Modulation of a membrane lipid lamellar-nonlamellar phase transition by cationic lipids: a measure for transfection efficiency.
Modulation of a membrane lipid lamellar-nonlamellar phase transition by cationic lipids: a measure for transfection efficiency.
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阳离子脂质对膜脂层状-非层状相变的调节：转染效率的衡量标准。
DOI：
10.1016/j.bbamem.2008.07.022
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
MacDonald,RobertC
中科院分区：
文献类型：
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作者：
Tenchov,BorisG;Wang,Li;Koynova,Rumiana;MacDonald,RobertC
合成阳离子脂质可以作为DNA的载体，被认为是最有前途的非病毒基因载体。本文合成了6种具有不同疏水基团的新型磷脂酰胆碱（PC）阳离子衍生物，并测定了其转染人脐动脉内皮细胞（huec）的效率。三种具有相对较短的肉豆蔻油酰基或肉豆蔻油酰基14:1链的化合物表现出非常高的活性，比参考阳离子衍生物二酰基乙基pc （EDOPC）高出约10倍。值得注意的是，在转染检测中，14:1碳氢链的阳离子脂质尚未作为DNA载体进行过测试。其他三种脂质含有油酰18:1和更长的链，表现出中等至弱的转染活性。转染效率与阳离子脂质对双棕榈油酰磷脂酰乙醇胺分散体（DPoPE）层状到倒六方（Lα→HII）相转化的影响密切相关。对二元DPoPE/阳离子脂质混合物的x射线衍射结果表明，优良的转染剂消除了Lα→hii相的直接转变，促进了Lα与hii相之间的倒立方相的形成。相比之下，中等和弱转染剂保持了直接的Lα→hii转变，但比纯DPoPE转移到更高的温度，并在后期诱导立方相形成。在目前脂质膜融合模型的基础上，高效试剂对立方相的促进可能表明它们的高转染活性是由于脂质体与细胞膜的融合增强。阳离子脂质转染效率与其调节膜脂的非层状相形成的方式之间建立了明显且表达良好的相关性，这可以作为评估脂质载体质量和合理设计新型和优质核苷酸递送剂的标准。
Synthetic cationic lipids can be used as DNA carriers and are regarded to be the most promising non-viral gene carriers. For this investigation, six novel phosphatidylcholine (PC) cationic derivatives with various hydrophobic moieties were synthesized and their transfection efficiencies for human umbilical artery endothelial cells (HUAEC) were determined. Three compounds with relatively short, myristoleoyl or myristelaidoyl 14:1 chains exhibited very high activity, exceeding by ∼10 times that of the reference cationic derivative dioleoyl ethylPC (EDOPC). Noteworthy, cationic lipids with 14:1 hydrocarbon chains have not been tested as DNA carriers in transfection assays previously. The other three lipids, which contained oleoyl 18:1 and longer chains, exhibited moderate to weak transfection activity. Transfection efficiency was found to correlate strongly with the effect of the cationic lipids on the lamellar-to-inverted hexagonal, Lα→HII, phase conversion in dipalmitoleoyl phosphatidylethanolamine dispersions (DPoPE). X-ray diffraction on binary DPoPE/cationic lipid mixtures showed that the superior transfection agents eliminated the direct Lα→HIIphase transition and promoted formation of an inverted cubic phase between the Lαand HIIphases. In contrast, moderate and weak transfection agents retained the direct Lα→HIItransition but shifted to higher temperatures than that of pure DPoPE, and induced cubic phase formation at a later stage. On the basis of current models of lipid membrane fusion, promotion of a cubic phase by the high-efficiency agents may be considered as an indication that their high transfection activity results from enhanced lipoplex fusion with cellular membranes. The distinct, well-expressed correlation established between transfection efficiency of a cationic lipid and the way it modulates nonlamellar phase formation of a membrane lipid could be useful as a criterion to assess the quality of lipid carriers and for rational design of new and superior nucleotide delivery agents.