Hepatocyte-targeted in vivo gene expression by intravenous injection of plasmid DNA complexed with synthetic multi-functional gene delivery system
Hepatocyte-targeted in vivo gene expression by intravenous injection of plasmid DNA complexed with synthetic multi-functional gene delivery system
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DOI：
10.1038/sj.gt.3301140
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发表时间：
2000-04-01
期刊：
影响因子：
5.1
通讯作者：
Hashida, M
中科院分区：
文献类型：
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作者：
Nishikawa, M;Yamauchi, M;Hashida, M
为了实现肝细胞靶向的体内基因表达，设计并合成了一种同时控制组织和细胞内DNA分布的载体。一种阳离子聚合物poly（l-鸟氨酸）（pOrn）首先用半乳糖修饰，然后用一个自生肽（mHA2）修饰，得到Gal-pOrn-mHA2。当将Gal-pOm-mHAP应用于asialal糖蛋白受体阳性细胞时，荧光素标记的DNA显示出弥漫性特征，表明载体从核内体和/或溶酶体释放DNA。然后检测静脉注射DNA复合物（每只小鼠10 μ g DNA）后的生物分布和基因表达。注射[P-32]DNA/Gal-pOrn-mHA2后，肝脏中约60%的放射性恢复，主要在实质细胞中。在注射DNA/Gal-pOrn-mHA2的小鼠肝脏中检测到大量转基因产物（荧光素酶）（81 ng/g组织），比注射DNA/DOTMA:Chol脂质体（50 μ g DNA）获得的荧光素酶多280倍。先前给予半乳糖化白蛋白可将基因表达降低至1/100，表明asialal糖蛋白受体介导的基因在肝实质细胞（即肝细胞）中转移。肝细胞中的荧光素酶活性占所有检查组织中总活性的95%以上。因此，通过静脉注射带有多功能基因载体的DNA复合物，实现了肝细胞靶向的体内基因表达。
To achieve hepatocyte-targeted in vivo gene expression, a carrier that controls both the tissue and intracellular distribution of DNA was designed and synthesized. A cationic polymer, poly(L-ornithine) (pOrn), was modified first with galactose, then with a fusigenic peptide (mHA2) to obtain Gal-pOrn-mHA2. When applied with Gal-pOm-mHAP to asialoglycoprotein receptor-positive cells, fluorescein-labeled DNA showed a diffuse profile, suggesting the release of DNA from endosomes and/or lysosomes by the carrier. Then the biodistribution and gene expression after intravenous injection of DNA complexes (10 mu g DNA per mouse) were examined. After injection of [P-32]DNA/Gal-pOrn-mHA2, about 60% of the radioactivity was recovered in the liver, mostly in parenchymal cells. A large amount (81 ng/g tissue) of transgene product (luciferase) was detected in the liver of mice injected with DNA/Gal-pOrn-mHA2, which was 280-fold greater than that obtained with DNA/DOTMA:Chol liposomes (50 mu g DNA). Prior administration of galactosylated albumin reduced the gene expression to 1/100, indicating the asialoglycoprotein receptor-mediated gene transfer in liver parenchymal cells, ie hepatocytes. The luciferase activity in hepatocytes contributed more than 95% of the total activity in all the tissues examined. Thus, hepatocyte-targeted in vivo gene expression was achieved by the intravenous injection of DNA complex with the multifunctional gene carrier.