Functional evidence of CFTR gene transfer in nasal epithelium of cystic fibrosis mice in vivo following luminal application of DNA complexes targeted to the serpin-enzyme complex receptor.
Molecular conjugates that target the serpin-enzyme complex receptor transfer the cDNA encoding human cystic fibrosis transmembrane conductance regulator (CFTR) to the nasal epithelium of cystic fibrosis mutant mice. These complexes effect partial correction of the chloride transport defect as assessed by in vivo nasal potential difference measurements, produce immunohistochemical staining for CFTR, and restore expression of nitric oxide synthase-2 (NOS-2), which is downregulated in the epithelium of mice and humans with cystic fibrosis. Complexes that lack the receptor ligands were ineffective, so receptor access was essential. Mice treated with receptor-targeted lacZ showed beta-galactosidase expression in epithelial cells and submucosal glands, but no electrophysiologic correction or NOS-2 expression, so simply accessing the serpin-enzyme complex receptor was not sufficient to produce the observed electrophysiologic or immunohistochemical changes. Correction of the cAMP-stimulated chloride transport was dose related at days 7 and 12 after complex administration, but, for most animals, nasal potential difference had returned to baseline by day 18. Molecular conjugates targeting the serpin-enzyme complex receptor, used to compact plasmid DNA, hold promise for gene therapy of cystic fibrosis.
Ziady
Assem-Galal
AG
Department of Pediatrics at Rainbow Babies and Childrens Hospital, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA.
Kelley
Thomas J
TJ
Milliken
Erin
E
Ferkol
Thomas
T
Davis
Pamela B
PB
D000818:Animals; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000628:therapy; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004594:Electrophysiology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D018181:Mice, Inbred CFTR; D009297:Nasal Mucosa / Q000201:enzymology / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
C505032:CFTR protein, human; D011956:Receptors, Cell Surface; C064184:serpin-enzyme complex receptor; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D001616:beta-Galactosidase