Transgene expression in mouse airway epithelium by aerosol gene therapy with PEI-DNA complexes.
Gene therapy targeted at the respiratory epithelium holds therapeutic potential for diseases such as cystic fibrosis and alpha-1 anti-trypsin deficiency. A variety of approaches such as intranasal or intratracheal instillation and aerosol delivery have been utilized to target genes to the airways. Polyethylenimine (PEI), a linear or branched polycationic polymer, has been used for delivery of genes to various organs. In this study, using fluorescein isothiocyanate (FITC)-labeled branched PEI, we initially examined the localization of PEI in the lungs after aerosol delivery to Balb/C mice. Further, after aerosol delivery of PEI-CAT DNA, in situ immunostaining for chloramphenicol acetyl transferase (CAT) protein was used to localize the transgene expression within the lungs. Immunohistochemistry for CAT, as well as localization of FITC-labeled PEI, revealed that after aerosol delivery, the PEI-DNA complexes deposit and subsequently transfect most of the epithelial cells in the conducting airways (including the peripheral airways). High levels of CAT were detected at 24 h after aerosol exposure and significant CAT expression was detected in the lungs up to 28 days after a single aerosol exposure. The data suggest that aerosol delivery of PEI-DNA complexes could be effective for the treatment of pulmonary diseases such as cystic fibrosis and alpha-1 anti-trypsin deficiency.
Gautam
A
A
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas 77030, USA.
Densmore
C L
CL
Golunski
E
E
Xu
B
B
Waldrep
J C
JC
D000280:Administration, Inhalation; D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D003550:Cystic Fibrosis / Q000628:therapy; D016503:Drug Delivery Systems; D004847:Epithelial Cells / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism*; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000494:pharmacology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008168:Lung / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D011094:Polyethyleneimine / Q000627:therapeutic use*; D013997:Time Factors; D014162:Transfection; D019076:Transgenes; D019896:alpha 1-Antitrypsin Deficiency / Q000628:therapy
D005456:Fluorescent Dyes; D011094:Polyethyleneimine; D015500:Chloramphenicol O-Acetyltransferase; D016650:Fluorescein-5-isothiocyanate