Ashbrook Alison W, Lentscher Anthony J, Zamora Paula F, Silva Laurie A, May Nicholas A, Bauer Joshua A, Morrison Thomas E, Dermody Terence S
Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University School of Medicine, Nashville, Tennessee, USA Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
mBio. 2016 May 24;7(3):e00693-16. doi: 10.1128/mBio.00693-16.
Chikungunya virus (CHIKV) is a reemerging alphavirus that has caused epidemics of fever, arthralgia, and rash worldwide. There are currently no licensed vaccines or antiviral therapies available for the prevention or treatment of CHIKV disease. We conducted a high-throughput, chemical compound screen that identified digoxin, a cardiac glycoside that blocks the sodium-potassium ATPase, as a potent inhibitor of CHIKV infection. Treatment of human cells with digoxin or a related cardiac glycoside, ouabain, resulted in a dose-dependent decrease in infection by CHIKV. Inhibition by digoxin was cell type-specific, as digoxin treatment of either murine or mosquito cells did not diminish CHIKV infection. Digoxin displayed antiviral activity against other alphaviruses, including Ross River virus and Sindbis virus, as well as mammalian reovirus and vesicular stomatitis virus. The digoxin-mediated block to CHIKV and reovirus infection occurred at one or more postentry steps, as digoxin inhibition was not bypassed by fusion of CHIKV at the plasma membrane or infection with cell surface-penetrating reovirus entry intermediates. Selection of digoxin-resistant CHIKV variants identified multiple mutations in the nonstructural proteins required for replication complex formation and synthesis of viral RNA. These data suggest a role for the sodium-potassium ATPase in promoting postentry steps of CHIKV replication and provide rationale for modulation of this pathway as a broad-spectrum antiviral strategy.
Mitigation of disease induced by globally spreading, mosquito-borne arthritogenic alphaviruses requires the development of new antiviral strategies. High-throughput screening of clinically tested compounds provides a rapid means to identify undiscovered, antiviral functions for well-characterized therapeutics and illuminate host pathways required for viral infection. Our study describes the potent inhibition of Chikungunya virus and related alphaviruses by the cardiac glycoside digoxin and demonstrates a function for the sodium-potassium ATPase in Chikungunya virus infection.
基孔肯雅病毒（CHIKV）是一种重新出现的甲病毒，已在全球范围内引发发热、关节痛和皮疹的流行。目前尚无用于预防或治疗基孔肯雅病毒病的许可疫苗或抗病毒疗法。我们进行了一项高通量化合物筛选，确定了地高辛（一种阻断钠钾ATP酶的强心苷）是基孔肯雅病毒感染的有效抑制剂。用地高辛或相关强心苷哇巴因处理人细胞，导致基孔肯雅病毒感染呈剂量依赖性下降。地高辛的抑制作用具有细胞类型特异性，因为用地高辛处理小鼠或蚊子细胞不会减少基孔肯雅病毒感染。地高辛对其他甲病毒具有抗病毒活性，包括罗斯河病毒和辛德毕斯病毒，以及哺乳动物呼肠孤病毒和水泡性口炎病毒。地高辛介导的对基孔肯雅病毒和呼肠孤病毒感染的阻断发生在一个或多个进入后步骤，因为地高辛的抑制作用不会因基孔肯雅病毒在质膜处的融合或用细胞表面穿透性呼肠孤病毒进入中间体感染而被绕过。选择抗地高辛的基孔肯雅病毒变体鉴定出在复制复合物形成和病毒RNA合成所需的非结构蛋白中有多个突变。这些数据表明钠钾ATP酶在促进基孔肯雅病毒复制的进入后步骤中起作用，并为将该途径作为一种广谱抗病毒策略进行调节提供了理论依据。
减轻由全球传播的、蚊媒致关节炎甲病毒引起的疾病需要开发新的抗病毒策略。对经过临床测试的化合物进行高通量筛选提供了一种快速方法，以确定特征明确的治疗药物未被发现的抗病毒功能，并阐明病毒感染所需的宿主途径。我们的研究描述了强心苷地高辛对基孔肯雅病毒和相关甲病毒的有效抑制，并证明了钠钾ATP酶在基孔肯雅病毒感染中的作用。
