Targeting transgene expression for cystic fibrosis gene therapy

Citation
Dr. Koehler et al., Targeting transgene expression for cystic fibrosis gene therapy, MOL THER, 4(1), 2001, pp. 58-65
Citations number
49
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Molecular Biology & Genetics
Journal title
MOLECULAR THERAPY
ISSN journal
1525-0016 → ACNP
Volume
4
Issue
1
Year of publication
2001
Pages
58 - 65
Database
ISI
SICI code
1525-0016(200107)4:1<58:TTEFCF>2.0.ZU;2-J
Abstract
We have developed an expression cassette for cystic fibrosis (CF) gene ther apy using control elements from the human cytokeratin 18 gene (KRT18, also known as K18). KRT18 is naturally expressed in a spatial pattern similar to that of CFTR, the gene mutated in CF. We delivered a KRT18-driven lad. pla smid complexed with cationic liposomes intravenously to mice and examined e xpression in various tissues. We found expression in nasal and bronchial ep ithelium, airway submucosal glands, gall bladder, and kidneys. Expression w as low in pancreas and gut, and absent from liver and alveolar lung. This i s consistent with the expression pattern reported for a K18lacZ transgenic moose. Following delivery of a cytomegalovirus (CMV) major immediate-early promoter/enhancer-driven lacZ plasmid, we found expression in bronchi, subm ucosal glands, alveolar cells, liver, and kidney. We did not detect express ion in nose, pancreas, gall bladder, or gut. Using fluorescently labeled pl asmid delivered by means of liposomes, we identified the liver, alveolar lu ng, and kidneys as the major plasmid deposition sites. Our data demonstrate that a KRT18-driven expression vector delivered systemically can target ge ne expression to CF-affected tissues, despite an uneven distribution of pla smid DNA. A KRT18-based vector may be a useful alternative to viral promote r-based vectors in clinical gene therapy trials to treat CF.