title: Noninvasive Imaging Reveals Stable Transgene Expression in Mouse Airways After Delivery of a Nonintegrating Recombinant Adeno-Associated Viral Vector.
abstract: Gene therapy holds promise to cure a wide range of genetic and acquired diseases. Recent successes in recombinant adeno-associated viral vector (rAAV)-based gene therapy in the clinic for hereditary disorders such as Leber's congenital amaurosis and hemophilia B encouraged us to reexplore an rAAV approach for pulmonary gene transfer. Only limited clinical successes have been achieved for airway gene transfer so far, underscoring the need for further preclinical development of rAAV-based gene therapy for pulmonary disorders. We sought to determine the preclinical potential of an airway-tropic serotype, rAAV2/5, encoding reporter genes when delivered to mouse airways. Although several groups have assessed the stability of gene transfer using a nonintegrating rAAV in mouse airways, long-term stability for more than a year has not been reported. Additionally, an extensive quantitative analysis of the specific cell types targeted by rAAV2/5 using cell-specific markers is lacking. We obtained sustained gene expression in upper and lower airways up to 15 months after vector administration, a substantial proportion of the lifespan of a laboratory mouse. In addition, we demonstrated that readministration of rAAV2/5 to the airways is feasible and increases gene expression 14 months after primary vector administration, despite the presence of circulating neutralizing antibodies. Finally, identification of transduced cell types revealed different subpopulations being targeted by rAAV2/5, with 64% of β-galactosidase-positive cells being ciliated cells, 34% club cells in the conducting airways, and 75% alveolar type II cells in the alveoli at 1 month postinjection. This underscores the therapeutic potential of a nonintegrating rAAV vector to develop a gene therapeutic drug for a variety of pulmonary disorders, such as cystic fibrosis, primary ciliary dyskinesia, and surfactant deficiencies.
authors.affiliation: 1 Laboratory for Molecular Virology and Gene Therapy , Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Flanders, Belgium.
authors.forename: Dragana
authors.initials: D
authors.lastname: Vidović
authors.affiliation: 1 Laboratory for Molecular Virology and Gene Therapy , Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Flanders, Belgium.|2 Leuven Viral Vector Core, KU Leuven, Flanders, Belgium.
authors.forename: Rik
authors.initials: R
authors.lastname: Gijsbers
authors.affiliation: 1 Laboratory for Molecular Virology and Gene Therapy , Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Flanders, Belgium.
authors.forename: Ana
authors.initials: A
authors.lastname: Quiles-Jimenez
authors.affiliation: 3 Department of Microbiology and Immunology, KU Leuven, Flanders, Belgium.|4 Vlaams Instituut Voor Biotechnologie, Translational Immunology Laboratory, Flanders, Belgium .
authors.forename: James
authors.initials: J
authors.lastname: Dooley
authors.affiliation: 2 Leuven Viral Vector Core, KU Leuven, Flanders, Belgium.|5 Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Flanders, Belgium.
authors.forename: Chris
authors.initials: C
authors.lastname: Van den Haute
authors.affiliation: 5 Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Flanders, Belgium.
authors.forename: Anke
authors.initials: A
authors.lastname: Van der Perren
authors.affiliation: 3 Department of Microbiology and Immunology, KU Leuven, Flanders, Belgium.|4 Vlaams Instituut Voor Biotechnologie, Translational Immunology Laboratory, Flanders, Belgium .
authors.forename: Adrian
authors.initials: A
authors.lastname: Liston
authors.affiliation: 5 Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Flanders, Belgium.
authors.forename: Veerle
authors.initials: V
authors.lastname: Baekelandt
authors.affiliation: 1 Laboratory for Molecular Virology and Gene Therapy , Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Flanders, Belgium.
authors.forename: Zeger
authors.initials: Z
authors.lastname: Debyser
authors.affiliation: 1 Laboratory for Molecular Virology and Gene Therapy , Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Flanders, Belgium.
authors.forename: Marianne Sylvia
authors.initials: MS
authors.lastname: Carlon
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics* / Q000302:isolation & purification; D005786:Gene Expression Regulation; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008168:Lung / Q000378:metabolism* / Q000473:pathology; D008171:Lung Diseases / Q000235:genetics / Q000628:therapy; D051379:Mice; D057054:Molecular Imaging; D019076:Transgenes / Q000235:genetics