Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cells.
Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cells.
复制标题
阳离子脂质包被的 PEI/DNA 复合物可提高基因递送至间充质干细胞的效率并降低细胞毒性
DOI：
10.2147/ijn.s33923
复制
发表时间：
2012
影响因子：
8
通讯作者：
Gu Z
中科院分区：
文献类型：
--
作者：
Song H;Wang G;He B;Li L;Li C;Lai Y;Xu X;Gu Z
背景 无需血清剥夺的有效基因转染是成功的基于干细胞的基因治疗的先决条件。聚乙烯亚胺（PEI）是一种有效的非病毒基因载体，但其应用受到血清敏感性和严重细胞毒性的阻碍。方法为了解决这个问题，通过用三种抗血清阳离子脂质（即赖氨酰化、组氨酰化和精氨酰化胆固醇）涂覆PEI/DNA复合物，开发了一个新的脂多聚复合物家族。研究了脂多聚复合物的物理性质、转染效率、细胞摄取、亚细胞分布和细胞毒性。结果在血清存在下，由赖氨酰化或组氨酰化胆固醇组成的外层显着提高了PEI/DNA比为2的低聚合复合物的转染效率。由此产生的赖氨酰化和组氨酰化胆固醇脂多聚复合物甚至比性能最佳的PEI/DNA比率为10的多聚复合物更有效。细胞摄取和亚细胞分布研究的结果表明，它们更高的转染效率可能是由于加速了DNA核定位。通过递送治疗基因 hVEGF165 也证实了脂多聚复合物相对于性能最佳的多聚复合物的优越性。同样重要的是，脂质涂层消除了向转染溶液中引入过量游离 PEI 链以获得更高效率的必要性，从而生成没有细胞毒性迹象的脂多聚复合物。结论 用赖氨酰化和组氨酰化胆固醇脂质对聚合复合物进行非共价修饰，可以同时提高血清条件下基因递送的效率并降低毒性，为骨髓干细胞的基因修饰带来巨大前景。
Background Effective gene transfection without serum deprivation is a prerequisite for successful stem cell-based gene therapy. Polyethylenimine (PEI) is an efficient nonviral gene vector, but its application has been hindered by serum sensitivity and severe cytotoxicity. Methods To solve this problem, a new family of lipopolyplexes was developed by coating PEI/DNA polyplexes with three serum-resistant cationic lipids, namely, lysinylated, histidylated, and arginylated cholesterol. The physical properties, transfection efficiency, cellular uptake, subcellular distribution, and cytotoxicity of the lipopolyplexes was investigated. Results The outer coat composed of lysinylated or histidylated cholesterol remarkably improved the transfection efficiency of the polyplex with a low PEI/DNA ratio of 2 in the presence of serum. The resulting lysinylated and histidylated cholesterol lipopolyplexes were even more efficient than the best performing polyplex with a high PEI/DNA ratio of 10. Results from cellular uptake and subcellular distribution studies suggest that their higher transfection efficiency may result from accelerated DNA nuclear localization. The superiority of the lipopolyplexes over the best performing polyplex was also confirmed by delivering the therapeutic gene, hVEGF165. Equally importantly, the lipid coating removed the necessity of introducing excess free PEI chains into the transfection solution for higher efficiency, generating lipopolyplexes with no signs of cytotoxicity. Conclusion Noncovalent modification of polyplexes with lysinylated and histidylated cholesterol lipids can simultaneously improve efficiency and reduce the toxicity of gene delivery under serum conditions, showing great promise for genetic modification of bone marrow stem cells.