Hepatic cell-to-cell transmission of small silencing RNA can extend the therapeutic reach of RNA interference (RNAi)
Hepatic cell-to-cell transmission of small silencing RNA can extend the therapeutic reach of RNA interference (RNAi)
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DOI：
10.1136/gutjnl-2011-300449
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发表时间：
2012-09-01
期刊：
影响因子：
24.5
通讯作者：
van der Laan, Luc J. W.
中科院分区：
文献类型：
--
作者：
Pan, Qiuwei;Ramakrishnaiah, Vedashree;van der Laan, Luc J. W.
背景/目的RNA干扰(RNAi)是一种由小干扰RNA(SiRNA)触发的序列特异性基因沉默技术，是治疗包括丙型肝炎病毒(HCV)感染在内的多种肝病的新途径。在植物和无脊椎动物中，RNAi提供了一种重要的细胞防御病毒病原体的机制，并依赖于siRNA向邻近细胞的传播。为探讨载体介导的RNAi能否在体外和小鼠肝细胞间转移，以及这种交换是否能延长RNAi对丙型肝炎病毒感染的治疗效果。方法利用人肝细胞系和转导siRNA表达载体的原代B淋巴细胞，通过沉默丙型肝炎病毒复制和病毒进入受体CD81的表达，在培养条件下研究RNAi的传递。在NOD/SCID小鼠体内研究肝细胞间的传递。结果人和小鼠肝细胞以及原代人B细胞具有交换小RNA的能力，包括细胞内源性microRNA，并能递送针对丙型肝炎病毒或CD81的siRNA。RNAi的传递在很大程度上不依赖于细胞接触，部分由外切体介导。在NOD/SCID小鼠体内观察到RNAi传递的证据是产生CD81 siRNA的人肝癌细胞，导致CD81在小鼠肝细胞中的表达受到抑制。结论人和小鼠肝细胞都可以交换小分子沉默RNA，部分是通过外切体的穿梭来实现的。SiRNA的传播潜在地扩大了基于RNAi的治疗丙型肝炎和其他肝病的范围。
Background/aims RNA interference (RNAi), a sequence-specific gene silencing technology triggered by small interfering RNA (siRNA), represents promising new avenues for treatment of various liver diseases including hepatitis C virus (HCV) infection. In plants and invertebrates, RNAi provides an important mechanism of cellular defence against viral pathogens and is dependent on the spread of siRNA to neighbouring cells. A study was undertaken to investigate whether vector-delivered RNAi can transfer between hepatic cells in vitro and in mice, and whether this exchange could extend the therapeutic effect of RNAi against HCV infection.Methods Transmission of RNAi was investigated in culture by assessing silencing of HCV replication and expression of viral entry receptor CD81 using a human hepatic cell line and primary B lymphocytes transduced with siRNA-expressing vectors. In vivo transmission between hepatic cells was investigated in NOD/SCID mice. Involvement of exosomes was demonstrated by purification, uptake and mass spectrometric analysis.Results Human and mouse liver cells, as well as primary human B cells, were found to have the ability to exchange small RNAs, including cellular endogenous microRNA and delivered siRNA targeting HCV or CD81. The transmission of RNAi was largely independent of cell contact and partially mediated by exosomes. Evidence of RNAi transmission in vivo was observed in NOD/SCID mice engrafted with human hepatoma cells producing CD81 siRNA, causing suppression of CD81 expression in mouse hepatocytes.Conclusion Both human and mouse hepatic cells exchange small silencing RNAs, partially mediated by shuttling of exosomes. Transmission of siRNA potentially extends the therapeutic reach of RNAi-based therapies against HCV as well as other liver diseases.