Enhanced gene delivery to human airway epithelial cells using an integrin-targeting lipoplex

Background Current liposome-based delivery methods for cystic fibrosis (CF) gene therapy are limited by their poor efficiencies. One way to improve this is to use a receptor/ligand interaction to increase binding of the transfection complex with the target cell. Methods and results We have tested a synthetic peptide containing an alpha (v) integrin-binding motif (arginine-glycine-aspartic acid, RGD) and a DNA-binding domain (polylysine) for enhancement of liposome-mediated gene delivery. We have shown that integrin proteins capable of binding the RGD motif are located on the apical surface of a polarized human bronchial epithelial cell:line (16HBE). Luciferase gene transfer efficiency to subconfluent 16HBE cells was 10-200 times higher than gene transfer using either liposome or peptide alone. This peptide-mediated enhancement was observed at all cellular contact times including those as short as 1 min. Although the transfection efficiency is reduced when the 16HBE cells are grown as polarized monolayers, peptide-mediated enhancement of lipofection is maintained. Transfection with a lipopolyplex containing an RGE (arginine-glucine-glutamic acid) control peptide that cannot bind to the alpha (v) integrin molecules, or competitive inhibition with antibodies against RGD-binding integrins, reduced gene transfer. Confocal microscopy indicated that the peptide increased plasmid delivery to the cell via receptor-mediated endocytosis. Conclusion These results indicate that integrin-binding peptides represent one way to enhance liposome-mediated gene delivery to pulmonary epithelia. Copyright (C) 2001 John Wiley & Sons, Ltd.