Enhancement of airway gene transfer by DNA nanoparticles using a pH-responsive block copolymer of polyethylene glycol and poly-L-lysine.
Enhancement of airway gene transfer by DNA nanoparticles using a pH-responsive block copolymer of polyethylene glycol and poly-L-lysine.
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DOI：
10.1016/j.biomaterials.2011.11.080
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发表时间：
2012-03
期刊：
影响因子：
14
通讯作者：
Hanes, Justin
中科院分区：
文献类型：
--
作者：
Boylan, Nicholas J.;Kim, Anthony J.;Suk, Jung Soo;Adstamongkonkul, Pichet;Simons, Brian W.;Lai, Samuel K.;Cooper, Mark J.;Hanes, Justin
高度致密的DNA纳米颗粒由质粒DNA单分子与聚乙二醇和聚l -赖氨酸嵌段共聚物（PEG-CK30）致密组成，在人类基因治疗临床试验中显示出相当大的前景，但可能不太能够转染缺乏表面核蛋白的细胞。为了解决这个潜在的缺点，我们制定了ph响应DNA纳米颗粒，通过核蛋白独立途径介导基因转移。聚l -组氨酸插入聚乙二醇和聚l -赖氨酸之间，形成三嵌段共聚物体系PEG- ch12k18。聚l -组氨酸的加入将PEG-CH12K18的缓冲能力提高到与支链聚乙烯亚胺相当的水平。PEG-CH12K18将DNA压缩成棒状DNA纳米颗粒，具有与PEG-CK30 DNA纳米颗粒相似的形态和胶体稳定性。PEG-CH12K18 DNA纳米颗粒通过网格蛋白依赖的内吞机制和内溶酶体加工进入缺乏表面核蛋白的人支气管上皮细胞（BEAS-2B）。尽管通过降解内溶酶体途径运输，PEG-CH12K18 DNA纳米颗粒在体外的基因转移比PEG-CK30 DNA纳米颗粒提高了约20倍，在体内的基因转移到BALB/c小鼠的肺气道中提高了约3倍，同时保持了良好的毒性特征。这些结果代表了合理开发基于高度致密DNA纳米颗粒的高效肺基因传递平台的重要一步。
Highly compacted DNA nanoparticles, composed of single molecules of plasmid DNA compacted with block copolymers of polyethylene glycol and poly-L-lysine (PEG-CK30), have shown considerable promise in human gene therapy clinical trials in the nares, but may be less capable of transfecting cells that lack surface nucleolin. To address this potential shortcoming, we formulated pH-responsive DNA nanoparticles that mediate gene transfer via a nucleolin-independent pathway. Poly-L-histidine was inserted between PEG and poly-L-lysine to form a triblock copolymer system, PEG-CH12K18. Inclusion of poly-L-histidine increased the buffering capacity of PEG-CH12K18 to levels comparable with branched polyethyleneimine. PEG-CH12K18 compacted DNA into rod-shaped DNA nanoparticles with similar morphology and colloidal stability as PEG-CK30 DNA nanoparticles. PEG-CH12K18 DNA nanoparticles entered human bronchial epithelial cells (BEAS-2B) that lack surface nucleolin by a clathrin-dependent endocytic mechanism followed by endo-lysosomal processing. Despite trafficking through the degradative endo-lysosomal pathway, PEG-CH12K18 DNA nanoparticles improved the in vitro gene transfer by ~ 20-fold over PEG-CK30 DNA nanoparticles, and in vivo gene transfer to lung airways in BALB/c mice by ~ 3-fold, while maintaining a favorable toxicity profile. These results represent an important step toward the rational development of an efficient gene delivery platform for the lungs based on highly compacted DNA nanoparticles.
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影响因子：
4.8
作者：
Liu, G;Li, DS;Cooper, MJ
通讯作者：
Cooper, MJ
影响因子：
5.1
作者：
Fasbender, A;Zabner, J;Welsh, MJ
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46.9
作者：
Hyde, Stephen C.;Pringle, Ian A.;Gill, Deborah R.
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Gill, Deborah R.
影响因子：
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作者：
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Hyde, SC
影响因子：
4.2
作者：
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通讯作者：
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