Transfection study using multicellular tumor spheroids for screening non-viral polymeric gene vectors with low cytotoxicity and high transfection efficiencies
Transfection study using multicellular tumor spheroids for screening non-viral polymeric gene vectors with low cytotoxicity and high transfection efficiencies
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DOI：
10.1016/j.jconrel.2007.05.012
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发表时间：
2007-08-16
影响因子：
10.8
通讯作者：
Kataoka, Kazurlori
中科院分区：
文献类型：
--
作者：
Han, Muri;Bae, Younsoo;Kataoka, Kazurlori
由质粒DNA和聚阳离子组成的复合物作为基因转移载体受到了广泛关注。为了有效的基因治疗，开发了在侧链中具有不同多胺结构的聚阳离子以确保其对内体逃逸的缓冲能力，并且开发了其PEG化嵌段共聚物以增加其稳定性和生物相容性。利用人肝癌HuH-7细胞的三维多细胞肿瘤球体，阐明了聚阳离子的化学结构及其PEG化对转染和细胞毒性的影响。多细胞肿瘤球体（MCTS）模型揭示了复合物胶束转染的各种特征，这些特征在传统的单层培养中难以观察到，在三维微环境下的细胞毒性和转染基因表达的时间依赖性行为。通过使用该系统，来自在侧链中具有N-（2-氨乙基）-2-氨乙基的聚（乙二醇）-b-聚（N-取代的天冬酰胺）共聚物（PEG-b-P[Asp（DET）]聚复合物胶束）的聚复合物胶束被证明实现高转染效率以及低细胞毒性，这两者对于成功的体内基因递送是关键性质。C）2007 Elsevier B. V.保留所有权利。
Polyplexes consisting of plasmid DNA and polycations have received much attention as promising vectors for gene transfer. For effective gene therapy, polycations with different polyamine structures in the side chain were developed to ensure their buffering capacity for endosomal escape, and their PEGylated block copolymers were developed to increase their stability and biocompatibility. The effects of the chemical structures of polycations and their PEGylation on transfection and cytotoxicity were elucidated by use of a three-dimensional multicellular tumor spheroid of human hepatoma HuH-7 cells. Various features of transfection with polyplex micelles, which have been hard to observe in conventional monolayer cultures, were revealed by the multicellular tumor spheroid (MCTS) model in terms of cytotoxicity and time-dependent behaviors of transfected gene expression under three-dimensional microenvironments. By using this system, the polyplex micelle from poly(ethylene glycol)-b-poly(N-substituted asparagine) copolymers having the N-(2-aminoethyl)-2-aminoethyl group in the side chain (PEG-b-P[Asp(DET)] polyplex micelle) was proved to achieve high transfection efficiencies as well as low cytotoxicity, both of which are critical properties for successful in vivo gene delivery. C) 2007 Elsevier B.V. All rights reserved.