title: Increased contact time improves adenovirus-mediated CFTR gene transfer to nasal epithelium of CF mice.
abstract: Multiple dosing with recombinant adenoviral vectors containing the cystic fibrosis transmembrane conductance regulator (CFTR) cDNA to the nasal mucosa of cystic fibrosis (CF) transgenic mice reportedly results in only partial correction of the CF defect in chloride (Cl-) secretion without normalizing sodium (Na+) hyperabsorption, perhaps indicating inefficient gene transfer into the nasal airway epithelium in vivo. In this study, we have examined whether optimizing vector administration such as contact time could improve gene transfer efficiency. Changes in basal nasal potential difference (PD), and in PD (delta PD) following addition of amiloride and subsequent removal of Cl- from the luminal perfusate were assayed. As reported previously, the basal nasal PD was significantly more negative in CF mice (-24.9 +/- 2.1 mV) than in normal mice (-6.3 +/- 1.2 mV). Normal mouse nasal mucosa exhibited a large hyperpolarization in response to low Cl- substitution (delta PD of 8.5 +/- 1.9 mV), whereas the nasal mucosa of the CF mouse depolarized in response to this treatment. No correction of either the Cl- or Na+ transport defects were observed when 5 x 10(9) IU of Ad2/CFTR-5 were administered to the nasal passage of CF mice over a period of 5-20 min. However, when CF mice were perfused over a period of 60 min with the same dose of vector, a significant response (delta PD of 5.9 +/- 1.1 mV) to low Cl- substitution was detected 2 days later. In these mice, the basal nasal PD (-10.5 +/- 1.4 mV) and the response to amiloride were also reduced, indicating a partial correction of the Na+ transport defect. Expression of functional CFTR activity was transient with no measurable delta PD signals observed by day 7 post-treatment. These results suggest that prolonging the contact between an adenoviral vector and the respiratory epithelium enhances the efficiency of gene transfer and can result in improved correction of the CF Na+ and Cl- ion transport defects. Therefore, strategies that improve internalization of viral vectors and that prolong their contact time with target cells may result in the improved clinical efficacy of such vectors.
authors.affiliation: Genzyme Corporation, Framingham, MA 01701-9322, USA.
authors.forename: C
authors.initials: C
authors.lastname: Jiang
authors.forename: G Y
authors.initials: GY
authors.lastname: Akita
authors.forename: W H
authors.initials: WH
authors.lastname: Colledge
authors.forename: R A
authors.initials: RA
authors.lastname: Ratcliff
authors.forename: M J
authors.initials: MJ
authors.lastname: Evans
authors.forename: K M
authors.initials: KM
authors.lastname: Hehir
authors.forename: J A
authors.initials: JA
authors.lastname: St George
authors.forename: S C
authors.initials: SC
authors.lastname: Wadsworth
authors.forename: S H
authors.initials: SH
authors.lastname: Cheng
mesh_terms: D000256:Adenoviridae; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D018118:Chloride Channels / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D018076:DNA, Complementary / Q000008:administration & dosage; D004232:Diuretics / Q000494:pharmacology; D004848:Epithelium / Q000378:metabolism; D018014:Gene Transfer Techniques; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009297:Nasal Mucosa / Q000166:cytology*; D012964:Sodium / Q000378:metabolism; D013997:Time Factors
chemical_list: D018118:Chloride Channels; D002712:Chlorides; D018076:DNA, Complementary; D004232:Diuretics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000584:Amiloride; D012964:Sodium