In vivo gene delivery to the liver using novel galactosylated cationic liposomes
In vivo gene delivery to the liver using novel galactosylated cationic liposomes
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DOI：
10.1023/a:1007501122611
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发表时间：
2000-03-01
影响因子：
3.7
通讯作者：
Hashida, M
中科院分区：
文献类型：
--
作者：
Kawakami, S;Fumoto, S;Hashida, M
目的。本研究的目的是阐明含有胆甾烯-5-基氧基-N(4-((1-亚氨基-2-β-D-硫代半乳糖基乙基)氨基)丁基)甲酰胺(Gal-C4-Chol)的半乳糖基化脂质体的体内基因转移与脂质组成和电荷比的关系。方法。制备了含有N-[1-(2,3-二油酰氧基)丙基]-n,n,n-三甲基氯化铵(DOTMA)、Gal-C4-Chol和胆固醇(Chol)的半乳糖基化阳离子脂质体以及类似的脂质体。将与半乳糖基化脂质体制剂复合的质粒DNA注射到小鼠体内。 6小时后处死小鼠。对组织进行荧光素酶测定。结果。注射与 DOTMA/Chol/Gal-C4-Chol(1:0.5:0.5) 和 DOTMA/Gal-C4-Chol(1:1) 脂质体复合的质粒 DNA 后，肝脏中的基因表达显着升高。其效果比裸 DNA 和 DOTMA/Chol(1:1) 脂质体高一个数量级。预暴露于半乳糖基化牛血清白蛋白显着降低了肝基因表达。相比之下，含有3β[N-(N',N'-二甲基氨基乙烷)氨基甲酰基]胆固醇、Gal-C4-Chol和二油酰磷脂酰乙醇胺的半乳糖基化阳离子脂质体的基因表达低10倍。以DOTMA/Chol/Gal-CA-Chol(1:0.5:0.5)脂质体与质粒DNA(1.6-7.0)的电荷比而言，电荷比为2.3-3.1的复合物在肝脏中产生最大的基因表达。然而，较高的比率导致肺部表达增强。结论。通过优化脂质组成和电荷比，半乳糖基化脂质体/DNA 复合物可通过脱唾液酸糖蛋白受体介导的内吞作用在肝脏中实现卓越的体内基因转染。
Purpose. 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-thiogalaclosylethyl)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 3 beta[N-(N',N'-dimethylaminoethane)carbamoyl]cholesterol, Gal-C4-Chol and dioleoylphosphatidylethanolamine was 10 times lower. As Ear 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.