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

✍ Scribed by Emily S. Scott; John W. Wiseman; Martin J. Evans; William H. Colledge

• Publisher: John Wiley and Sons
• Year: 2001
• Tongue: English
• Weight: 309 KB
• Volume: 3
• Category: Article
• ISSN: 1099-498X
• DOI: 10.1002/jgm.172

✦ Synopsis

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 alphav 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 alphav 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.