title: Novel cationic liposome formulation for the delivery of an oligonucleotide decoy to NF-kappaB into activated macrophages.
abstract: Nuclear factor-kappaB (NF-kappaB) is involved in several pathological processes, such as inflammation. Pro-inflammatory genes expression can be down-regulated by using an oligonucleotide (ODN) decoy to NF-kappaB. Cationic liposomes are largely used to improve ODN uptake into cells, although a higher transfection efficiency and a lower toxicity are required to use them in therapy. In this work, we investigated the potential of a novel liposome formulation, based on the recently synthesised cationic lipid (2,3-didodecyloxypropyl) (2-hydroxyethyl) dimethylammonium bromide (DE), as the delivery system for a double stranded ODN decoy to NF-kappaB. Liposomes composed of DE or DE mixed with 1,2-dioleyl-sn-glycero-3-phosphoethanolamine or cholesterol as helper lipids were complexed with ODN at different +/- charge ratios. In vitro uptake and the effect of ODN, naked or complexed with DE-containing liposomes, were evaluated in lipopolysaccharide-stimulated RAW 264.7 macrophages. The use of helper lipids increased liposome physical stability up to 1 year at 4 degrees C. ODN complexed with DE/cholesterol liposomes, at the +/- charge ratio of 8, showed a limited cytotoxicity and the highest inhibition of nitrite production, inducible nitric oxide synthase protein expression and NF-kappaB/DNA binding activity. Confocal microscopy confirmed a high ODN cell uptake obtained with DE/cholesterol liposomes at the highest +/- charge ratio.
authors.affiliation: Dipartimento di Chimica Farmaceutica e Tossicologica, Università degli Studi di Napoli Federico II, Naples, Italy. gderosa@unina.it
authors.forename: Giuseppe
authors.initials: G
authors.lastname: De Rosa
authors.forename: Daniela
authors.initials: D
authors.lastname: De Stefano
authors.forename: Valeria
authors.initials: V
authors.lastname: Laguardia
authors.forename: Silvia
authors.initials: S
authors.lastname: Arpicco
authors.forename: Vittorio
authors.initials: V
authors.lastname: Simeon
authors.forename: Rosa
authors.initials: R
authors.lastname: Carnuccio
authors.forename: Elias
authors.initials: E
authors.lastname: Fattal
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002412:Cations; D002460:Cell Line; D002470:Cell Survival; D002784:Cholesterol / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008081:Liposomes; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D009573:Nitrites / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism*; D010316:Particle Size; D010714:Phosphatidylethanolamines / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D014162:Transfection
chemical_list: D002412:Cations; D008055:Lipids; D008070:Lipopolysaccharides; D008081:Liposomes; D016328:NF-kappa B; D009573:Nitrites; D009841:Oligonucleotides; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C020888:1,2-dielaidoylphosphatidylethanolamine; D002784:Cholesterol