Enhanced hepatocyte-selective in vivo gene expression by stabilized galactosylated liposome/plasmid DNA complex using sodium chloride for complex formation
Enhanced hepatocyte-selective in vivo gene expression by stabilized galactosylated liposome/plasmid DNA complex using sodium chloride for complex formation
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DOI：
10.1016/j.ymthe.2004.07.015
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发表时间：
2004-10-01
影响因子：
12.4
通讯作者：
Hashida, M
中科院分区：
文献类型：
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作者：
Fumoto, S;Kawakami, S;Hashida, M
在这项研究中，我们证明了在阳离子脂质体/质粒DNA复合物（lipoplexes）的形成过程中，基本量的氯化钠（NaCl）的存在下，稳定的lipoplexes根据表面电荷调节（SCR）理论。荧光共振能量转移分析表明，在SCR lipoplex（5和10 mM NaCl溶液的lipoplex）的阳离子脂质体增加融合。此外，SCR lipoplexes的聚集后，暴露于生理盐水（150 mM NaCl）作为生理条件的模型显着延迟。门静脉内给药后，半乳糖化的SCR lipoplex（5和10 mM NaCl溶液的lipoplex）的肝转染活性约为10- 20倍，高于小鼠中的半乳糖化的常规lipoplex。半乳糖化SCR lipoplexes在肝细胞中的转染活性显著高于常规lipoplexes，并且预先暴露于竞争性脱唾液酸糖蛋白受体阻断剂显著降低了肝基因表达，表明肝细胞负责半乳糖化SCR lipoplexes的高肝转基因表达。原位和体内的药代动力学研究表明，半乳糖基化的SCR脂质复合物具有更高的组织结合亲和力和更广泛的肝内分布。此外，在HepG 2细胞中观察到半乳糖基化的SCR lipoplexes的增强的转染活性，并且共聚焦显微镜图像的调查显示，在细胞中的质粒DNA的释放显著加速。这些特征部分解释了半乳糖基化的SCR脂质复合物增强体内转染功效的机制。因此，在这项研究中的信息将是有价值的配体修饰的lipoplexes在体内应用的未来使用，设计和开发。
In this study, we demonstrated that the presence of an essential amount of sodium chloride (NaCl) during the formation of cationic liposome/plasmid DNA complexes (lipoplexes) stabilizes the lipoplexes according to the surface charge regulation (SCR) theory. Fluorescence resonance energy transfer analysis revealed that cationic liposomes in an SCR lipoplex (5 and 10 mM NaCl solution in lipoplex) increased fusion. Also, aggregation of SCR lipoplexes was significantly delayed after exposure to saline (150 mM NaCl) as a model of physiological conditions. After intraportal administration, the hepatic transfection activity of galactosylated SCR lipoplexes (5 and 10 mM NaCl solution in lipoplex) was approximately 10- to 20-fold higher than that of galactosylated conventional lipoplexes in mice. The transfection activity in hepatocytes of galactosylated SCR lipoplexes was significantly higher than that of conventional lipoplexes, and preexposure to competitive asialoglycoprotein-receptor blocker significantly reduced the hepatic gene expression, suggesting that hepatocytes are responsible for high hepatic transgene expression of the galactosylated SCR lipoplexes. Pharmacokinetic studies both in situ and in vivo demonstrated a higher tissue binding affinity and a greater expanse of intrahepatic distribution by galactosylated SCR lipoplexes. Moreover, enhanced transfection activity of galactosylated SCR lipoplexes was observed in HepG2 cells, and investigation of confocal microscopic images showed that the release of plasmid DNA in the cell was markedly accelerated. These characteristics partly explain the mechanism of enhanced in vivo transfection efficacy by galactosylated SCR lipoplexes. Hence, information in this study will be valuable for the future use, design, and development of ligand-modified lipoplexes for in vivo applications.