Tailor-made ternary nanopolyplexes of thiolated trimethylated chitosan with pDNA and folate conjugated cis-aconitic amide-polyethylenimine for efficient gene delivery.
Tailor-made ternary nanopolyplexes of thiolated trimethylated chitosan with pDNA and folate conjugated cis-aconitic amide-polyethylenimine for efficient gene delivery.
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DOI：
10.1016/j.ijbiomac.2019.10.212
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发表时间：
2019-11
影响因子：
8.2
通讯作者：
Qing Liu;Zhu Jin;Wei Huang;Y. Sheng;Zhongmin Wang;Shengrong Guo
中科院分区：
文献类型：
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作者：
Qing Liu;Zhu Jin;Wei Huang;Y. Sheng;Zhongmin Wang;Shengrong Guo
为了克服细胞外/细胞内不同的基因传递障碍，通过模块化的自组装策略制备了具有电荷转化特性的肿瘤靶向和pH/氧化还原响应型三元复合体。首先，通过静电作用和二硫键的形成，合成了硫代化三甲基壳聚糖(TMC-SH)，使其与PDNA发生交联缩合，得到了氧化还原响应型基因释放的TMC-SS/PDNA二元复合体。为了进一步赋予二元聚合物肿瘤靶向性和内切/溶酶体pH触发的电荷反转特性，合成了叶酸偶联顺式-乌头酰胺-聚乙烯亚胺(FA-PEI-ACO)来屏蔽正向TMC-SS/PDNA，生成了尺寸约190 nm、表面带负电荷的FA-PEI-ACO/TMC-SS/PDNA三元聚合物。在生理pH条件下，复合体的ζ-电位稳定，在pH为5.5(内切/溶酶体pH)时，膜电位从-14 mV迅速增加到+20 mV，这是由于酸敏感的酰胺键断裂和FA-PEI-ACO层的去屏蔽所致，这可能有利于复合体的内切/溶酶体逃逸。然后，复合体可以在细胞内较高浓度的谷胱甘肽的情况下氧化还原反应释放基因。通过利用这种多重反应，在体外培养的Hela细胞中实现了对三元多聚体的显著提高的转染率。这些结果表明，新开发的复合体在基因传递方面具有潜在的应用前景。
To overcome the different extra-/intracellular barriers in gene delivery, tumor-targeted and pH/redox-responsive ternary polyplexes with charge-conversional properties were prepared through a modular self-assembly strategy. Firstly, the thiolated trimethylated chitosan (TMC-SH) was synthesized to crosslink and condense pDNA through electrostatic interaction and disulfide formation, which obtained the TMC-SS/pDNA binary polyplexes with redox-responsive gene release. To further endow the binary polyplexes with tumor targeting and endo/lysosomal pH-triggered charge-reversal properties, a folate conjugatedcis-aconitic amide-polyethylenimine (FA-PEI-AcO) was synthesized to shield the positive TMC-SS/pDNA, generating the FA-PEI-AcO/TMC-SS/pDNA ternary polyplexes with a size of ~190 nm and negative surface-charges. The ζ-potential of the polyplexes was stable at physiological pH and increased rapidly from -14 mV to + 20 mV at pH 5.5 (endo/lysosomal pH) due to the breakages of acid-liable amide bonds and the subsequent de-shielding of FA-PEI-AcO layers, which might benefit the endo/lysosomal escape of the polyplexes. Afterward, the polyplexes could redox-responsively release gene at higher intracellular concentrations of glutathione. By taking advantage of such multi-responses, significantly enhanced transfection efficiency was achievedin vitroin Hela cells for the ternary polyplexes. These results suggested that the newly developed polyplexes had potential application for gene delivery.