title: Inhalation of nebulized perfluorochemical enhances recombinant adenovirus and adeno-associated virus-mediated gene expression in lung epithelium.
abstract: Use of perfluorochemical liquids during intratracheal vector administration enhances recombinant adenovirus and adeno-associated virus (AAV)-mediated lung epithelial gene expression. We hypothesized that inhalation of nebulized perfluorochemical vapor would also enhance epithelial gene expression after subsequent intratracheal vector administration. Freely breathing adult C57BL/6 mice were exposed for selected times to nebulized perflubron or sterile saline in a sealed Plexiglas chamber. Recombinant adenoviral vector was administered by transtracheal puncture at selected times afterward and mice were killed 3 days after vector administration to assess transgene expression. Mice tolerated the nebulized perflubron without obvious ill effects. Vector administration 6 hr after nebulized perflubron exposure resulted in an average 540% increase in gene expression in airway and alveolar epithelium, compared with that with vector alone or saline plus vector control (p<0.05). However, vector administration 1 hr, 1 day, or 3 days after perflubron exposure was not different from either nebulized saline with vector or vector alone and a 60-min exposure to nebulized perflubron is required. In parallel pilot studies in macaques, inhalation of nebulized perflubron enhanced recombinant AAV2/5 vector expression throughout the lung. Serial chest radiographs, bronchoalveolar lavages, and results of complete blood counts and serum biochemistries demonstrated no obvious adverse effects of nebulized perflubron. Further, one macaque receiving nebulized perflubron only was monitored for 1 year with no obvious adverse effects of exposure. These results demonstrate that inhalation of nebulized perflubron, a simple, clinically more feasible technique than intratracheal administration of liquid perflubron, safely enhances lung gene expression.
authors.affiliation: Pulmonary and Critical Care, Vermont Lung Center, University of Vermont College of Medicine, Burlington, VT 05405, USA.
authors.forename: Travis
authors.initials: T
authors.lastname: Beckett
authors.forename: Laura
authors.initials: L
authors.lastname: Bonneau
authors.forename: Alan
authors.initials: A
authors.lastname: Howard
authors.forename: James
authors.initials: J
authors.lastname: Blanchard
authors.forename: Juan
authors.initials: J
authors.lastname: Borda
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Weiner
authors.forename: Lili
authors.initials: L
authors.lastname: Wang
authors.forename: Guang Ping
authors.initials: GP
authors.lastname: Gao
authors.forename: Jay K
authors.initials: JK
authors.lastname: Kolls
authors.forename: Rudolf
authors.initials: R
authors.lastname: Bohm
authors.forename: Denny
authors.initials: D
authors.lastname: Liggitt
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Weiss
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000280:Administration, Inhalation; D000704:Analysis of Variance; D000818:Animals; D018893:Bronchoalveolar Lavage; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D005466:Fluorocarbons / Q000008:administration & dosage / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006842:Hydrocarbons, Brominated; D008251:Macaca; D051379:Mice; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000378:metabolism*; D014057:Tomography, X-Ray Computed; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D005466:Fluorocarbons; D006842:Hydrocarbons, Brominated; D001616:beta-Galactosidase; C003072:perflubron