Mohammad KHALAJ-KONDORI, Mahboobeh KOVOOSI, Mohammad RAHMATI-YAMCHI, Mehdi KADIVAR

Preparation of a transferrin-targeted M13-based gene nanocarrier and evaluation of its efficacy for gene delivery and expression in eukaryote cells

Bacteriophages are appropriate gene carriers that might be targeted toward target cells using different strategies. Here we prepared a transferrin-targeted M13-based gene nanocarrier (Tf-targeted M13-GFP) and examined its gene delivery and expression efficacy in the AGS cell line. M13 phagemid particles bearing a GFP expression cassette (M13-GFP) were obtained from a recombinant lambda phage through an in vivo excision procedure. Chemical coupling of human holotransferrin molecules (Tf) to the surface of these phagemid particles resulted in Tf-targeted M13-GFP formation, which was then characterized by Phage-ELISA and Cell-ELISA experiments. Immunocytochemistry (ICC) and fluorescence-activated cell sorting (FACS) analysis were used for internalization assay and examination of gene delivery/expression efficacies in the human AGS cell line. The ELISA experiments revealed high-density attachment of Tf molecules to the surface of M13-GFP particles and ICC confirmed highly efficient internalization of the Tf-targeted M13-GFP particles into the AGS cells. Moreover, FACS analysis showed significant increase of GFP-positive cell counts in the samples treated with Tf-targeted M13-GFP (8.09%) in comparison with the samples treated with wild M13-GFP (1.2%). We conclude that this strategy might improve phage-mediated gene delivery and expression in eukaryote cells.

