title: In vivo gene delivery to the liver using novel galactosylated cationic liposomes.
abstract: OBJECTIVE
  The purpose of this study is to elucidate the in vivo gene transfer for galactosylated liposomes containing cholesten-5-yloxy-N-(4-((1-imino-2-beta-D-thiogalactosyle thyl)amino)butyl)formamide(Gal-C4-Chol) in relation to lipid composition and charge ratio.
  
  
  METHODS
  Galactosylated cationic liposomes containing N-[1-(2,3-dioleyloxy)propyl]-n,n,n-trimethylammonium chloride(DOTMA), Gal-C4-Chol and cholesterol(Chol), and similar liposomes were prepared. Plasmid DNA complexed with a galactosylated liposome preparation was injected intraportally into mice. The mice were sacrificed after 6 hours. The tissues were subjected to luciferase assay.
  
  
  RESULTS
  A markedly higher gene expression in the liver following injection of plasmid DNA that has been complexed with DOTMA/ Chol/Gal-C4-Chol(1:0.5:0.5) and DOTMA/Gal-C4-Chol(1:1) liposomes was observed. The effect was one order of magnitude higher than naked DNA and DOTMA/Chol(1:1) liposomes. Pre-exposing with galactosylated bovine serum albumin significantly reduced the hepatic gene expression. By comparison, the gene expression for galactosylated cationic liposomes containing 3beta[N-(N',N'-dimethylaminoethane)carbamoyl]cholesterol, Gal-C4-Chol and dioleoylphosphatidylethanolamine was 10 times lower. As far as the charge ratio of DOTMA/ Chol/Gal-CA-Chol(1:0.5:0.5) liposomes to plasmid DNA(1.6-7.0) was concerned, complexes with charge ratios of 2.3-3.1 produced maximal gene expression in the liver. Whereas, higher ratios resulted in enhanced expression in the lung.
  
  
  CONCLUSIONS
  By optimizing lipid composition and charge ratio, galactosylated liposome/DNA complexes allow superior in vivo gene transfection in the liver via asialoglycoprotein receptor-mediated endocytosis.
authors.affiliation: Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Kawakami
authors.forename: S
authors.initials: S
authors.lastname: Fumoto
authors.forename: M
authors.initials: M
authors.lastname: Nishikawa
authors.forename: F
authors.initials: F
authors.lastname: Yamashita
authors.forename: M
authors.initials: M
authors.lastname: Hashida
mesh_terms: D000818:Animals; D037263:Asialoglycoprotein Receptor; D002412:Cations / Q000378:metabolism; D002478:Cells, Cultured; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000493:pharmacokinetics*; D005690:Galactose / Q000031:analogs & derivatives* / Q000378:metabolism*; D015870:Gene Expression / Q000502:physiology; D018014:Gene Transfer Techniques; D008081:Liposomes / Q000493:pharmacokinetics*; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008813:Mice, Inbred ICR; D010316:Particle Size; D010713:Phosphatidylcholines / Q000493:pharmacokinetics; D010957:Plasmids; D000644:Quaternary Ammonium Compounds / Q000493:pharmacokinetics; D011956:Receptors, Cell Surface / Q000378:metabolism; D014162:Transfection
chemical_list: D037263:Asialoglycoprotein Receptor; D002412:Cations; D008081:Liposomes; D010713:Phosphatidylcholines; D000644:Quaternary Ammonium Compounds; D011956:Receptors, Cell Surface; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium; D002784:Cholesterol; D005690:Galactose