title: A novel cationic lipid greatly enhances plasmid DNA delivery and expression in mouse lung.
abstract: Effective gene therapy for lung tissue requires the use of efficient vehicles to deliver the gene of interest into lung cells. When plasmid DNA encoding chloramphenicol acetyltransferase (CAT) was administered intranasally to BALB/c mice without carrier lipids, CAT activity was detected in mouse lung extracts. Plasmid DNA delivered with optimally formulated commercially available transfection reagents expressed up to 10-fold more CAT activity in lung than observed with naked DNA alone. Liposome formulations consisting of (+/-)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis (dodecyloxy)-1-propanaminium bromide (GAP-DLRIE) plus the neutral colipid dioleoylphosphatidylethanolamine (DOPE) enhanced CAT expression by more than 100-fold relative to plasmid DNA alone. A single administration of GAP-DLRIE liposome-CAT DNA complexes to mouse lung elicited peak expression at days 1-4 posttransfection, followed by a gradual return to baseline by day 21 postadministration. Readministration of GAP-DLRIE liposome CAT complexes at day 21 led to another transient peak of reporter gene expression. Histological examination of lungs treated with GAP-DLRIE complexed beta-galactosidase DNA revealed that alveolar epithelial cells were the primary locus of expression and that up to 1% of all alveoli contained epithelial cells expressing the transgene.
authors.affiliation: Vical Inc., San Diego, CA 92121, USA.
authors.forename: C J
authors.initials: CJ
authors.lastname: Wheeler
authors.forename: P L
authors.initials: PL
authors.lastname: Felgner
authors.forename: Y J
authors.initials: YJ
authors.lastname: Tsai
authors.forename: J
authors.initials: J
authors.lastname: Marshall
authors.forename: L
authors.initials: L
authors.lastname: Sukhu
authors.forename: S G
authors.initials: SG
authors.lastname: Doh
authors.forename: J
authors.initials: J
authors.lastname: Hartikka
authors.forename: J
authors.initials: J
authors.lastname: Nietupski
authors.forename: M
authors.initials: M
authors.lastname: Manthorpe
authors.forename: M
authors.initials: M
authors.lastname: Nichols
authors.forename: M
authors.initials: M
authors.lastname: Plewe
authors.forename: X
authors.initials: X
authors.lastname: Liang
authors.forename: J
authors.initials: J
authors.lastname: Norman
authors.forename: A
authors.initials: A
authors.lastname: Smith
authors.forename: S H
authors.initials: SH
authors.lastname: Cheng
mesh_terms: D000281:Administration, Intranasal; D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D004337:Drug Carriers; D004848:Epithelium; D004987:Ethers / Q000138:chemical synthesis*; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D006651:Histocytochemistry; D006801:Humans; D008081:Liposomes; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D010714:Phosphatidylethanolamines; D010957:Plasmids / Q000008:administration & dosage*; D000644:Quaternary Ammonium Compounds / Q000138:chemical synthesis*; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D004337:Drug Carriers; D004987:Ethers; D008081:Liposomes; C101963:N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(dodecyloxy)-1-propanaminium; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C020888:1,2-dielaidoylphosphatidylethanolamine; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase