The effect of cholesterol domains on PEGylated liposomal gene delivery in vitro.
The effect of cholesterol domains on PEGylated liposomal gene delivery in vitro.
复制标题
DOI：
10.4155/tde.11.13
复制
发表时间：
2011-04
影响因子：
4.2
通讯作者：
Anchordoquy TJ
中科院分区：
文献类型：
--
作者：
Xu L;Wempe MF;Anchordoquy TJ
聚乙二醇化组分已被广泛用于减少血清中的颗粒聚集并延长基于脂质和聚合物的基因递送系统的循环寿命。然而，已知PEG化干扰细胞相互作用和细胞内运输，导致生物活性降低。在本研究中，胆固醇结构域对PEG化脂质体介导的基因递送的影响通过PEG化具有和不具有胆固醇结构域的制剂以及通过改变PEG在颗粒表面上的位置（即，在域中或从域中排除）。用PEG-胆固醇或PEG-二酰基脂质配制的脂质复合物用于转染各种细胞系，包括人和小鼠癌细胞。脂质体的细胞摄取也进行了定量，并与转染结果进行了比较。我们的发现与先前的工作一致，表明PEG化降低了转染率;然而，将PEG掺入胆固醇结构域的制剂没有表现出这种不利影响。在一些细胞系中，PEG掺入结构域实际上增加了转染率，尽管没有增强细胞摄取。这些结果表明，细胞内运输的不利改变，是聚乙二醇化的结果，可以避免通过利用传递车辆，允许PEG分区到胆固醇结构域。
PEGylated components have been widely used to reduce particle aggregation in serum and extend circulation lifetime for lipid- and polymer-based gene-delivery systems. However, PEGylation is known to interfere with cell interaction and intracellular trafficking, resulting in decreased biological activity. In the present study, the effect of cholesterol domains on PEGylated liposome-mediated gene delivery was evaluated by PEGylating formulations with and without a cholesterol domain, and also by altering the location of PEG on the particle surface (i.e., within or excluded from the domain). Lipoplexes formulated with PEG–cholesterol or PEG–diacyl lipid were used to transfect various cell lines, including human and mouse cancer cells. Cellular uptake of lipoplexes was also quantified and compared with the transfection results. Our findings are consistent with previous work demonstrating that PEGylation reduces transfection rates; however, formulations in which PEG was incorporated into the cholesterol domain did not exhibit this detrimental effect. In some cell lines, the incorporation of PEG into the domain actually increased transfection rates, despite no enhancement of cellular uptake. These results suggest that the adverse alterations in intracellular trafficking that are a consequence of PEGylation may be avoided by utilizing delivery vehicles that allow PEG to partition into a cholesterol domain.