乙肝病毒复制体内的RNAi使用激活的慢病毒持续抑制

StephenW

Original Article

Gene Therapy (2015) 22, 163–171; doi:10.1038/gt.2014.94; published online 23 October 2014
Sustained inhibition of hepatitis B virus replication in vivo using RNAi-activating lentiviruses

D Ivacik1, A Ely1, N Ferry2 and P Arbuthnot1

    1Antiviral Gene Therapy Research Unit, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, Gauteng, South Africa
    2Agence Nationale de Sécurité du Médicament et des produits de Santé, Saint Denis, France

Correspondence: Dr P Arbuthnot, Antiviral Gene Therapy Research Unit, School of Pathology, Health Sciences Faculty, University of the Witwatersrand, 7 York Road, Parktown, Johannesburg 2193, Gauteng, South Africa. E-mail: [email protected]

Received 17 July 2014; Revised 26 August 2014; Accepted 17 September 2014
Advance online publication 23 October 2014
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Abstract

Chronic infection with hepatitis B virus (HBV) puts individuals at high risk for complicating cirrhosis and liver cancer, but available treatment to counter the virus rarely eliminates infection. Although harnessing RNA interference (RNAi) to silence HBV genes has shown the potential, achieving efficient and durable silencing of viral genes remains an important goal. Here we report on the propagation of lentiviral vectors (LVs) that successfully deliver HBV-targeting RNAi activators to liver cells. Mono- and tricistronic artificial primary microRNAs (pri-miRs) derived from pri-miR-31, placed under transcriptional control of the liver-specific modified murine transthyretin (mTTR) promoter, caused efficient inhibition of HBV replication markers. The tricistronic cassette was capable of silencing a mutant viral target and the effects were observed without disrupting the function of an endogenous miR (miR-16). The mTTR promoter stably expressed a reporter transgene in mouse livers over a study period of 1 year. Good silencing of HBV genes, without evidence of toxicity, was demonstrated following intravenous injection of LVs into neonatal HBV transgenic mice. Collectively, these data indicate that LVs may achieve sustained inhibition of HBV replication that is appealing for their therapeutic use.

StephenW

原来的文章

基因治疗（2015）22，163-171; DOI：10.1038/ gt.2014.94;网上公布的23月到2014年
乙肝病毒复制体内的RNAi使用激活的慢病毒持续抑制

ðIvacik1，A Ely1，N Ferry2和P Arbuthnot1

    1Antiviral基因治疗研究单位，病理学院，健康科学学院，威特沃特斯兰德大学，约翰内斯堡，豪登省，南非
    2Agence国民安全局杜药剂等DES PRODUITS德桑特，圣但尼，法国

函授：博士P亚毕诺，抗病毒基因治疗研究单位，病理学院，健康科学学院，威特沃特斯兰德大学，约克路7号，Parktown，约翰内斯堡2193，豪登省，南非。电子邮件：[email protected]

收到2014年7月17日;修订后的2014年8月26日;接受2014年9月17日
推进网上公布2014年10月23日
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摘要

慢性感染乙型肝炎病毒（HBV）使个人在高风险的复杂性肝硬化和肝癌，但可用的治疗，以对抗病毒很少消除感染。虽然利用RNA干扰（RNAi）沉默的HBV基因已示出的电势，从而实现病毒基因的高效和耐用沉默仍是一个重要的目标。在这里，我们报告慢病毒载体（LVS），成功地实现HBV靶向RNA干扰激活肝细胞的繁殖。单从PRI-的miR-31衍生的三顺反子人工小学的microRNA（PRI-大鹏），在肝特异性修饰的小鼠甲状腺素（MTTR）启动子的转录控制之下，所引起的HBV复制指标有效抑制。该三顺反子盒是能够沉默突变的病毒靶和不中断的内源性的miR（的miR-16）的功能被观察到的影响。该MTTR促进稳定表达的小鼠肝脏记者转基因超过1年的研究期间。好沉默HBV基因，而没有毒性的证据，证明在静脉注射的LV到新生的HBV转基因小鼠。总的来说，这些数据表明，可能的LV实现HBV复制所呼吁它们的治疗用途的持续抑制。