The proper strategy to compress and protect plasmid DNA in the Pluronic L64-electropulse system for enhanced intramuscular gene delivery
The proper strategy to compress and protect plasmid DNA in the Pluronic L64-electropulse system for enhanced intramuscular gene delivery
复制标题
DOI：
10.1093/rb/rby028
复制
发表时间：
2019-10-01
影响因子：
6.7
通讯作者：
Wang, Gang
中科院分区：
文献类型：
--
作者：
He, Yutong;Liu, Yili;Wang, Gang
肌内表达功能蛋白是一种有前途的治疗策略。在此之前，我们通过结合Pluronic L 64和优化的电脉冲开发了一种肌内基因递送方法，这是迄今为止最有效的方法之一。然而，该方法中的质粒DNA（pDNA）未被压缩，使得它们在体内不稳定且效率低。我们认为，适当的压缩的pDNAs的适当的材料应促进基因表达在这个L 64-电脉冲系统。在这里，我们报告了我们发现的这样一种物质，表没食子儿茶素没食子酸酯（EGCG），一种天然化合物在绿色茶，它可以压缩和保护pDNAs和显着增加肌内基因表达的L 64-电脉冲系统。同时，我们发现聚乙烯亚胺（PEI）也可以轻微提高外源基因的表达在最佳的程序。通过分析EGCG和PEI之间的特征差异，我们得出结论，对于基于肌肉的基因递送，具有对核酸的强亲和力和/或适合于基因递送的其他特性的带负电荷的材料（如EGCG）是比阳离子材料（如PEI）更好的替代品。结果表明，材料/pDNA复合物有利于肌内基因递送/表达的关键原则是保持复合物带负电荷。这项概念验证研究显示了压缩pDNA的突破，并为开发更有效的肌内基因递送系统用于治疗应用提供了原则和策略。
Intramuscular expression of functional proteins is a promising strategy for therapeutic purposes. Previously, we developed an intramuscular gene delivery method by combining Pluronic L64 and optimized electropulse, which is among the most efficient methods to date. However, plasmid DNAs (pDNAs) in this method were not compressed, making them unstable and inefficient in vivo. We considered that a proper compression of pDNAs by an appropriate material should facilitate gene expression in this L64-electropulse system. Here, we reported our finding of such a material, Epigallocatechin gallate (EGCG), a natural compound in green teas, which could compress and protect pDNAs and significantly increase intramuscular gene expression in the L64-electropulse system. Meanwhile, we found that polyethylenimine (PEI) could also slightly improve exogenous gene expression in the optimal procedure. By analysing the characteristic differences between EGCG and PEI, we concluded that negatively charged materials with strong affinity to nucleic acids and/or other properties suitable for gene delivery, such as EGCG, are better alternatives than cationic materials (like PEI) for muscle-based gene delivery. The results revealed that a critical principle for material/pDNA complex benefitting intramuscular gene delivery/expression is to keep the complex negatively charged. This proof-of-concept study displays the breakthrough in compressing pDNAs and provides a principle and strategy to develop more efficient intramuscular gene delivery systems for therapeutic applications.