Role of polymeric endosomolytic agents in gene transfection: a comparative study of poly(L-lysine) grafted with monomeric L-histidine analogue and poly(L-histidine).
Role of polymeric endosomolytic agents in gene transfection: a comparative study of poly(L-lysine) grafted with monomeric L-histidine analogue and poly(L-histidine).
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DOI：
10.1021/bm500843r
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发表时间：
2014-10-13
影响因子：
6.2
通讯作者：
Bae, You Han
中科院分区：
文献类型：
--
作者：
Hwang, Hee Sook;Hu, Jun;Na, Kun;Bae, You Han
内体包裹是有效基因表达以及细胞内化、DNA释放和核输入必须克服的主要障碍之一。在聚阳离子聚合物中引入ph敏感的可电离基团以提高基因转移效率已被证明是一种有效的方法；然而，在聚合物和单体形式中引入相同数量的可电离基团的比较研究尚未报道。在这项研究中，我们制备了两种类型的组氨酸接枝聚l-赖氨酸（PLL），一种是聚l-组氨酸（PLL-g- phis）的堆叠形式，另一种是具有相同数量咪唑基团的单l-组氨酸（PLL-g- mhis）。这两种类型的组氨酸移植PLL， PLL-g- phis和PLL-g- mhis，在溶血活性、细胞摄取、内化和转染效率方面表现出深刻的差异。cy3标记的PLL-g-PHis内化后在细胞核中显示出强烈的荧光，并且在pH变化时也观察到PLL-g-PHis具有较高的溶血活性。来自PHis的咪唑基团的排列也提供了比mHis更高的基因表达，因为它能够逃离核内体。mHis或PHis接枝降低了PLL的细胞毒性，并通过改变可用于基因凝聚的游离ε-胺的数量改变了PLL的细胞摄取速率。YOYO1-pDNA强度测定的PLL-g-PHis/pDNA亚细胞定位在细胞核内最高，而染色酸性区室的溶酶追踪器亚细胞定位在这些聚合物中最低。因此，聚合组氨酸的排列显示出能够逃离核内体并触发多聚体快速释放到细胞质中，从而导致更多的pDNA可用于易位到细胞核并增强基因表达。
Endosomal entrapment is one of the main barriers that must be overcome for efficient gene expression along with cell internalization, DNA release, and nuclear import. Introducing pH-sensitive ionizable groups into the polycationic polymers to increase gene transfer efficiency has proven to be a useful method; however, a comparative study of introducing equal numbers of ionizable groups in both polymer and monomer forms, has not been reported. In this study, we prepared two types of histidine-grafted poly(l-lysine) (PLL), a stacking form of poly(l-histidine) (PLL-g-PHis) and a mono- l-histidine (PLL-g-mHis) with the same number of imidazole groups. These two types of histidine-grafted PLL, PLL-g-PHis and PLL-g-mHis, showed profound differences in hemolytic activity, cellular uptake, internalization, and transfection efficiency. Cy3-labeled PLL-g-PHis showed strong fluorescence in the nucleus after internalization, and high hemolytic activity upon pH changes was also observed from PLL-g-PHis. The arrangement of imidazole groups from PHis also provided higher gene expression than mHis due to its ability to escape the endosome. mHis or PHis grafting reduced the cytotoxicity of PLL and changed the rate of cellular uptake by changing the quantity of free ε-amines available for gene condensation. The subcellular localization of PLL-g-PHis/pDNA measured by YOYO1-pDNA intensity was highest inside the nucleus, while the lysotracker, which stains the acidic compartments was lowest among these polymers. Thus, the polymeric histidine arrangement demonstrate the ability to escape the endosome and trigger rapid release of polyplexes into the cytosol, resulting in a greater amount of pDNA available for translocation to the nucleus and enhanced gene expression.
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影响因子：
14
作者：
Boylan, Nicholas J.;Kim, Anthony J.;Suk, Jung Soo;Adstamongkonkul, Pichet;Simons, Brian W.;Lai, Samuel K.;Cooper, Mark J.;Hanes, Justin
通讯作者：
Hanes, Justin
DOI：
10.1016/j.jconrel.2012.06.036
发表时间：
2012-12-28
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Kang HC;Huh KM;Bae YH
通讯作者：
Bae YH
影响因子：
14
作者：
Kang, Han Chang;Kang, Ho-Jung;Bae, You Han
通讯作者：
Bae, You Han
影响因子：
4.7
作者：
Chang, Kai-Ling;Higuchi, Yuriko;Hashida, Mitsuru
通讯作者：
Hashida, Mitsuru
影响因子：
6.2
作者：
Griffiths, PC;Paul, A;Duncan, R
通讯作者：
Duncan, R