深入了解脂质纳米颗粒的细胞摄取机制和小RNA的细胞内释放。
Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs.
作者信息
Yu Bo, Wang Xinmei, Zhou Chenguang, Teng Lesheng, Ren Wei, Yang Zhaogang, Shih Chih-Hsin, Wang Tianyou, Lee Robert J, Tang Suoqin, Lee L James
机构信息
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA.
出版信息
Pharm Res. 2014 Oct;31(10):2685-95. doi: 10.1007/s11095-014-1366-7. Epub 2014 Apr 17.
PURPOSE
Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and microRNA (miRNA).
METHOD
In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.
RESULTS
We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.
CONCLUSIONS
Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.
目的
了解细胞摄取和细胞内释放机制将有助于更好地设计用于递送诸如小干扰RNA（siRNA）和微小RNA（miRNA）等核酸的纳米载体。
方法
在本研究中，我们通过将荧光标记的siRNA和分子信标（MB）共包封在四种不同配方的阳离子脂质纳米颗粒（LNP）中，研究了siRNA的细胞药代动力学。一种miRNA模拟物也被用作研究细胞药代动力学的探针，其与RNA干扰活性具有良好的相关性。
结果
我们试图基于二油酰基三甲基氯化铵（DOTMA）和二油酰基二甲基氯化铵（DODMA）的组合找到最佳的LNP配方。当DOTMA/DODMA比例为5/40时，含有荧光素酶siRNA的LNP产生了最高的基因沉默活性。DOTMA/DODMA的卓越效力可归因于更高的摄取率以及摄取后促进siRNA从内体释放的能力提高。
结论
我们的研究结果可能为RNA干扰转染途径提供新的见解，并对阳离子LNP设计具有启示意义。
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