Journal article
Postentry Processing of Recombinant Adeno-Associated Virus Type 1 and Transduction of the Ferret Lung Are Altered by a Factor in Airway Secretions
Human gene therapy, Vol.24(9), pp.786-796
09/01/2013
DOI: 10.1089/hum.2013.137
PMCID: PMC3768237
PMID: 23948055
Abstract
We recently created a cystic fibrosis ferret model that acquires neonatal lung infection. To develop lung gene therapies for this model, we evaluated recombinant adeno-associated virus (rAAV)-mediated gene transfer to the neonatal ferret lung. Unlike
in vitro
ferret airway epithelial (FAE) cells,
in vivo
infection of the ferret lung with rAAV1 required proteasome inhibitors to achieve efficient airway transduction. We hypothesized that differences in transduction between these two systems were because of an
in vivo
secreted factor that alter the transduction biology of rAAV1. Indeed, treatment of rAAV1 with ferret airway secretory fluid (ASF) strongly inhibited rAAV1, but not rAAV2, transduction of primary FAE and HeLa cells. Properties of the ASF inhibitory factor included a strong affinity for the AAV1 capsid, heat-stability, negative charge, and sensitivity to endoproteinase Glu-C. ASF-treated rAAV1 dramatically inhibited apical transduction of FAE ALI cultures (512-fold), while only reducing viral entry by 55-fold, suggesting that postentry processing of virus was influenced by the inhibitor factor. Proteasome inhibitors rescued transduction in the presence of ASF (∼1600-fold) without effecting virus internalization, while proteasome inhibitors only enhanced transduction 45-fold in the absence of ASF. These findings demonstrate that a factor in lung secretions can influence intracellular processing of rAAV1 in a proteasome-dependent fashion.
Yan and colleagues identify a protease-sensitive factor in ferret airway secretions that profoundly reduces transduction by AAV1. The unidentified factor inhibits both vector entry and postentry processing in polarized ferret airway epithelial (FAE) cell cultures. Transduction can be rescued both
in vitro
and
in vivo
by a proteasome inhibitor, suggesting this novel factor interacts with the ubiquitin–proteasome system during intracellular virus processing.
Details
- Title: Subtitle
- Postentry Processing of Recombinant Adeno-Associated Virus Type 1 and Transduction of the Ferret Lung Are Altered by a Factor in Airway Secretions
- Creators
- Ziying Yan - 2Center for Gene Therapy, University of Iowa School of Medicine, Iowa City, IA 52242-1009Xingshen Sun - 1Department of Anatomy and Cell Biology, University of Iowa School of Medicine, Iowa City, IA 52242-1009Idil A Evans - 1Department of Anatomy and Cell Biology, University of Iowa School of Medicine, Iowa City, IA 52242-1009Scott R Tyler - 1Department of Anatomy and Cell Biology, University of Iowa School of Medicine, Iowa City, IA 52242-1009Yi Song - 1Department of Anatomy and Cell Biology, University of Iowa School of Medicine, Iowa City, IA 52242-1009Xiaoming Liu - 2Center for Gene Therapy, University of Iowa School of Medicine, Iowa City, IA 52242-1009Hongshu Sui - 1Department of Anatomy and Cell Biology, University of Iowa School of Medicine, Iowa City, IA 52242-1009John F Engelhardt - 3Department of Internal Medicine, University of Iowa School of Medicine, Iowa City, IA 52242-1009
- Resource Type
- Journal article
- Publication Details
- Human gene therapy, Vol.24(9), pp.786-796
- DOI
- 10.1089/hum.2013.137
- PMID
- 23948055
- PMCID
- PMC3768237
- NLM abbreviation
- Hum Gene Ther
- ISSN
- 1043-0342
- eISSN
- 1557-7422
- Publisher
- Mary Ann Liebert, Inc
- Language
- English
- Date published
- 09/01/2013
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984025345302771
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