Antiviral Peptides Targeting the Helicase Activity of Enterovirus Nonstructural Protein 2C
Antiviral Peptides Targeting the Helicase Activity of Enterovirus Nonstructural Protein 2C
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DOI：
10.1128/jvi.02324-20
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发表时间：
2021-06-01
影响因子：
5.4
通讯作者：
Zhou, Xi
中科院分区：
文献类型：
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作者：
Fang, Yuan;Wang, Chang;Zhou, Xi
肠道病毒属于短小病毒科肠道病毒属，包括四个人类肠道病毒群(EV-A至D)：人类肠道病毒A71(EV-A71)和柯萨奇病毒A16(CVA16)的流行威胁着全球公共卫生。肠道病毒蛋白2C是肠道病毒中最保守的非结构蛋白，具有在肠道病毒生命周期中起关键作用的RNA解旋酶活性，这使得2C成为开发抗肠病毒药物的一个有吸引力的靶点。在本研究中，我们根据EV-A71 2C的结构设计了一种多肽，命名为2CL。该多肽能有效抑制EV-A71和CVA16编码的2C蛋白的寡聚化，抑制EV-A71和CVA16编码的2C蛋白的RNA解旋酶活性，在细胞内对EV-A71和CVA16具有较强的抗病毒作用。此外，2CL处理对致死性EV-A71攻击具有较强的体内保护作用。此外，以2C解旋酶活性为靶点的抗病毒策略可用于抑制EV-B的复制。无论是2CL还是根据EVB的2CL序列重新设计的多肽B-2CL，都能对两种重要的EV-B，柯萨奇病毒B3和Echo病毒11发挥有效的抗病毒活性。我们的研究结果表明，通过合理设计的多肽来靶向2C的解旋酶活性是一种有效的抗肠道病毒的策略，2CL和B-2CL具有作为广谱抗肠病毒药物的潜在临床开发前景。IMPORTANCE肠道病毒是一大群阳性的单链RNA病毒，包括大量的人类病原体，如肠道病毒A71(EV-A71)、柯萨奇病毒和Echo病毒。然而，目前还没有获得批准的EV抗病毒药物。肠道病毒2C是肠道病毒中最保守的非结构蛋白，含有对病毒生命周期至关重要的RNA解旋酶活性。在此，我们根据EV-A71 2C的结构，设计了一种能有效抑制EV-A71-和柯萨奇病毒A16(CVA16)编码的2C蛋白的RNA解旋酶活性的多肽。此外，该多肽在细胞内对EV-A71和CVA16具有较强的抗病毒作用，在体内对EV-A71致死性攻击具有治疗作用。此外，我们还证明了针对2C解旋酶活性的策略也可以用于其他相关的肠道病毒，包括柯萨奇病毒B3和Echo病毒11。综上所述，我们的发现提供了令人信服的证据，所设计的针对2C解旋酶活性的多肽可能是肠道病毒的广谱抗病毒药物。
Enteroviruses belong to the genus Enterovirus of the family Picomaviridae and include four human enterovirus groups (EV-A to -D): the epidemic of enteroviruses such as human enterovirus A71 (EV-A71) and coxsackievirus A16 (CVA16) is a threat to global public health. Enteroviral protein 2C is the most conserved nonstructural protein among all enteroviruses and possesses RNA helicase activity that plays pivotal roles during enteroviral life cycles, which makes 2C an attractive target for developing antienterovirus drugs. In this study, we designed a peptide, named 2CL, based on the structure of EV-A71 2C. This peptide effectively impaired the oligomerization of EV-A71 2C protein and inhibited the RNA helicase activities of 2C proteins encoded by EV-A71 and CVA16, both of which belong to EV-A, and showed potent antiviral efficacy against EV-A71 and CVA16 in cells. Moreover, the 2CL treatment elicited a strong in vivo protective efficacy against lethal EV-A71 challenge. In addition, the antiviral strategy of targeting the 2C helicase activity can be applied to inhibit the replication of EV-B. Either 2CL or B-2CL, the peptide redesigned based on the 2CL-corresponding sequence of EVBs, could exert effective antiviral activity against two important EV-Bs, coxsackievirus B3 and echovirus 11. Together, our findings demonstrated that targeting the helicase activity of 2C with a rationally designed peptide is an efficient antiviral strategy against enteroviruses, and 2CL and B-2CL show promising clinical potential to be further developed as broad-spectrum antienterovirus drugs.IMPORTANCE Enteroviruses are a large group of positive-sense single-stranded RNA viruses and include numerous human pathogens, such as enterovirus A71 (EV-A71), coxsackieviruses, and echoviruses. However, no approved EV antiviral drugs are available. Enteroviral 2C is the most conserved nonstructural protein among all enteroviruses and contains the RNA helicase activity critical for the viral life cycle. Herein, according to the structure of EV-A71 2C, we designed a peptide that effectively inhibited the RNA helicase activities of EV-A71- and coxsackievirus A16 (CVA16)-encoded 2C proteins. Moreover, this peptide exerted potent antiviral effects against EV-A71 and CVA16 in cells and elicited therapeutic efficacy against lethal EV-A71 challenge in vivo. Furthermore, we demonstrate that the strategy of targeting the 2C helicase activity can be used for other relevant enteroviruses, including coxsackievirus B3 and echovirus 11. In summary, our findings provide compelling evidence that the designed peptides targeting the helicase activity of 2C could be broad-spectrum antivirals for enteroviruses.