Pyridylthiourea-grafted polyethylenimine offers an effective assistance to siRNA-mediated gene silencing in vitro and in vivo
Pyridylthiourea-grafted polyethylenimine offers an effective assistance to siRNA-mediated gene silencing in vitro and in vivo
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DOI：
10.1016/j.jconrel.2011.10.007
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发表时间：
2012-02-10
影响因子：
10.8
通讯作者：
Zuber, Guy
中科院分区：
文献类型：
--
作者：
Creusat, Gaelle;Thomann, Jean-Sebastien;Zuber, Guy
基于合成干扰核酸(SiRNAs)的治疗的成功几乎完全依赖于有效、安全、最好是纳米级的递送系统，这些系统即使在高浓度下也可以很容易地制备。我们通过化学合成的方法制备了各种自组装聚合物，将siRNA包裹到细胞外稳定的多聚体中，但在感受到内体酸性时具有分解潜力。结果表明，吡啶硫脲接枝聚乙烯亚胺(Pi PEI)遵循上述原则。在体外，10 nM的siRNA浓度可使U87细胞90%以上的基因沉默，且不具有溶血活性。SiRNA/pi PEI的组装。然后研究了在高浓度下，4.5%葡萄糖溶液，pH 6.0，几个小时后得到尺寸略小于100 nm的稳定胶体溶液。将这些浓缩的siRNA多聚体注射到表达荧光素酶的人胶质母细胞瘤中，移植到裸鼠皮下，注射后4天，30%的siRNA介导的荧光素酶基因显著沉默。我们的结果在体外和体内实验中都证实了带有pH敏感拆解开关的阳离子聚合物自组装用于siRNA传递的潜力。(C)2011爱思唯尔B.V.保留所有权利。
Success of synthetic interfering nucleic acids (siRNAs)-based therapy relies almost exclusively on effective, safe and preferably nanometric delivery systems which can be easily prepared, even at high concentrations. We prepared by chemical synthesis various self-assembling polymers to entrap siRNAs into stable polyplexes outside cells but with a disassembly potential upon sensing endosomal acidity. Our results revealed that pyridylthiourea-grafted polyethylenimine (pi PEI) followed the above-mentioned principles. It led to above 90% siRNA-mediated gene silencing in vitro on U87 cells at 10 nM siRNA concentration and did not have a hemolytic activity. Assembly of siRNA/pi PEI. at high concentration was then studied and 4.5% glucose solution, pH 6.0, yielded stable colloidal solutions with sizes slightly below 100 nm for several hours. A single injection of these concentrated siRNA polyplexes into luciferase-expressing human glioblastoma tumors, which were subcutaneously xenografted into nude mice, led to a significant 30% siRNA-mediated luciferase gene silencing 4 days post-injection. Our results altogether substantiate the potential of self-assembling cationic polymers with a pH-sensitive disassembly switch for siRNA delivery in vitro and also in vivo experiments. (C) 2011 Elsevier B. V. All rights reserved.