Milk exosomes - Natural nanoparticles for siRNA delivery
Milk exosomes - Natural nanoparticles for siRNA delivery
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DOI：
10.1016/j.canlet.2019.02.011
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发表时间：
2019-01-01
期刊：
影响因子：
9.7
通讯作者：
Gupta, Ramesh C.
中科院分区：
文献类型：
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作者：
Aqil, Farrukh;Munagala, Radha;Gupta, Ramesh C.
靶向siRNAs的基因沉默作为一种治疗包括癌症在内的各种疾病的方法具有巨大的潜力。然而，siRNA在细胞内的传递是具有挑战性的。我们使用牛乳外切体作为一种新的siRNA传递系统。首先，我们证明了外切体可以将内源性RNA有效载荷运送到受体细胞中。接下来，我们加载针对特定基因的siRNA，包括血管内皮生长因子、EGFR、AKT、MAPK和KRAS。我们利用5‘(-32)P标记的siKRAS作为示踪剂，发现外体对siRNA的负载量是可变的。我们证明了负载外切体的siRNA是稳定的，并能抵抗降解。我们的结果表明，针对靶基因的siRNAs在不同癌症中的表达水平从2倍到10倍不等。由于突变的KRAS与包括肺癌在内的各种癌症的发生有关，我们在A549细胞中测试了针对KRAS(G12S)的突变等位基因特异性siRNA。我们观察到携带siKRAS(G12S)的外切体对A549细胞具有剂量依赖的抗增殖活性。我们观察到，携带siKRAS的叶酸功能化外切体对A549肿瘤移植瘤有显著的抑制作用。综上所述，牛奶来源的外切体是一种可行的天然纳米载体，可用于运送siRNA用于抗癌治疗。
Gene-silencing with targeted siRNAs has great potential as a therapeutic approach for various diseases including cancer. However, intracellular delivery of siRNA is challenging. We used bovine milk exosomes as a novel system for siRNA delivery. First, we demonstrated that exosomes can deliver endogenous RNA payloads into recipient cells. Next, we loaded siRNA against specific genes including VEGF, EGFR, AKT, MAPK, and KRAS. We utilized 5'(-32) P-labeled siKRAS as a tracer and found exosome loading with siRNA could be variable. We demonstrated that the siRNA of loaded exosomes is stable and resist degradation. Our results indicated that siRNAs against target genes ranged from 2 to 10-fold knockdown in expression levels in various cancers. Since mutated KRAS has been implicated in the development of various cancers including lung cancer, we tested a mutant-allele specific siRNA against KRAS(G12s), in A549 cells. We observed a dose-dependent anti-proliferative activity against A549 cells treated with exosomes carrying siKRAS(G12s). We observed significant inhibition of A549 tumor xenografts in animals treated with folic acid-functionalized exosomes carrying siKRAS. In summary, milk-derived exosomes represent a viable natural nano-carrier for the delivery of siRNA for therapeutic application against cancer.