title: Intraluminal water increases expression of plasmid DNA in rat lung.
abstract: Effective gene delivery to specific organs is a major goal for human gene therapy. The lung's structure allows instillation of agents into the airspaces, directly adjacent to the lung epithelium. We hypothesized that the airspace instillation of hypotonic solutions would increase the permeability of the lung epithelium and increase DNA uptake. This hypothesis was tested by instilling plasmid DNA (p4241) encoding the luciferase gene in isotonic and hypotonic solutions. The highest luciferase expression in the lung was achieved after the instillation of this plasmid DNA in distilled water. Aerosolization of water just before the instillation of the plasmid DNA also enhanced the expression level of luciferase in the lung. In addition, an intralobar instillation of the plasmid DNA in water significantly increased the luciferase expression, suggesting that the instillation of the plasmid over a smaller surface area increased expression. Levels of expression could be measured for 3 days. Water increases the permeability of lung epithelial cells transiently and/or enhances gene expression and can be used to achieve gene expression in the lung airspaces for short intervals without toxicity.
authors.affiliation: Department of Anesthesia and Medicine, University of California, San Francisco 94143, USA.
authors.forename: T
authors.initials: T
authors.lastname: Sawa
authors.forename: H
authors.initials: H
authors.lastname: Miyazaki
authors.forename: J F
authors.initials: JF
authors.lastname: Pittet
authors.forename: J H
authors.initials: JH
authors.lastname: Widdicombe
authors.forename: M A
authors.initials: MA
authors.lastname: Gropper
authors.forename: S
authors.initials: S
authors.lastname: Hashimoto
authors.forename: D J
authors.initials: DJ
authors.lastname: Conrad
authors.forename: H G
authors.initials: HG
authors.lastname: Folkesson
authors.forename: R
authors.initials: R
authors.lastname: Debs
authors.forename: J R
authors.initials: JR
authors.lastname: Forsayeth
authors.forename: B
authors.initials: B
authors.lastname: Fox
authors.forename: J P
authors.initials: JP
authors.lastname: Wiener-Kronish
mesh_terms: D000818:Animals; D001834:Body Water; D002463:Cell Membrane Permeability; D004274:DNA, Recombinant / Q000235:genetics*; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D004274:DNA, Recombinant