Supramolecular control of polyplex dissociation and cell transfection: Efficacy of amino groups and threading cyclodextrins in biocleavable polyrotaxanes
Supramolecular control of polyplex dissociation and cell transfection: Efficacy of amino groups and threading cyclodextrins in biocleavable polyrotaxanes
复制标题
DOI：
10.1016/j.jconrel.2008.07.011
复制
发表时间：
2008-10-21
影响因子：
10.8
通讯作者：
Harashima, Hideyoshi
中科院分区：
文献类型：
--
作者：
Yamashita, Atsushi;Kanda, Daizo;Harashima, Hideyoshi
利用生物可裂解聚轮烷进行基因传递的一种新策略是将二甲氨基乙基修饰的α-环糊精(DMAE-α-CDs)通过二硫键连接到以苄氧羰基-L酪氨酸(DMAE-SS-Prx)为端基的聚乙二醇链(DMAE-SS-Prx)上，该策略涉及到与反多阴离子形成稳定的多离子络合物(Polyplex)，并伴随着DMAE-SS-Prx的超分子解离。在本研究中，我们制备了具有不同数量的线性化α-CD和氨基(DMAE)的生物可降解聚轮烷，以提高DMAE-SS-PRXs的转染活性。29DMAE-α18-SS-PRX的α-CD分子数为18个，氨基数目为29个，与其他PRX相比，具有较高的转染率。DMAE-SS-PRXs的转染活性似乎与PDNA从这些复合体中释放的效果有关，这是由多轮烷载体中α-CD和/或氨基的数量控制的。正如预期的那样，大多数DMAE-SS-PRX聚合物只在10 mM DTT和反多聚阴离子存在的情况下释放PDNA，除了14DMAE-α18-SS-PRX，一旦DTT加入到聚合物溶液中，14DMAE-α18-SS-PRX在没有葡聚糖硫酸盐的情况下释放PDNA。与29DMAE-α18-SS-PRX相比，14DMAE-α18-SS-PRX的转染率明显降低。激光共聚焦扫描显微镜(CLSM)观察表明，14DMAE-α18-SS-PRX的特异性结果可能是由于胞浆中最解离的14DMAE-α18-SS-PRX多聚体过早释放了PDNA。因此，转染活性似乎与适当的PDNA释放时间有关。(C)2008爱思唯尔B.V.保留所有权利。
A novel strategy for gene delivery using biocleavable polyrotaxanes, in which dimethylaminoethyl-modified alpha-cyclodextrins (DMAE-alpha-CDs) are threaded onto a poly(ethylene glycol) (PEG) chain capped with benzyloxycarbonyl-L-tyrosine via disulfide linkages (DMAE-SS-PRX), involves the formation of a stable polyion complex (polyplex) against a counter polyanion and the intracellular plasmid DNA (pDNA) release from the polyplex accompanied by the supramolecular dissociation of DMAE-SS-PRXs. In this study, we prepared biocleavable polyrotaxanes with different numbers of threading alpha-CD and amino (DMAE) groups to enhance the transfection activity of DMAE-SS-PRXs. 29DMAE-alpha 18-SS-PRX, in which the numbers of alpha-CD molecules and amino groups were 18 and 29 respectively, exhibited a high transfection activity compared with other PRXs. The transfection activity of DMAE-SS-PRXs seems to be related to the efficacy of pDNA release from those polyplexes, which was controlled by the number of alpha-CD and/or amino groups in the polyrotaxane carrier. Most of the DMAE-SS-PRX polyplexes released the pDNA only in the presence of both 10 mM DTT and of the counter-polyanion, as expected, except for 14DMAE-alpha 18-SS-PRX, which released pDNA in the absence of dextran sulfate once the DTT had been added to the polyplex solution. The transfection activity of 14DMAE-alpha 18-SS-PRX was significantly lower than that of 29DMAE-alpha 18-SS-PRX regardless of the above features. Confocal laser scanning microscopic (CLSM) observation suggested that the specific result for 14DMAE-alpha 18-SS-PRX might be due to a premature release of pDNA from the most dissociative 14DMAE-alpha 18-SS-PRX polyplex in the cytosol. Therefore, transfection activity seems to be related to an appropriate timing of pDNA release. (C) 2008 Elsevier B.V. All rights reserved.