Journal article
Increasing Epithelial Junction Permeability Enhances Gene Transfer to Airway Epithelia In Vivo
American journal of respiratory cell and molecular biology, Vol.22(2), pp.129-138
01/01/2000
DOI: 10.1165/ajrcmb.22.2.3938
PMID: 10657931
Abstract
Gene transfer to airway epithelia is the most direct approach for treating the progressive lung disease associated with cystic fibrosis. However, the transduction efficiency is poor when viral vectors are applied to the mucosal surface. We reported previously that gene transfer via the apical surface of human airway epithelia in vitro was improved by formulating vectors with ethyleneglycol-bis-(2-aminoethyl ether)- N,N,N',N'-tetraacetic acid (EGTA) in a hypotonic buffer. First, we investigated the mechanism for this enhancement. When 100-nm fluorescent beads were applied to the apical surface in the presence of EGTA, paracellular deposition of the particles was noted. Transmission electron microscopy verified that the epithelial junction complex was disrupted under these conditions. The Ca(2+) chelators EGTA, 1,2-bis (2-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA), and ethylenediaminetetraacetic acid all caused a rapid, reversible drop in transepithelial resistance and facilitated gene transfer with retrovirus or adenovirus in vitro. When Ca(2+) chelators were applied to rabbit tracheal epithelia or human nasal epithelia in vivo, the transepithelial voltage decreased, and amiloride sensitivity was lost, suggesting that epithelial junctions opened. Importantly, this novel formulation enhanced both retroviral- and adenoviral-mediated gene transfer to rabbit tracheal epithelia in vivo. This technique may have applications for vector or drug delivery to airway epithelia and other polarized cells.
Details
- Title: Subtitle
- Increasing Epithelial Junction Permeability Enhances Gene Transfer to Airway Epithelia In Vivo
- Creators
- Guoshun WangJoseph ZabnerCamille DeeringJan LaunspachJianqiang ShaoMordechai BodnerDoug J JollyBeverly L DavidsonPaul B Mccray
- Resource Type
- Journal article
- Publication Details
- American journal of respiratory cell and molecular biology, Vol.22(2), pp.129-138
- Publisher
- American Thoracic Society
- DOI
- 10.1165/ajrcmb.22.2.3938
- PMID
- 10657931
- ISSN
- 1044-1549
- eISSN
- 1535-4989
- Language
- English
- Date published
- 01/01/2000
- Academic Unit
- Pulmonary, Critical Care, and Occupational Medicine; Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine
- Record Identifier
- 9984093343002771
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