Effect of inserted spacer in hepatic cell-penetrating multifunctional peptide component on the DNA intracellular delivery of quaternary complexes based on modular design.
Effect of inserted spacer in hepatic cell-penetrating multifunctional peptide component on the DNA intracellular delivery of quaternary complexes based on modular design.
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基于模块化设计的肝细胞穿透多功能肽成分中插入间隔基对四元复合物DNA细胞内递送的影响
DOI：
10.2147/ijn.s115381
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发表时间：
2016
影响因子：
8
通讯作者：
Du Z
中科院分区：
文献类型：
--
作者：
Zhang L;Li Z;Sun F;Xu Y;Du Z
基于分子间的非共价键自组装，构建了一种安全、高效的四元基因传递系统（简称Q复合物）。该系统通过阳离子脂质体、多功能肽和DNA的合作和竞争相互作用来配制，然后在三元复合物的表面上涂覆透明质酸。多功能肽由两个功能结构域组成：穿透肝肿瘤靶向细胞部分（KRPTMRFRYTWNPMK）和包装基因序列（聚精氨酸16）。进一步研究了在多功能肽的两个结构域之间插入间隔子对Q-复合物细胞内转染的影响。实验结果表明，在间隔元件中组装不同肽的制剂具有不同的细胞间途径和转染效率。在不存在间隔元件（Pa）的情况下含有肽的Q-复合物在所有样品中显示出最高的基因表达。含有具有不可切割间隔区GA（Pc）的肽的Q-复合物没有细胞内核酸递送能力，而具有可切割间隔区RVRR（Pd）的那些显示出中等的转染活性。这些结果表明，插入多功能肽中的不同间隔区在体外DNA转染效率中起重要作用。原子力显微镜图像显示，三元配合物（LPcD）和Q-配合物（HLcPD）的形貌为层状晶体，而其他纳米配合物的形貌为球形。圆二色性分析表明，与游离态相比，含间隔基GA的纳米复合物中肽的构型发生了变化，而不含间隔基的纳米复合物中的Pa构型与其游离态一致。本研究有助于对Q-配合物结构的理解，进一步有效的修饰正在进行中。
A safe and efficient quaternary gene delivery system (named Q-complexes) was constructed based on self-assembly of molecules through noncovalent bonds. This system was formulated through the cooperation and competing interactions of cationic liposomes, multifunctional peptides, and DNA, followed by coating hyaluronic acid on the surface of the ternary complexes. The multifunctional peptide was composed of two functional domains: penetrating hepatic tumor-targeted cell moiety (KRPTMRFRYTWNPMK) and a wrapping gene sequence (polyarginine 16). The effect of spacer insertion between the two domains of multifunctional peptide on the intracellular transfection of Q-complexes was further studied. Experimental results showed that the formulations assembled with various peptides in the spacer elements possessed different intercellular pathways and transfection efficiencies. The Q-complexes containing peptide in the absence of spacer element (Pa) showed the highest gene expression among all samples. The Q-complexes containing peptides with a noncleavable spacer GA (Pc) had no ability of intracellular nucleic acid delivery, whereas those with a cleavable spacer RVRR (Pd) showed moderate transfection activity. These results demonstrated that the different spacers inserted in the multifunctional peptide played an important role in in vitro DNA transfection efficiency. Atomic force microscopy images showed that the morphologies of ternary complexes (LPcD) and Q-complexes (HLcPD) were crystal lamellas, whereas those of other nanocomplexes were spheres. Circular dichroism showed the changed configuration of peptide with spacer GA in nanocomplexes compared with that of its free state, whereas the Pa configuration without spacer in nanocomplexes was consistent with that of its free state. The present study contributed to the structural understanding of Q-complexes, and further effective modification is in progress.