A RGD-containing oligopeptide (K)₁₆GRGDSPC: A novel vector for integrin-mediated targeted gene delivery

Abstract

A 23 amino acid, bifunctional integrin-targeted synthetic oligopeptide was evaluated for ex vivo gene delivery to rabbit bone marrow stromal cells (BMSCs). Synthesis of the peptide (K)₁₆GRGDSPC was performed on a solid-phase batch peptide synthesizer. BMSCs were transfected with plasmid DNA coding for luciferase by (K)₁₆GRGDSPC and the transfection efficiency was assayed. The influences of chloroquine and polyethyleneimine on the transfection efficiency were also examined. The target specificity of (K)₁₆GRGDSPC to mediate exogenous gene into BMSCs was analyzed using cell attachment test and gene delivery inhibition test. The results showed that the transfection efficiency of the oligopeptide vector was lower than that of Lipofectamine. But in the presence of endosomal buffer chloroquine or endosomal disrupting agent polyethyleneimine, the transfection efficiency of the vector was greatly enhanced. In addition, RGD-containing peptides inhibited BMSCs’ attachment to the 96-well plates pretreated with fibronectin or vitronectin and significantly decreased the transfection efficiency of the oligopeptide vector. These studies demonstrated that oligopeptide (K)₁₆GRGDSPC was an ideal novel targeted non-viral gene delivery vector, which was easy to be synthesized, high efficient and low cytotoxicity. The vector could effectively deliver exogenous gene into rat BMSCs.

Keywords

targeted non-viral vector / RGD-containing peptide / gene delivery

References

[1] RichardP., Harbottle, RobertG., et al.. An RGD-oligolysine peptide: a prototype construct for interin-mediated gene delivery. Human Gene Therapy, 1998, 9: 1037-1047

[2] WangY. T., ZhengQ. X., GuoX. D., et al.. Optimal isolation and identification of rat autogenous bone marrow derived mesenchymal stem cells. J Huazhong Univ Sci Tech (Health Sci) (Chinese), 2003, 32: 526-529

[3] SiTuZ. Q., WuJ. Z.. Cell Culture (Chinese), 2003, Beijing, World Book Printing Corporation: 137

[4] ChowdhuryN. R., WuC. H., WuG. Y., et al.. Fate of DNA targeted to the liver by asialoglyco-protein receptor-mediated endocytosis in vivo. J Biol Chem, 1993, 268: 11 265-11 271

[5] WagenerE., ZenkeM., CottonM., et al.. Transferrin-polycation conjugates as carriers for DNA uptake into cells. Proc Natl Acad Sci USA, 1991, 87: 3410-3414

[6] KlinkD., YuQ. C., GlickM. C., et al.. Lactosylated poly-L-lysine targets a potential lactose receptor in cystic fibrosis and non-cystic fibrosis airway epithelial cells. Mol Ther, 2003, 7: 73-80

[7] KawakamiS., HattoriY., LuY., et al.. Effect of cationic charge on receptor-mediated transfection using mannosylated cationic liposome/plasmid DNA complexes following the intravenous administration in mice. Pharmazie, 2004, 59(5): 405-408

[8] CollinsL., FabreJ. W.. A synthetic peptide vector system for optimal gene delivery to corneal endothelium. J Gene Med, 2004, 6: 185-194

[9] VerrierS., PalluS., BareilleR., et al.. Function of linear and cyclic RGD-containing peptides in osteoprogenitor cells adhesion process. Biomaterials, 2002, 23: 585