Suppression of Adenovirus Replication by Cardiotonic Steroids
Suppression of Adenovirus Replication by Cardiotonic Steroids
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DOI：
10.1128/jvi.01623-16
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发表时间：
2017-02-01
影响因子：
5.4
通讯作者：
Cochrane, Alan
中科院分区：
文献类型：
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作者：
Grosso, Filomena;Stoilov, Peter;Cochrane, Alan
腺病毒依赖于宿主前体RNA剪接机制来表达其完整基因组，这可能使其易受RNA剪接调节剂如地高辛和洋地黄毒苷的影响。这两种药物通过影响基因组复制所需的一个或多个步骤，使四种人腺病毒（HAdV-A31、-B35和-C5以及D种结膜炎分离株）的产量降低至少2至3个对数。立即早期E1 A蛋白水平不受药物影响，但延迟蛋白E4 orf 6和主要晚期衣壳蛋白六邻体的合成受到损害。定量逆转录-PCR（qRT-PCR）分析显示，这两种药物在感染早期改变了E1 A RNA剪接（有利于13 S RNA的产生，而不是12 S RNA），并在病毒复制的晚期部分阻断了从12 S和13 S到9 S RNA的转变。在任一药物存在下，多种晚期病毒蛋白mRNA的表达丢失，与观察到的病毒DNA复制阻断一致。抗病毒作用依赖于药物的持续存在，并且是迅速可逆的。铃兰毒素的衍生物RIDK 34虽然具有更有效的抗病毒活性，但没有显示出改善的选择性指数。这三种药物都在一定程度上降低了代谢活动，但没有细胞死亡的证据。通过在E1 A表达开始之后和基因组复制之前的一个或多个步骤阻断腺病毒复制，地高辛和洋地黄毒苷显示出作为治疗严重腺病毒感染的抗病毒剂的潜力。此外，了解地高辛和洋地黄毒苷抑制腺病毒复制的机制将指导新的抗病毒疗法的发展。重要性尽管人类腺病毒是一种常见的，在某些情况下，危及生命的病原体在人类中，很少有耐受性良好的治疗。在这份报告中，我们证明了两个强心类固醇已经在人类中使用，地高辛和洋地黄毒苷，是多种腺病毒的有效抑制剂。当用HAdV-C5测试时，强心类固醇铃兰毒素的合成衍生物甚至比地高辛和洋地黄毒苷更有效。这些药物改变腺病毒基因表达的级联反应，在早期基因表达启动后起作用，阻断病毒DNA复制和病毒结构蛋白的合成。这些发现证实了一种通过调节宿主细胞过程来治疗腺病毒感染的新方法。
The dependence of adenovirus on the host pre-RNA splicing machinery for expression of its complete genome potentially makes it vulnerable to modulators of RNA splicing, such as digoxin and digitoxin. Both drugs reduced the yields of four human adenoviruses (HAdV-A31, -B35, and -C5 and a species D conjunctivitis isolate) by at least 2 to 3 logs by affecting one or more steps needed for genome replication. Immediate early E1A protein levels are unaffected by the drugs, but synthesis of the delayed protein E4orf6 and the major late capsid protein hexon is compromised. Quantitative reverse transcription-PCR (qRT-PCR) analyses revealed that both drugs altered E1A RNA splicing (favoring the production of 13S over 12S RNA) early in infection and partially blocked the transition from 12S and 13S to 9S RNA at late stages of virus replication. Expression of multiple late viral protein mRNAs was lost in the presence of either drug, consistent with the observed block in viral DNA replication. The antiviral effect was dependent on the continued presence of the drug and was rapidly reversible. RIDK34, a derivative of convallotoxin, although having more potent antiviral activity, did not show an improved selectivity index. All three drugs reduced metabolic activity to some degree without evidence of cell death. By blocking adenovirus replication at one or more steps beyond the onset of E1A expression and prior to genome replication, digoxin and digitoxin show potential as antiviral agents for treatment of serious adenovirus infections. Furthermore, understanding the mechanism(s) by which digoxin and digitoxin inhibit adenovirus replication will guide the development of novel antiviral therapies.IMPORTANCE Despite human adenoviruses being a common and, in some instances, life-threating pathogen in humans, there are few well-tolerated therapies. In this report, we demonstrate that two cardiotonic steroids already in use in humans, digoxin and digitoxin, are potent inhibitors of multiple adenovirus species. A synthetic derivative of the cardiotonic steroid convallotoxin was even more potent than digoxin and digitoxin when tested with HAdV-C5. These drugs alter the cascade of adenovirus gene expression, acting after initiation of early gene expression to block viral DNA replication and synthesis of viral structural proteins. These findings validate a novel approach to treating adenovirus infections through the modulation of host cell processes.