor receptor and its inhibition in cancer therapy. Pharmacol Ther 1999; 82(2-3):241-50.
8. Laird AD, Cherrington JM. Small molecule tyrosine kinase inhibitors: clinical development of anticancer agents. Expert Opin Investig Drugs 2003; 12(1):51-64.
9. B Yousefi H, Yuan J, Keshavarz-Fathi M, Murphy JF, Rezaei N. Immunotherapy of cancers comes of age. Expert Rev Clin Immunol 2017; 13(10):1001-15.
10. Asadi-Ghalehni M, Rasaee MJ, RajabiBazl M, et al. A novel recombinant anti-epidermal growth factor receptor peptide vaccine capable of active immunization and reduction of tumor volume in a mouse model. Microbiol Immunol 2017; 61(12):531-8.
11. Smith GP. Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 1985; 228(4705):1315-8.
12. Navari M, Zare M, Javanmardi M, Asadi-Ghalehni M, Modjtahedi H, Rasaee MJ. Epitope mapping of epidermal growth factor receptor (EGFR) monoclonal antibody and induction of growth-inhibitory polyclonal antibodies by vaccination with EGFR mimotope. Immunopharmacol Immunotoxicol 2014; 36(5):309-15.
13. Asadi-Ghalehni M, Ghaemmaghami M, Klimka A, Javanmardi M, Navari M, Rasaee MJ. Cancer immunotherapy by a recombinant phage vaccine displaying EGFR mimotope: an in vivo study. Immunopharmacol Immunotoxicol 2015; 37(3):274-9.
14. Javanmardi M, Modjtahedi H, Asadi-Ghalehni M, Maghami MG. Triple tandem mimotope peptide of Epidermal Growth Factor Receptor displaying on the surface of M13 phage induces anti-tumor response in mice tumor model. Iranian Journal of Biotechnology 2014; 12:9-17.
15. Aghebati-Maleki L, Bakhshinejad B, Baradaran B, et al. Phage display as a promising approach for vaccine development. J Biomed Sci 2016; 23(1):66.
16. Sharav T, Wiesmüller K-H, Walden P. Mimotope vaccines for cancer immunotherapy. Vaccine 2007; 25:3032-7.
17. Campos‐Perez J, Rice J, Escors D, Collins M, Paterson A, Savelyeva N, et al. DNA fusion vaccine designs to induce tumor‐lytic CD8+ T‐cell attack via the immunodominant cysteine‐containing epitope of NY‐ESO 1. Int J Cancer 2013; 133(6):1400-7.
18. Javanmardi M, Rasaee mj, Modjtahedi H, Asadi-Ghalehni M, Maghami MG. Triple tandem mimotope peptide of Epidermal Growth Factor Receptor displaying on the surface of M13 phage induces anti-tumor response in mice tumor model. Iranian Journal of Biotechnology 2014; 12:9-17.
19. Wu Y, Wan Y, Bian J, Zhao J, Jia Z, Zhou L, et al. Phage display particles expressing tumor‐specific antigens induce preventive and therapeutic anti‐tumor immunity in murine p815 model. Int J Cancer 2002; 98(5):748-53.
20. Asadi-Ghalehni M, Rasaee MJ, Javanmardi M, Khalili S, Mohamadi M, Fatemi F. in Silico and in Vitro Evaluation of A Recombinant Fusion Peptide as A Novel Candidate Vaccine for EGFR-Positive Tumors. Biosciences Biotechnology Research Asia 2015; 12:2405-10.
21. Duś D, Budzyński W, Radzikowski C. LL2 cell line derived from transplantable murine Lewis lung carcinoma--maintenance in vitro and growth characteristics. Arch Immunol Ther Exp (Warsz) 1984; 33(6):817-23.
22. Khalili S, Jahangiri A, Hashemi ZS, Khalesi B, Mardsoltani M, Amani J. Structural pierce into molecular mechanism underlying Clostridium perfringens Epsilon toxin function. Toxicon 2017; 127:90-99.
23. Khalili S, Rasaee M, Bamdad T. 3D structure of DKK1 indicates its involvement in both canonical and non-canonical Wnt pathways. Mol Biol (Mosk) 2017; 51(1):155-66.
24. Jahangiri A, Rasooli I, Owlia P, Fooladi AAI, Salimian J. In silico design of an immunogen against Acinetobacter baumannii based on a novel model for native structure of outer membrane protein A. Microb Pathog 2017; 105:201-10.
25. Khalili S, Mohammadpour H, Shokrollahi Barough M, Kokhaei P. ILP-2 modeling and virtual screening of an FDA-approved library: a possible anticancer therapy. Turk J Med Sci 2016; 46(4):1135-43.
26. Mohammadpour H, Pourfathollah AA, Zarif MN, Khalili S. Key role of Dkk3 protein in inhibition of cancer cell proliferation: an in silico identification. J Theor Biol 2016; 393:98-104.
27. Mohammadpour H, Khalili S, Hashemi ZS. Kremen is beyond a subsidiary co-receptor of Wnt signaling: an in silico validation. Turkish Journal of Biology 2015; 39:501-10.
28. Khalili S, Zakeri A, Hashemi ZS, Masoumikarimi M, Manesh MRR, Shariatifar N, et al. Structural analyses of the interactions between the thyme active ingredients and human serum albumin. Turkish Journal of Biochemistry 2017; 42:459-67.
29. Khalili S, Rahbar MR, Dezfulian MH, Jahangiri A. In silico analyses of Wilms׳ tumor protein to designing a novel multi-epitope DNA vaccine against cancer. J Theor Biol 2015; 379:66-78.
30. Khalili S, Jahangiri A, Borna H, Ahmadi Zanoos K, Amani J. Computational vaccinology and epitope vaccine design by immunoinformatics. Acta Microbiol Immunol Hung 2014; 61(3):285-307.
31. Sefid F, Rasooli I, Jahangiri A, Bazmara H. Functional exposed amino acids of BauA as potential immunogen against Acinetobacter baumannii. Acta Biotheor 2015; 63(2):129-49.
32. Lu Y, Wei Y-q, Tian L, Zhao X, Yang L, Hu B, et al. Immunogene therapy of tumors with vaccine based on xenogeneic epidermal growth factor receptor. J Immunol 2003; 170(6):3162-70.
33. Ren S-x, Ren Z-j, Zhao M-y, Wang X-b, Zuo S-g, Yu F. Antitumor activity of endogenous mFlt4 displayed on a T4 phage nanoparticle surface. Acta Pharmacol Sin 2009; 30(5):637-45.
34. Ren S, Zuo S, Zhao M, Wang X, Wang X, Chen Y, et al. Inhibition of tumor angiogenesis in lung cancer by T4 phage surface displaying mVEGFR2 vaccine. Vaccine 2011; 29(34):5802-11.
35. Modjtahedi H, Komurasaki T, Toyoda H, Dean C. Anti-EGFR monoclonal antibodies which act as EGF, TGFa, HB-EGF and BTC antagonists block the binding of epiregulin to EGFR-expressing tumours. Int J Cancer 1998; 75(2):310-6.