Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer vaccines: Programmed endosomal escape and dissociation
Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer vaccines: Programmed endosomal escape and dissociation
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DOI：
10.1016/j.jconrel.2010.01.012
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发表时间：
2010-05-10
影响因子：
10.8
通讯作者：
Harashima, Hideyoshi
中科院分区：
文献类型：
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作者：
Akita, Hidetaka;Kogure, Kentaro;Harashima, Hideyoshi
我们之前开发了八精氨酸（R8）修饰的脂质被膜型纳米颗粒用于siRNA递送（R8-MEND）。在此，我们报告了它们的离体siRNA递送至原代小鼠骨髓来源的树突状细胞（BMDC）以用作癌症疫苗的潜在用途。siRNA的细胞内运输的定量成像分析显示，解离过程以及内体逃逸的速率限制了通过水合方法（R8-MENDhydo）制备的原型R8-MEND的siRNA效率。成功的内体逃逸是通过使用pH依赖性融合肽（GALA）修饰的脂质混合物，优化内体融合。此外，用于制备纳米颗粒的改进方案，混合5iRNA/STR-R8复合物和小单层囊泡（R8/GALA-MENDSUV），导致更均匀、更小的粒度，并导致更有效的细胞内解离。细胞因子信号转导抑制因子1（SOCS1）是BMDCs中免疫应答的负反馈调节因子，其基因敲低导致STAT1磷酸化增强和促炎细胞因子的产生。此外，SOCS1沉默的BMDC在抑制肿瘤生长方面更有效。总的来说，这些结果表明，负载在R8/GALA-MENDSUV中的siRNA有效地抑制内源性基因表达，从而增强基于树突状细胞的疫苗在体内的效力。(C)2010 Elsevier B.V.保留所有权利。
We previously developed octaarginine (R8)-modified lipid envelope-type nanoparticles for siRNA delivery (R8-MEND). Herein, we report on their ex vivo siRNA delivery to primary mouse bone marrow-derived dendritic cells (BMDCs) for potential use as a cancer vaccine. Quantitative imaging analysis of the intracellular trafficking of siRNA revealed that the dissociation process, as well as the rate of endosomal escape limits the siRNA efficiency of the prototype R8-MEND, prepared by the hydration method (R8-MENDhydo). Successful endosomal escape was achieved by using a pH-dependent fusogenic peptide (GALA) modified on a lipid mixture that was optimized for endosomal fusion. Furthermore, a modified protocol for the preparation of nanoparticles, mixing the 5iRNA/STR-R8 complex and small unilamellar vesicles (R8/GALA-MENDSUV), results in a more homogenous, smaller particle size, and results in a more efficient intracellular dissociation. Gene knockdown of the suppressor of cytokine signaling 1 (SOCS1), a negative-feedback regulator of the immune response in BMDCs resulted in an enhanced phosphorylation of STAT1, and the production of proinflammatory cytokines. Moreover, SOCS1-silenced BMDCs were more potent in suppressing tumor growth. Collectively, these results show that siRNA loaded in R8/GALA-MENDSUV efficiently suppresses endogenous gene expression and consequently enhances dendritic cell-based vaccine potency in vivo. (C) 2010 Elsevier B.V. All rights reserved.