title: Efficient gene transfer into the mouse lung by fetal intratracheal injection of rAAV2/6.2.
abstract: Fetal gene therapy is one of the possible new therapeutic strategies for congenital or perinatal diseases with high mortality or morbidity. We developed a novel delivery strategy to inject directly into the fetal mouse trachea. Intratracheal (i.t.) injection at embryonic day 18 (E18) was more efficient in targeting the fetal lung than conventional intra-amniotic (i.a.) delivery. Viral vectors derived from adeno-associated virus serotype 6.2, with tropism for the airway epithelium and not earlier tested in the fetal mouse lung, were injected into the fetal trachea. Bioluminescence (BL) imaging (BLI) was combined with magnetic resonance (MR) imaging (MRI) for noninvasive and accurate localization of transgene expression in vivo. Histological analysis for β-galactosidase (β-gal) revealed 17.5% of epithelial cells transduced in the conducting airways and 1.5% in the alveolar cells. Stable gene expression was observed up to 1 month after injection. This study demonstrates that direct injection of rAAV2/6.2 in the fetal mouse trachea is superior to i.a. delivery for transducing the lung. Second, as stable gene transfer was detected up to 1 postnatal month, this approach may be useful to evaluate fetal gene therapy for pulmonary diseases such as cystic fibrosis, requiring both substantial numbers of transduced cells as well as prolonged gene expression to obtain a stable phenotypic effect.
authors.affiliation: Department of Molecular and Cellular Medicine, Katholieke Universiteit Leuven, Flanders, Belgium.
authors.forename: Marianne
authors.initials: M
authors.lastname: Carlon
authors.forename: Jaan
authors.initials: J
authors.lastname: Toelen
authors.forename: Anke
authors.initials: A
authors.lastname: Van der Perren
authors.forename: Luk H
authors.initials: LH
authors.lastname: Vandenberghe
authors.forename: Veerle
authors.initials: V
authors.lastname: Reumers
authors.forename: Lourenço
authors.initials: L
authors.lastname: Sbragia
authors.forename: Rik
authors.initials: R
authors.lastname: Gijsbers
authors.forename: Veerle
authors.initials: V
authors.lastname: Baekelandt
authors.forename: Uwe
authors.initials: U
authors.lastname: Himmelreich
authors.forename: James M
authors.initials: JM
authors.lastname: Wilson
authors.forename: Jan
authors.initials: J
authors.lastname: Deprest
authors.forename: Zeger
authors.initials: Z
authors.lastname: Debyser
mesh_terms: D000653:Amniotic Fluid; D000818:Animals; D000229:Dependovirus / Q000235:genetics; D005333:Fetus; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D008168:Lung; D051379:Mice; D023421:Models, Animal; D014132:Trachea