Enhanced in vivo airway gene transfer via transient modification of host barrier properties with a surface-active agent.
Effective adenoviral gene therapy requires efficient viral vector entry into epithelial cells. Injured airway epithelia display enhanced gene transfer, reflecting in part increased vector access to protected cell populations and/or protected basolateral membranes. We tested whether adenoviral gene transfer is enhanced by modification of the epithelial barrier in mouse nasal airways with a nonionic detergent (polidocanol, PDOC). In C57BL/6 mice, 1.6 x 10(9) PFU of Ad5CMV LacZ (AdLacZ) instilled into the right nostril produced negligible gene transfer to the nasal epithelium 2 days after dosing, but significant, dose-dependent increases in gene transfer were achieved by pretreatment with PDOC. Permeation of the electron-dense tracer lanthanum into the intercellular junctions of PDOC (0.1%)-treated murine nasal epithelium, but not into intercellular junctions of vehicle controls, is consistent with PDOC-mediated increases in tight junctional permeability. In CF(-/-) mice, significant gene expression was not detectable after exposure to Ad5CBCFTR alone (1.4 x 10(9) PFU in 20 microl; AdCFTR), but PDOC pretreatment prior to AdCFTR instillation produced functional expression of CFTR (measured as deltaPD) 5 days after instillation. Because the development and testing of lung gene therapy will principally occur in children and adults with airway disease, AdLacZ gene transfer with and without PDOC pretreatment was examined in infected nasal airways. Gene expression was significantly reduced in infected as compared with uninfected airways. We conclude that the use of adjuvant surface-active and/or membrane-perturbing agents, synthetic or naturally derived, may provide a novel approach to enhancing the efficiency of adenoviral gene transfer.
Parsons
D W
DW
Cystic Fibrosis Center, Department of Medicine, University of North Carolina at Chapel Hill, 27599, USA. parsonsd@wch.sa.gov.au
Grubb
B R
BR
Johnson
L G
LG
Boucher
R C
RC
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects / Q000648:ultrastructure; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D007365:Intercellular Junctions / Q000648:ultrastructure; D007763:Lac Operon; D051379:Mice; D009297:Nasal Mucosa / Q000166:cytology / Q000187:drug effects*; D009666:Nose / Q000382:microbiology; D010539:Permeability / Q000187:drug effects; D000077423:Polidocanol; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011552:Pseudomonas Infections; D011550:Pseudomonas aeruginosa
D003902:Detergents; D000077423:Polidocanol; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D011092:Polyethylene Glycols