Enhanced gene delivery to human airway epithelial cells using an integrin‐targeting lipoplex
Identifieur interne : 002387 ( Main/Exploration ); précédent : 002386; suivant : 002388Enhanced gene delivery to human airway epithelial cells using an integrin‐targeting lipoplex
Auteurs : Emily S. Scott [Royaume-Uni] ; John W. Wiseman [Royaume-Uni] ; Martin J. Evans [Royaume-Uni] ; William H. Colledge [Royaume-Uni]Source :
- The Journal of Gene Medicine [ 1099-498X ] ; 2001-03.
English descriptors
- Teeft :
- Adhesion, Airway, Airway epithelial cells, Airway epithelium, Apical, Apical surface, Assay, Biol chem, Bronchial epithelial cells, Bronectin, Brosis, Cationic, Cationic liposome, Cell adhesion, Cell adhesion assay, Cell lysates, Cell surface receptors, Collagen substrate, Confocal microscopy, Contact time, Control peptide, Copyright, Cyclized peptide, Cystic, Data points, Endocytosis, Enhancement, Epithelial, Epithelial cell line, Epithelial cells, Epithelium, Gene, Gene delivery, Gene therapy, Gene transfer, Gene transfer efficiency, Hours cells, Integrin, Integrin receptors, Integrin subunit, Integrins, John wiley sons, Linear peptide, Lipofection, Lipoplex, Lipopolyplex, Liposome, Luciferase, Luciferase activity, Nasal epithelium, Pcmvluc, Peptide, Plasmid, Present study, Primary antibody, Protein content, Receptor, Reporter gene delivery, Room temperature, Secondary antibody, Sigma, Subunit, Transfected, Transfection, Undifferentiated, Values differences.
Abstract
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 αv 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 αv 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 © 2001 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/jgm.172
Affiliations:
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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 αv 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 αv 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 © 2001 John Wiley & Sons, Ltd.</div></front></TEI><affiliations><list><country><li>Royaume-Uni</li></country><region><li>Angleterre</li><li>Angleterre de l'Est</li></region><settlement><li>Cambridge</li></settlement><orgName><li>Université de Cambridge</li></orgName></list><tree><country name="Royaume-Uni"><region name="Angleterre"><name sortKey="Scott, Emily S" sort="Scott, Emily S" uniqKey="Scott E" first="Emily S." last="Scott">Emily S. 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