title: Perfluorochemical liquid-enhanced adenoviral vector distribution and expression in lungs of spontaneously breathing rodents.
abstract: Perfluorochemical (PFC) liquids have been shown to improve gas exchange and lung compliance in models of lung injury. We reasoned they may also be useful as a vehicle for gene transfer by improving transgene distribution throughout the lung as well as increasing total transgene expression. We have developed a model for PFC liquid use in spontaneously breathing rodents that obviates the need for intubation and ventilation. Intratracheal instillation of the adenoviral vector Adlac-Z resulted in patchy distribution of beta-galactosidase (beta-gal) activity as demonstrated using X-gal histochemistry. In contrast, in rats instilled with Adlac-Z followed by instillation of PFC liquid, more uniformly distributed and increased beta-gal activity was observed. Activity in distal airway and alveolar epithelium was particularly increased. Quantitative measure of beta-gal activity in lung homogenates demonstrated a 3- to 6-fold increase in total activity in lungs of rats receiving Adlac-Z and PFC liquid compared to animals receiving Adlac-Z alone. These studies show that PFC liquids can enhance both the distribution and the total amount of transgene expressed following adenoviral-mediated vector transfer to lungs during spontaneous breathing. Use of PFC liquids may increase the efficacy of gene transfer strategies for treatment of cystic fibrosis and other lung diseases.
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Fred Hutchinson Cancer Research Center, University of Washington, Seattle 98109-10,USA.dweiss@fhcrc.org
authors.forename: D J
authors.initials: DJ
authors.lastname: Weiss
authors.forename: T P
authors.initials: TP
authors.lastname: Strandjord
authors.forename: J C
authors.initials: JC
authors.lastname: Jackson
authors.forename: J G
authors.initials: JG
authors.lastname: Clark
authors.forename: D
authors.initials: D
authors.lastname: Liggitt
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D003673:Defective Viruses; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D005466:Fluorocarbons / Q000008:administration & dosage / Q000494:pharmacology*; D005663:Furans / Q000008:administration & dosage / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007763:Lac Operon / Q000235:genetics; D008168:Lung / Q000000981:diagnostic imaging / Q000201:enzymology* / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011859:Radiography; D051381:Rats; D017207:Rats, Sprague-Dawley; D012119:Respiration; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
chemical_list: D005466:Fluorocarbons; D005663:Furans; C073032:perfluorobutyl tetrahydrofuran; D001616:beta-Galactosidase