RNAi-mediated gene knockdown and anti-angiogenic therapy of RCCs using a cyclic RGD-modified liposomal-siRNA system
RNAi-mediated gene knockdown and anti-angiogenic therapy of RCCs using a cyclic RGD-modified liposomal-siRNA system
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DOI：
10.1016/j.jconrel.2013.10.003
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发表时间：
2014-01-10
影响因子：
10.8
通讯作者：
Harashima, Hideyoshi
中科院分区：
文献类型：
--
作者：
Sakurai, Yu;Hatakeyama, Hiroto;Harashima, Hideyoshi
血管生成是与肿瘤生长和发展相关的关键过程之一，因此是癌症治疗的主要靶点。虽然肿瘤内皮细胞（TEC）在病理性血管生成中起着关键作用，但当TEC中的基因表达被抑制时，研究新血管的表型变化是一项困难的任务。小干扰RNA（siRNA）由于其使感兴趣的基因沉默的能力而代表潜在的试剂。我们先前开发了一种用于体内siRNA递送至癌细胞的系统，该系统涉及脂质体递送系统，一种含有独特的pH敏感性阳离子脂质YSK 05（YSK-MEND）的MEND。在本研究中，我们报告了一个系统的发展，允许通过结合YSK-MEND和一个配体，是特定的TEC的siRNA到TEC的交付。环（Arg-Gly-Asp-D-Phe-Lys）（cRGD）是α（V）β（3）整联蛋白的众所周知的配体，α（V）β（3）整联蛋白在TEC中以高水平选择性表达。我们将cRGD掺入YSK-MEND（RGD-MEND）中以实现TEC中的有效基因沉默。定量RT-PCR和5'端cDNA快速扩增PCR结果表明，静脉注射RGD-MEND 4.0mg/kg可显著降低TEC中RNAi介导的基因表达，但对其他器官的内皮细胞无明显影响。最后，我们评估了RGD-MEND封装siRNA对血管内皮生长因子受体2的治疗效力。隔日连续三次注射RGD-MEND后，观察到肿瘤生长显著延迟。总之，RGD-MEND代表了一种新的方法来表征TEC和我们在抗血管生成治疗。(C)2013爱思唯尔有限公司版权所有。
Angiogenesis is one of crucial processes associated with tumor growth and development, and consequently a prime target for cancer therapy. Although tumor endothelial cells (TECs) play a key role in pathological angiogenesis, investigating phenotypical changes in neovessels when a gene expression in TEC is suppressed is a difficult task. Small interfering RNA (siRNA) represents a potential agent due to its ability to silence a gene of interest. We previously developed a system for in vivo siRNA delivery to cancer cells that involves a liposomal-delivery system, a MEND that contains a unique pH-sensitive cationic lipid, YSK05 (YSK-MEND). In the present study, we report on the development of a system that permits the delivery of siRNA to TECs by combining the YSK-MEND and a ligand that is specific to TECs. Cyclo(Arg-Gly-Asp-D-Phe-Lys) (cRGD) is a well-known ligand to alpha(V)beta(3) integrin, which is selectively expressed at high levels in TECs. We incorporated cRGD into the YSK-MEND (RGD-MEND) to achieve an efficient gene silencing in TECs. Quantitative RT-PCR and the 5' rapid amplification of cDNA ends PCR indicated that the intravenous injection of RGD-MEND at a dose of 4.0 mg/kg induced a significant RNAi-mediated gene reduction in TEC but not in endothelial cells of other organs. Finally, we evaluated the therapeutic potency of the RGD-MEND encapsulating siRNA against vascular endothelial growth factor receptor 2. A substantial delay in tumor growth was observed after three sequential RGD-MEND injections on alternate days. In conclusion, the RGD-MEND represents a new approach for the characterization of TECs and for us in anti-angiogenic therapy. (C) 2013 Elsevier B.V. All rights reserved.