Stabilized plasmid-lipid particles for systemic gene therapy
Stabilized plasmid-lipid particles for systemic gene therapy
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DOI：
10.1038/sj.gt.3301308
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发表时间：
2000-11-01
期刊：
影响因子：
5.1
通讯作者：
Cullis, PR
中科院分区：
文献类型：
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作者：
Tam, P;Monck, M;Cullis, PR
研究了“稳定的质粒-脂质颗粒”（SPLP）的结构及其作为系统性基因治疗载体的性质。我们表明，SPLP可以可视化采用冷冻电子显微镜是均匀的颗粒直径72 +/- 5 nm的脂质双层包围质粒DNA的核心组成。还显示SPLP在静脉内（i. v.）在小鼠肿瘤模型中注射，导致高达3%的总注射剂量的累积和在远端（后胁腹）肿瘤部位的伴随报告基因表达。相比之下，静脉注射裸质粒DNA或质粒DNA-阳离子脂质体复合物不会导致显著的质粒递送至肿瘤部位或在该部位的基因表达。此外，通过监测血清酶水平测定，表明相当于每只小鼠175 μ g质粒的高剂量SPLP是无毒的，而静脉内注射复合物在每只小鼠20 μ g质粒以上的剂量水平产生显著毒性。它的结论是，SPLP表现出与潜在的效用作为一个无毒的系统性基因治疗载体的属性一致。
The structure of 'stabilized plasmid-lipid particles' (SPLP) and their properties as systemic gene therapy vectors has been investigated. We show that SPLP can be visualized employing cryo-electron microscopy to be homogeneous particles of diameter 72 +/- 5 nm consisting of a lipid bilayer surrounding a core of plasmid DNA. It is also shown that SPLP exhibit long circulation lifetimes (circulation half-life >6 h) following intravenous (i.v.) injection in a murine tumor model resulting in accumulation of up to 3% of the total injected dose and concomitant reporter gene expression at a distal (hind flank) tumor site. In contrast, i.v. injection of naked plasmid DNA or plasmid DNA-cationic liposome complexes did not result in significant plasmid delivery to the tumor site or gene expression at that site. Furthermore, it is shown that high doses of SPLP corresponding to 175 mug plasmid per mouse are nontoxic as assayed by monitoring serum enzyme levels, whereas i.v. injection of complexes give rise to significant toxicity at dose levels above 20 mug plasmid per mouse. It is concluded that SPLP exhibit properties consistent with potential utility as a nontoxic systemic gene therapy vector.