Gene Silencing via RNAi and siRNA Quantification in Tumor Tissue Using MEND, a Liposomal siRNA Delivery System
Gene Silencing via RNAi and siRNA Quantification in Tumor Tissue Using MEND, a Liposomal siRNA Delivery System
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DOI：
10.1038/mt.2013.57
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发表时间：
2013-06-01
影响因子：
12.4
通讯作者：
Harashima, Hideyoshi
中科院分区：
文献类型：
--
作者：
Sakurai, Yu;Hatakeyama, Hiroto;Harashima, Hideyoshi
预计小干扰RNA（siRNA）可用作癌症药物，但临床开发需要高效的siRNA递送系统。为了解决这个问题，我们开发了一种脂质体siRNA载体，一种多功能的介孔型纳米器件（MEND）。我们先前报道了由pH敏感的阳离子脂质YSK 05组成的MEND在体外和肿瘤组织中通过瘤内注射显示出显著的敲低。在这里，我们报告的发展，在体内siRNA传递系统，通过全身注射和静脉注射给药的siRNA分子在肿瘤组织中的药代动力学分析。通过茎环引物定量逆转录酶PCR（qRT-PCR）方法定量siRNA的肿瘤递送。YSK-MEND的PEG化导致siRNA在肿瘤组织中的积累从0.0079%ID/g肿瘤增加至1.9%ID/g肿瘤。施用MEND（3 mg siRNA/kg体重）显示靶基因mRNA和蛋白质减少约50%。此外，我们还通过5' RACE-PCR方法验证了RNA干扰的诱导。本文报道的集体结果表明，开发了一种siRNA载体，其可以通过改善的siRNA生物利用度将siRNA递送至肿瘤组织中的靶细胞。
Small interfering RNA (siRNA) would be predicted to function as a cancer drug, but an efficient siRNA delivery system is required for clinical development. To address this issue, we developed a liposomal siRNA carrier, a multifunctional envelope-type nanodevice (MEND). We previously reported that a MEND composed of a pH-sensitive cationic lipid, YSK05, showed significant knockdown in both in vitro and in tumor tissue by intratumoral injection. Here, we report on the development of an in vivo siRNA delivery system that is delivered by systemic injection and an analysis of the pharmacokinetics of an intravenously administered siRNA molecule in tumor tissue. Tumor delivery of siRNA was quantified by means of stem-loop primer quantitative reverse transcriptase PCR (qRT-PCR) method. PEGylation of the YSK-MEND results in the increase in the accumulation of siRNA in tumor tissue from 0.0079% ID/g tumor to 1.9% ID/g tumor. The Administration of the MEND (3 mg siRNA/kg body weight) showed about a 50% reduction in the target gene mRNA and protein. Moreover, we verified the induction of RNA interference by 5' RACE-PCR method. The collective results reported here indicate that an siRNA carrier was developed that can deliver siRNA to a target cell in tumor tissue through an improved siRNA bioavailability.