CFTR transgene expression in primary DeltaF508 epithelial cell cultures from human nasal polyps following gene transfer with cationic phosphonolipids.
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 F508 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.
Montier
Tristan
T
Université de Bretagne Occidentale, Institute de Synergie des Sciences et de la Santé and 2UNR CNRS 6521, Faculté des Sciences, Brest, France. Tristan.Montier@univ-brest.fr
Delépine
Pascal
P
Marianowski
Rémi
R
Le Ny
Karine
K
Le Bris
Morgane
M
Gillet
Danielle
D
Potard
Gaël
G
Mondine
Philippe
P
Frachon
Irène
I
Yaouanc
Jean-Jacques
JJ
Clément
Jean-Claude
JC
Des Abbayes
Harvé
H
Férec
Claude
C
D002412:Cations; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004847:Epithelial Cells / Q000166:cytology / Q000473:pathology; D015870:Gene Expression / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D008081:Liposomes; D009297:Nasal Mucosa / Q000166:cytology* / Q000473:pathology; D009298:Nasal Polyps / Q000235:genetics / Q000473:pathology; D010743:Phospholipids; D011994:Recombinant Proteins / Q000235:genetics; D019076:Transgenes / Q000235:genetics
C505032:CFTR protein, human; D002412:Cations; D008081:Liposomes; D010743:Phospholipids; D011994:Recombinant Proteins; C028299:phosphonolipids; D019005:Cystic Fibrosis Transmembrane Conductance Regulator