PDF

___

Abedheydari E, Khalaj-Kondori M, Hosseinpour-Faizi MA, Kosari-Nasab M (2014). A comparative investigation on efficiency of bacteriophage lambda and M13 based vectors for delivering and expression of transgene in eukaryote cells. Journal of Cell and Molecular Research 6: 70-76.
Bakhshinejad B, Karimi M, Sadeghizadeh M (2014). Bacteriophages and medical oncology: targeted gene therapy of cancer. Med Oncol 31: 110.
Bakhshinejad B, Sadeghizadeh M (2014). Bacteriophages as vehicles for gene delivery into mammalian cells: prospects and problems. Expert Opin Drug Deliv 11: 1561-1574.
Chung YS, Sabel K, Krönke M, Klimka A (2008). Gene transfer of Hodgkin cell lines via multivalent anti-CD30 scFv displaying bacteriophage. BMC Mol Biol 9: 37.
Eguchi A, Furusawa H, Yamamoto A, Akuta T, Hasegawa M, Okahata Y, Nakanishi M (2005). Optimization of nuclear localization signal for nuclear transport of DNA-encapsulating particles. J Control Release 104: 507-519.
Gupta A, Onda M, Pastan I, Adhya S, Chaudhary VK (2003). High-density functional display of proteins on bacteriophage lambda. J Mol Biol 334: 241-254.
Hodyra K, Dąbrowska K (2015). Molecular and chemical engineering of bacteriophages for potential medical applications. Arch Immunol Ther Exp (Warsz) 63: 117-127.
Huang RK, Steinmetz NF, Fu CY, Manchester M, Johnson JE (2011). Transferrin-mediated targeting of bacteriophage HK97 nanoparticles into tumor cells. Nanomedicine 6: 55-68.
Ishida O, Maruyama K, Tanahashi H, Iwatsuru M, Sasaki K, Eriguchi M, Yanagie H (2001). Liposomes bearing polyethyleneglycol-coupled transferrin with intracellular targeting property to the solid tumors in vivo. Pharm Res 18: 1042-1048.
Ivanenkov VV, Felici F, Menon AG (1999a). Targeted delivery of multivalent phage display vectors into mammalian cells. Biochim Biophys Acta 1448: 463-472.
Ivanenkov V, Felici F, Menon AG (1999b). Uptake and intracellular fate of phage display vectors in mammalian cells. Biochim Biophys Acta 1448: 450-462.
Khalaj-Kondori M, Sadeghizadeh M, Behmanesh M, Saggio I, Monaci P (2011). Chemical coupling as a potent strategy for preparation of targeted bacteriophage-derived gene nanocarriers into eukaryotic cells. J Gene Med 13: 622-631.
Khalaj-Kondori M, Sadeghizadeh M, Behmanesh M (2010). Bacteriophage lambda-mediated gene transfer into human AGS cell line. In: Proceedings of the 2nd International Conference on Chemical, Biological and Environmental Engineering (ICBEE); Cairo, Egypt. New York, NY, USA: IEEE, pp. 176-179.
Kim A, Shin TH, Shin SM, Pham CD, Choi DK, Kwon MH, Kim YS (2012). Cellular internalization mechanism and intracellular trafficking of filamentous M13 phages displaying a cell-penetrating transbody and TAT peptide. PLoS One 7: e51813.
Lai H, Nakase I, Lacoste E, Singh NP, Sasaki T (2009). Artemisinin-transferrin conjugate retards growth of breast tumors in the rat. Anticancer Res 29: 3807-3810.
Lai H, Sasaki T, Singh NP, Messay A (2005). Effects of artemisinin-tagged holotransferrin on cancer cells. Life Sci 76: 1267-1279.
Li K, Chen Y, Li S, Nguyen HG, Niu Z, You S, Mello CM, Lu X, Wang Q (2010). Chemical modification of M13 bacteriophage and its application in cancer cell imaging. Bioconjugate Chem 21: 1369-1377.
Li Z, Zhang J, Zhao R, Xu Y, Gu J (2005). Preparation of peptide-targeted phagemid particle using a protein III-modified helper phage. Biotechniques 39: 493-497.
Mount JD, Samoylova TI, Morrison NE, Cox NR, Baker HJ, Petrenko VA (2004). Cell targeted phagemid rescued by preselected landscape phage. Gene 341: 59-65.
Nakase I, Gallis B, Takatani-Nakase T, Oh S, Lacoste E, Singh NP, Goodlett DR, Tanaka S, Futaki S, Lai H et al. (2009). Transferrin receptor-dependent cytotoxicity of artemisinin-transferrin conjugates on prostate cancer cells and induction of apoptosis. Cancer Lett 274: 290-298.
Poul MA, Marks JD (1999). Targeted gene delivery to mammalian cells by filamentous Bacteriophage. J Mol Biol 288: 203-211.
Qian ZM, Li H, Sun H, Ho K (2002). Targeted drug delivery via the transferrin receptor-mediated endocytosis pathway. Pharmacol Rev 54: 561-587.
Shoae-Hassani A, Keyhanvar P, Seifalian AM, Mortazavi-Tabatabaei SA, Ghaderi N, Issazadeh K, Amirmozafari N, Verdi J (2013a). λ Phage nanobioparticle expressing apoptin efficiently suppress human breast carcinoma tumor growth in vivo. PLoS One 8: e79907.
Shoae-Hassani A, Mortazavi-Tabatabaei SA, Sharif S, Madadi S, Rezaei-Khaligh H, Verdi J (2013b) Recombinant λ bacteriophage displaying nanobody towards third domain of HER-2 epitope inhibits proliferation of breast carcinoma SKBR-3 cell line. Arch Immunol Ther Exp (Warsz) 61: 75-83.
Singh P, Destito G, Schneemann A, Manchester M (2006). Canine parvovirus-like particles, a novel nanomaterial for tumour targeting. J Nanobiotechnol 4: 2.
Stephanopoulos N, Tong GJ, Hsiao SC, Francis MB (2010). Dual-surface modified virus capsids for targeted delivery of photodynamic agents to cancer cells. ACS Nano 4: 6014-6020.
Urbanelli L, Ronchini C, Fontana L, Menard S, Orlandi R, Monaci P (2001). Targeted gene transduction of mammalian cells expressing the HER2/neu receptor by filamentous phage. J Mol Biol 313: 965-976.
Volcy K, Dewhurst S (2009). Proteasome inhibitors enhance bacteriophage lambda mediated gene transfer in mammalian cells. Virology 384: 77-87.