CFTR transgene expression in primary ΔF508 epithelial cell cultures from human nasal polyps following gene transfer with cationic phosphonolipids
Résumé
Cystic fibrosis (CF) is the most common autosomal lethal recessive disorder in the Caucasian population. The major cause of mortality is lung disease, owing to the failure of a functional protein from the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Today, even though the knowledge about the CFTR genomic is extensive, no efficient treatment has been developed yet. In this context, gene therapy represents a potential important advance on condition that it could develop efficient and safe transfection agents. Even though viral vectors have been used in most clinical trials owing to their high transfection efficiency, random integration and immunogenicity are still critical side effects. Consequently, all of these drawbacks brought forth the development of nonviral transfection systems. Although they engender few toxicity and immunogenicity problems, their low transfection efficiency is a hurdle that must be overcome. Over the past decade, we have developed an original family of monocationic lipids, cationic phosphonolipids, whose efficiency has been previously demonstrated both in vitro and in vivo. In this report, we observe that a new cationic phosphonolipid (KLN 30) can lead to the restoration of the CFTR protein following the ex vivo transfection of epithelial cells issuing from a ΔF508 homozygous patient. The transgene expression and the cytotoxicity correlate with the charge ratio of the lipoplex. A kinetic study was performed, and a luminescent signal was detected until 35 d after transfection. © 2004 Humana Press Inc. All rights of any nature whatsoever reserved.
Mots clés
transmembrane conductance regulator
unclassified drug
CFTR protein
human
liposome
phospholipid
phosphonolipids
recombinant protein
article
cell culture
controlled study
correlation analysis
cytotoxicity
epithelium cell
ex vivo study
gene transfer
genetic transfection
homozygote
human cell
human tissue
kinetics
luminescence
nose polyp
patient
signal detection
transgene
cytology
genetics
methodology
nose mucosa
pathology
Cations
Cultured
Cystic Fibrosis
Cystic Fibrosis Transmembrane Conductance Regulator
Epithelial Cells
Gene Expression
Gene Therapy
Gene Transfer Techniques
Humans
Liposomes
Nasal Mucosa
Nasal Polyps
Phospholipids
Recombinant Proteins
Transgenes
Biological membranes
Genetic engineering
Immunology
Population statistics
Pulmonary diseases
Autosomal disorder
Cystic fibrosis transmembrane conductance regulator (CFTR)
Monocationic lipids
Transfection system
Cells
1
2 dioleoyl 3 phosphophatidylethanolamine
2 dioleoyl 3 trimethylammoniopropane
cation
kln 30
lipid
lipoplex