LIPOSOMES ENHANCE DELIVERY AND EXPRESSION OF AN RGD-OLIGOLYSINE GENE-TRANSFER VECTOR IN HUMAN TRACHEAL CELLS

Nonviral gene delivery systems consist predominantly of lipoplexes or receptor-targeting and nontargeting polyplexes. We examined integrin-m ediated gene delivery using an Arg-Gly-Asp/oligo-L-lysine ([K](16)RGD) cyclic peptide and investigated its gene transfer efficiency when ass ociated with a cationic liposome. We demonstrated that human cystic fi brosis and noncystic fibrosis tracheal epithelial cells in culture exp ress integrins that recognise the RGD integrin-binding motif. We found a 10-fold (P<0.01) increased expression of a luciferase encoding plas mid in these cells when complexing the plasmid to the [K](16)RGD pepti de as compared with plasmid alone. This increase was specific to the [ K](16)RGD peptide since neither a [K](16)RGE nor a [K](16) peptide gav e a comparable increase. Expression was further enhanced 30-fold (P<0. 01) with lipofectamine and the ratio of DNA/peptide/lipofectamine was critical for specificity and expression. Fluorescence and radioactive labelling of the complex showed that the [K](16)RGD peptide increased the endocytic uptake of DNA into cells. The cell association of both D NA and peptide increased even further with lipofectamine. Confocal mic roscopy showed that the [K](16)RGD peptide and the DNA internalised to gether within 30 min and localised to vesicles in the perinuclear regi on. These results show that an integrin-binding ligand can deliver gen etic material to airway cells and that a cationic liposome can enhance the efficacy of this nonviral vector system.