title: Modeling Pulmonary Disease Pathways Using Recombinant Adeno-Associated Virus 6.2.
abstract: Viral vectors have been applied successfully to generate disease-related animal models and to functionally characterize target genes in vivo. However, broader application is still limited by complex vector production, biosafety requirements, and vector-mediated immunogenic responses, possibly interfering with disease-relevant pathways. Here, we describe adeno-associated virus (AAV) variant 6.2 as an ideal vector for lung delivery in mice, overcoming most of the aforementioned limitations. In a proof-of-concept study using AAV6.2 vectors expressing IL-13 and transforming growth factor-β1 (TGF-β1), we were able to induce hallmarks of severe asthma and pulmonary fibrosis, respectively. Phenotypic characterization and deep sequencing analysis of the AAV-IL-13 asthma model revealed a characteristic disease signature. Furthermore, suitability of the model for compound testing was also demonstrated by pharmacological intervention studies using an anti-IL-13 antibody and dexamethasone. Similarly, the AAV-TGF-β1 fibrosis model showed several disease-like pathophenotypes monitored by micro-computed tomography imaging and lung function measurement. Most importantly, analyses using stuffer control vectors demonstrated that in contrast to a common adenovirus-5 vector, AAV6.2 vectors did not induce any measurable inflammation and therefore carry a lower risk of altering relevant readouts. In conclusion, we propose AAV6.2 as an ideal vector system for the functional characterization of target genes in the context of pulmonary diseases in mice.
authors.affiliation: 1 Target Discovery Research.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Strobel
authors.affiliation: 2 Respiratory Diseases Research.
authors.forename: Matthias J
authors.initials: MJ
authors.lastname: Duechs
authors.affiliation: 3 Drug Metabolism and Pharmacokinetics, and.
authors.forename: Ramona
authors.initials: R
authors.lastname: Schmid
authors.affiliation: 1 Target Discovery Research.
authors.forename: Birgit E
authors.initials: BE
authors.lastname: Stierstorfer
authors.affiliation: 2 Respiratory Diseases Research.
authors.forename: Hannes
authors.initials: H
authors.lastname: Bucher
authors.affiliation: 1 Target Discovery Research.
authors.forename: Karsten
authors.initials: K
authors.lastname: Quast
authors.affiliation: 1 Target Discovery Research.
authors.forename: Detlef
authors.initials: D
authors.lastname: Stiller
authors.affiliation: 1 Target Discovery Research.
authors.forename: Tobias
authors.initials: T
authors.lastname: Hildebrandt
authors.affiliation: 1 Target Discovery Research.
authors.forename: Detlev
authors.initials: D
authors.lastname: Mennerich
authors.affiliation: 4 Translational Medicine and Clinical Pharmacology, Boehringer Ingelheim Pharma GmbH and Co. KG, Biberach an der Riss, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Gantner
authors.affiliation: 2 Respiratory Diseases Research.
authors.forename: Klaus J
authors.initials: KJ
authors.lastname: Erb
authors.affiliation: 2 Respiratory Diseases Research.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kreuz
mesh_terms: D000818:Animals; D001249:Asthma / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D004195:Disease Models, Animal; D005260:Female; D005822:Genetic Vectors; D054990:Idiopathic Pulmonary Fibrosis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018793:Interleukin-13 / Q000096:biosynthesis / Q000235:genetics; D008807:Mice, Inbred BALB C; D014161:Transduction, Genetic; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis / Q000235:genetics
chemical_list: D018793:Interleukin-13; C508961:Tgfb1 protein, mouse; D053773:Transforming Growth Factor beta1
keywords: IL-13; TGF-β1; animal models; asthma; lung fibrosis