Journal article
Adenovirus 5-fiber 35 chimeric vector mediates efficient apical correction of the cystic fibrosis transmembrane conductance regulator defect in cystic fibrosis primary airway epithelia
Human gene therapy, Vol.21(3), pp.251-269
03/2010
DOI: 10.1089/hum.2009.056
PMID: 19788389
Abstract
In vivo gene transfer to the human respiratory tract by adenovirus serotype 5 (Ad5) vectors has revealed their limitations related to inefficient gene transfer, host antiviral response, and innate adenoviral toxicity. In the present work, we compared the cytotoxicity and efficiency of Ad5 and a chimeric Ad5F35 vector with respect to CFTR gene transfer to cystic fibrosis (CF) and non-CF human airway epithelial cells. We found that high doses of Ad5 vector had an adverse effect on the function of exogenous and endogenous CFTR. Results obtained with Ad5 capsid mutants suggested that the RGD motifs on the penton base capsomers were responsible for the negative effect on CFTR function. This negative interference did not result from a lower level of biosynthesis and/or altered cellular trafficking of the CFTR protein, but rather from an indirect mechanism of functional blockage of CFTR, related to the RGD integrin-mediated endocytic pathway of Ad5. No negative interference with CFTR was observed for Ad5F35, an Ad5-based vector pseudotyped with fibers from Ad35, a serotype that uses another cell entry pathway. In vitro, Ad5F35 vector expressing the GFP-tagged CFTR (Ad5F35-GFP-CFTR) showed a 30-fold higher efficiency of transduction and chloride channel correction in CFTR-deficient cells, compared with Ad5GFP-CFTR. Ex vivo, Ad5F35-GFP-CFTR had the capacity to transduce efficiently reconstituted airway epithelia from patients with CF (CF-HAE) via the apical surface, restored chloride channel function at relatively low vector doses, and showed relatively stable expression of GFP-CFTR for several weeks.
Details
- Title: Subtitle
- Adenovirus 5-fiber 35 chimeric vector mediates efficient apical correction of the cystic fibrosis transmembrane conductance regulator defect in cystic fibrosis primary airway epithelia
- Creators
- Ophélia Granio - Virologie et Pathologie HumaineJoseph Zabner - Department of MedicineKatherine J D Ashbourne Excoffon - Department of MedicinePetra Henning - Department of Microbiology and ImmunologyPatricia Melin - Institut de physiologie et biologie cellulairesCaroline Norez - Institut de physiologie et biologie cellulairesGaëlle Gonzalez - Virologie et Pathologie HumainePhilip H Karp - Department of MedicineMaria K Magnusson - Department of Microbiology and ImmunologyNagy Habib - Department of Surgical Oncology and Technology [London]Leif Lindholm - Department of Microbiology and ImmunologyFrédéric Becq - Institut de physiologie et biologie cellulairesPierre Boulanger - Virologie et Pathologie HumaineSaw-See Hong - Virologie et Pathologie Humaine
- Resource Type
- Journal article
- Publication Details
- Human gene therapy, Vol.21(3), pp.251-269
- Publisher
- Mary Ann Liebert
- DOI
- 10.1089/hum.2009.056
- PMID
- 19788389
- ISSN
- 1043-0342
- eISSN
- 1557-7422
- Language
- English
- Date published
- 03/2010
- Academic Unit
- Pulmonary, Critical Care, and Occupational Medicine; Internal Medicine
- Record Identifier
- 9984094367202771
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