In vitro and in vivo gene transfer to pulmonary cells mediated by cationic liposomes.
Cationic liposomes have been proposed as alternative to adenovirus in the treatment of cystic fibrosis lung disease. Therefore, we have investigated the efficiency of two lipid mixtures in mediating gene transfer in in vitro and in vivo models. The cationic lipid DOTMA (N-(1-(2,3(dioleyloxy)propyl)-n,n,n-trimethylammoniumchloride++ +) and DOGS (dioctadecylamidoglycylspermine) were used in combination with the neutral lipid DOPE (dioleoylphosphatidylethanolamine). The relative transfection efficiencies of the two cationic liposomes were tested using the bacterial beta-galactosidase (lacZ) and the firefly luciferase genes. Gene expression was detected in both cell limes and primary culture of rhesus monkey airway epithelium after transfection with plasmid DNA complexed with DOGS/DOPE or DOTMA/DOPE. Transfection efficiency of both types of lipids was higher in the mouse fibroblast 3T3 cell line as compared to human carcinoma A549 cells and primary epithelial cultures. Administration of DNA-liposome complexes via intratracheal instillation resulted in expression of the lacZ and luciferase marker gene in the mouse airways. In vivo transfection mediated by both types of liposomes were proven to be far less efficient than adenovirus treatment.
Fortunati
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Department of Pediatrics, CRIBI Biotechnology Center, Padova, Italy. fortunati@ch.fgg.eur.nl
Bout
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Zanta
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Valerio
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Scarpa
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D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001980:Bronchi / Q000378:metabolism; D002412:Cations; D002460:Cell Line; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000628:therapy; D004274:DNA, Recombinant / Q000008:administration & dosage* / Q000235:genetics; D004337:Drug Carriers; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D005998:Glycine / Q000031:analogs & derivatives; D006801:Humans; D007322:Instillation, Drug; D008081:Liposomes / Q000008:administration & dosage; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008168:Lung / Q000166:cytology*; D008253:Macaca mulatta; D051379:Mice; D008807:Mice, Inbred BALB C; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013096:Spermine / Q000031:analogs & derivatives; D014132:Trachea / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D002412:Cations; D004274:DNA, Recombinant; D004337:Drug Carriers; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D011993:Recombinant Fusion Proteins; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium; C061134:dioctadecylamidoglycylspermine; D013096:Spermine; C020888:1,2-dielaidoylphosphatidylethanolamine; D008156:Luciferases; D001616:beta-Galactosidase; D005998:Glycine