Extracellular vesicle (ECV)-modified polyethylenimine (PEI) complexes for enhanced siRNA delivery in vitro and in vivo.
Extracellular vesicle (ECV)-modified polyethylenimine (PEI) complexes for enhanced siRNA delivery in vitro and in vivo.
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DOI：
10.1016/j.jconrel.2019.12.032
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发表时间：
2019-12
期刊：
影响因子：
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通讯作者：
Petro Zhupanyn;A. Ewe;Thomas Büch;A. Malek;Phil Rademacher;C. Müller;Anja Reinert;Y. Jaimes;A. Aigner
中科院分区：
文献类型：
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作者：
Petro Zhupanyn;A. Ewe;Thomas Büch;A. Malek;Phil Rademacher;C. Müller;Anja Reinert;Y. Jaimes;A. Aigner
细胞外囊泡（ECV）是细胞分泌的膜颗粒，参与细胞间信号传导和细胞间通讯。通过运输各种生物大分子，ECV和特别是外来体在各种（病理）生理过程中是相关的。ECV也由癌细胞释放，并且可以赋予促肿瘤发生作用。它们的靶细胞向性、对增殖速率的影响、在血液中的天然稳定性和免疫耐受性使得ECV作为递送载体特别令人感兴趣。聚乙烯亚胺（PEI）是能够与小RNA分子（包括siRNA或antimiR）形成非共价复合物的线性或支化聚合物，本研究首次探索了基于PEI的纳米颗粒与来自不同细胞系的天然存在的ECV的组合，用于递送小RNA。通过电子显微镜对比单独的ECV或复合物分析ECV修饰的PEI/siRNA复合物。在功能方面，我们证明了PEI/siRNA复合物在用ECV修饰后的敲低功效和储存稳定性增加。这通过靶向Survivin的ECV修饰的PEI/siRNA复合物增强的肿瘤细胞抑制来证实。用细胞内化的各种抑制剂预处理揭示了PEI/siRNA复合物在其ECV修饰后的细胞摄取机制和生物活性的改变。将我们的研究扩展到针对miR-155或miR-1246的PEI复合的antimiR，基于直接miRNA靶基因的去抑制，ECV修饰的复合物中的剂量依赖性细胞和分子效应增强。观察到来自不同细胞系的ECV之间关于其增强PEI/siRNA功效的能力的差异，与用于转染的靶细胞系无关。最后，在携带s.c. PC 3前列腺癌异种移植物显示，基于深刻的靶基因敲低，在用ECVPC 3修饰的PEI/siSurvivin复合物处理后，肿瘤生长受到显著抑制。我们得出结论，ECV修饰增强了PEI为基础的复合物的活性，通过改变关键的物理化学和生物纳米粒子的性能。
Extracellular vesicles (ECVs) are secreted cell-derived membrane particles involved in intercellular signaling and cell-cell communication. By transporting various bio-macromolecules, ECVs and in particular exosomes are relevant in various (patho-) physiological processes. ECVs are also released by cancer cells and can confer pro-tumorigenic effects. Their target cell tropism, effects on proliferation rates, natural stability in blood and immunotolerance makes ECVs particularly interesting as delivery vehicles.Polyethylenimines (PEIs) are linear or branched polymers which are capable of forming non-covalent complexes with small RNA molecules including siRNAs or antimiRs, for their delivery in vitro and in vivo.This study explores for the first time the combination of PEI-based nanoparticles with naturally occurring ECVs from different cell lines, for the delivery of small RNAs. ECV-modified PEI/siRNA complexes are analyzed by electron microscopy vs. ECV or complex alone. On the functional side, we demonstrate increased knockdown efficacy and storage stability of PEI/siRNA complexes upon their modification with ECVs. This is paralleled by enhanced tumor cell-inhibition by ECV-modified PEI/siRNA complexes targeting Survivin. Pre-treatment with various inhibitors of cellular internalization reveals alterations in cellular uptake mechanisms and biological activities of PEI/siRNA complexes upon their ECV modification. Extending our studies towards PEI-complexed antimiRs against miR-155 or miR-1246, dose-dependent cellular and molecular effects are enhanced in ECV-modified complexes, based on the de-repression of direct miRNA target genes. Differences between ECVs from different cell lines are observed regarding their capacity of enhancing PEI/siRNA efficacies, independent of the target cell line for transfection. Finally, an in vivo therapy study in mice bearing s.c. PC3 prostate carcinoma xenografts reveals marked inhibition of tumor growth upon treatment with ECVPC3-modified PEI/siSurvivin complexes, based on profound target gene knockdown. We conclude that ECV-modification enhances the activity of PEI-based complexes, by altering pivotal physicochemical and biological nanoparticle properties.