Abstract
Poster: 228Systematic mapping of AAV2.5-mediated gene transduction in the ferret lung
Journal of cystic fibrosis, Vol.25(Suppl 2), pp.117-117
09/2026
DOI: 10.1016/j.jcf.2026.07.259
Abstract
Effective gene therapy for cystic fibrosis (CF) and other pulmonary diseases requires efficient, cell-type-specific delivery of a functional CFTR gene. Ferrets are a highly relevant preclinical model for airway gene transfer because their airway anatomy and cellular composition closely resemble those of humans, and CF ferrets recapitulate key CF lung phenotypes. The rAAV2.5T vector shows enhanced apical tropism in human airway epithelia in vitro and can transduce ferret lungs in vivo after intratracheal dosing, but its epithelial cell-type tropism in adult ferret airways remains poorly defined. Here, we comprehensively profile cell-type-specific rAAV2.5T transduction in adult ferret lungs.
To minimize immune responses to transgene products, we used two immune-tolerant transgenic ferret models. OCT4-GFP ferrets, carrying an OCT4-IRES-eGFP knock-in allele silenced in the adult lung, received rAAV2.5-eGFP, and FOXI1-Cre::ROSA26-LoxPtdTomatoLoxPStop-GFP (Cre-ROSA-TG) ferrets received rAAV2.5T-Cre. Ferrets were dosed with 1 × 1013 vg/kg in 2 ml/kg inoculum containing 200 μM doxorubicin. Lungs were harvested three days post-transduction, and the trachea plus all lobes were analyzed by confocal immunofluorescence. GFP⁺ cells were co-localized with markers for ciliated cells (FOXJ1), club cells (SCGB1A1), basal cells (KRT5 or TP63), goblet cells (MUC5AC or MUC5B), ionocytes (BSND or CFTR), and alveolar type I (AGER) and II cells (SPC). Regional distribution and cell-type-specific transduction efficiencies were quantified in Fiji using the Analyze Particles function.
rAAV2.5 mediated widespread epithelial transduction throughout the ferret airway. In the trachea and large bronchi, ciliated, goblet, basal, and ionocyte populations were efficiently transduced. In mid-sized bronchioles, transduction was enriched in club cells with lower levels in basal cells. In terminal bronchioles and alveoli, GFP signal was predominantly localized to AT1 cells, with occasional AT2 labeling. Overall, rAAV2.5 showed broad epithelial coverage with preferential targeting of club cells and AT1 cells. rAAV2.5-Cre and rAAV2.5-GFP exhibited similar cell-type patterns, although Cre-mediated recombination was more sensitively detected in Cre-ROSA-TG lungs than GFP expression in OCT4-GFP lungs.
This work provides a cell-resolved atlas of rAAV2.5-mediated gene delivery across the ferret respiratory tree. The broad epithelial tropism from proximal airways to distal alveoli supports the translational potential of AAV2.5 vectors for CF and other diffuse lung diseases and underscores the utility of immune-tolerant ferret models for evaluating airway gene transfer with reduced immunologic confounders.
Details
- Title: Subtitle
- Poster: 228Systematic mapping of AAV2.5-mediated gene transduction in the ferret lung
- Creators
- L. Jiang - University of Alabama at BirminghamH. Dong - University of Alabama at BirminghamM. Ebald - University of IowaZ. Feng - University of IowaP. Wu - University of Alabama at BirminghamS. Park - University of Alabama at BirminghamX. Sun - University of Alabama at BirminghamZ. Yan - University of Alabama at BirminghamJ. Engelhardt - University of Alabama at BirminghamY. Tang - University of Alabama at Birmingham
- Resource Type
- Abstract
- Publication Details
- Journal of cystic fibrosis, Vol.25(Suppl 2), pp.117-117
- DOI
- 10.1016/j.jcf.2026.07.259
- ISSN
- 1569-1993
- Publisher
- Elsevier B.V
- Number of pages
- 1
- Language
- English
- Date published
- 09/2026
- Academic Unit
- Anatomy and Cell Biology
- Record Identifier
- 9985236349602771
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