Effective Gene Delivery Using Stimulus-Responsive Catiomer Designed with Redox-Sensitive Disulfide and Acid-Labile Imine Linkers
Effective Gene Delivery Using Stimulus-Responsive Catiomer Designed with Redox-Sensitive Disulfide and Acid-Labile Imine Linkers
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使用由氧化还原敏感的二硫化物和酸不稳定的亚胺接头设计的刺激响应型阳离子异构体进行有效的基因递送。
DOI：
10.1021/bm2017355
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发表时间：
2012-04-01
影响因子：
6.2
通讯作者：
Shi, Donglu
中科院分区：
文献类型：
--
作者：
Cai, Xiaojun;Dong, Chunyan;Shi, Donglu
研制了一种双刺激响应型mpeg-SS-PLL(15)-戊二醛STAR(mpeg-SS-PLL(15)-STAR)阳离子聚合物，并对其进行了生物学评价。阳离子聚合物系统结合了氧化还原敏感的外部聚乙二醇壳的去除和酸诱导的内体间隙的逃逸。聚乙二醇壳去除的设计原理是在肿瘤相关谷胱甘肽(GSH)浓度存在的情况下增加细胞内对mpeg-SS-PLL(15)-star/DNA复合体的摄取，而酸诱导的解离是为了在成功内化到靶细胞后加速基因有效载荷的释放。在10 mM GSH存在下，复合体的大小变化表明，在细胞内存在于肿瘤微环境中的氧化还原条件下，刺激诱导的外部聚乙二醇层脱落。动态激光光散射实验表明，在内体pH条件下，mpeg-SS-PLL(15)-STAR/DNA复合体快速失稳，随后通过琼脂糖凝胶电泳法促进包裹的DNA的有效释放。以编码绿色荧光蛋白的pEGFP-C1质粒DNA和编码荧光素酶的PGL-3质粒DNA为报告基因进行的生物学效果评价表明，该阳离子聚合物对293T细胞的转染率与传统的基于聚乙烯亚胺(bPEI-25k)的基因递送系统相当。这些实验结果表明，mpeg-SS-PLL(15)-STAR具有高DNA结合能力、低细胞毒性和高转染率的特点，是未来非病毒基因传递应用的一种有前途的设计。
A dual stimulus-responsive mPEG-SS-PLL(15)-glutaraldehyde star (mPEG-SS-PLL(15)-star) catiomer is developed and biologically evaluated. The catiomer system combines redox-sensitive removal of an external PEG shell with acid-induced escape from the endosomal compartment. The design rationale for PEG shell removal is to augment intracellular uptake of mPEG-SS-PLL(15)-star/DNA complexes in the presence of tumor-relevant glutathione (GSH) concentration, while the acid-induced dissociation is to accelerate the release of genetic payload following successful internalization into targeted cells. Size alterations of complexes in the presence of 10 mM GSH suggest stimulus-induced shedding of external PEG layers under redox conditions that intracellularly present in the tumor microenvironment. Dynamic laser light scattering experiments under endosomal pH conditions show rapid destabilization of mPEG-SS-PLL(15)-star/DNA complexes that is followed by facilitating efficient release of encapsulated DNA, as demonstrated by agarose gel electrophoresis. Biological efficacy assessment using pEGFP-C1 plasmid DNA encoding green fluorescence protein and pGL-3 plasmid DNA encoding luciferase as reporter genes indicate comparable transfection efficiency of 293T cells of the catiomer with a conventional polyethyleneimine (bPEI-25k)-based gene delivery system. These experimental results show that mPEG-SS-PLL(15)-star represents a promising design for future nonviral gene delivery applications with high DNA binding ability, low cytotoxicity, and high transfection efficiency.