A yellow fever virus NS4B inhibitor not only suppresses viral replication, but also enhances the virus activation of RIG-I-like receptor-mediated innate immune response.
A yellow fever virus NS4B inhibitor not only suppresses viral replication, but also enhances the virus activation of RIG-I-like receptor-mediated innate immune response.
复制标题
DOI：
10.1371/journal.ppat.1010271
复制
发表时间：
2022-01
期刊：
影响因子：
6.7
通讯作者：
Chang J
中科院分区：
文献类型：
--
作者：
Gao Z;Zhang X;Zhang L;Wu S;Ma J;Wang F;Zhou Y;Dai X;Bullitt E;Du Y;Guo JT;Chang J
黄病毒感染细胞诱导内质网（ER）膜的大量重排，形成病毒复制细胞器（RO），将病毒RNA复制中间体与细胞质RNA传感器分离。在其他病毒非结构（NS）蛋白中，现有证据表明NS 4 B（一种具有多个跨膜结构域的ER膜蛋白）在RO的形成和先天性免疫应答的逃避中起着重要作用。我们先前报道了一种苯二氮卓类化合物BDAA，它特异性地抑制黄热病毒（YFV）在培养细胞和仓鼠体内的复制，其耐药突变定位于NS 4 B蛋白的P219。在接下来的机制研究中，我们发现BDAA特异性增强YFV诱导的炎症细胞因子反应，与诱导RO的显着结构改变和病毒感染细胞中双链RNA（dsRNA）的暴露相关。有趣的是，在YFV感染的细胞中BDAA增强的细胞因子应答在RIG-I或MAD 5敲除细胞中减弱，并且在MAVS敲除细胞中完全消除。然而，BDAA在亲本细胞和RIG-I、MDA 5或MAVS缺陷的细胞中以相似的程度抑制YFV复制。因此，这些结果提供了多条生物学证据来支持BDAA与NS 4 B的相互作用可能损害YFV R 0的完整性的模型，这不仅抑制病毒RNA复制，而且还促进病毒RNA从R 0的释放，从而激活RIG-I和MDA 5。虽然BDAA的天然免疫增强活性不是其在培养细胞中的抗病毒活性所必需的，但其双重抗病毒机制在迄今为止报道的所有抗病毒药物中是独特的，值得在未来的动物模型中进一步研究。由黄热病病毒（YFV）感染引起的黄热病（YF）的出现和重新出现已经在以前的非流行地区以及流行地区构成了全球公共卫生威胁。大约30%的死亡率使疫情特别具有破坏性。除了疫苗接种和蚊虫控制外，抗病毒药物也是防治黄热病爆发的重要手段。然而，目前只有两种为治疗其他RNA病毒感染而开发的核苷酸类似物药物被重新用于治疗YF，临床疗效不确定。BDAA是我们实验室发现的一种有效的YFV特异性抗病毒剂。本文报道的工作进一步证明BDAA与YFV NS 4 B蛋白的相互作用可能损害病毒RNA复制细胞器的完整性，这不仅抑制病毒RNA复制，而且导致病毒RNA泄漏到细胞质中以激活RIG-I样RNA受体并增强先天性抗病毒免疫应答。BDAA前所未有的抗病毒机制突出了NS 4 B蛋白在病毒RNA复制和逃避宿主细胞先天免疫中的重要作用。
Flavivirus infection of cells induces massive rearrangements of the endoplasmic reticulum (ER) membrane to form viral replication organelles (ROs) which segregates viral RNA replication intermediates from the cytoplasmic RNA sensors. Among other viral nonstructural (NS) proteins, available evidence suggests for a prominent role of NS4B, an ER membrane protein with multiple transmembrane domains, in the formation of ROs and the evasion of the innate immune response. We previously reported a benzodiazepine compound, BDAA, which specifically inhibited yellow fever virus (YFV) replication in cultured cells and in vivo in hamsters, with resistant mutation mapped to P219 of NS4B protein. In the following mechanistic studies, we found that BDAA specifically enhances YFV induced inflammatory cytokine response in association with the induction of dramatic structural alteration of ROs and exposure of double-stranded RNA (dsRNA) in virus-infected cells. Interestingly, the BDAA-enhanced cytokine response in YFV-infected cells is attenuated in RIG-I or MAD5 knockout cells and completely abolished in MAVS knockout cells. However, BDAA inhibited YFV replication at a similar extent in the parent cells and cells deficient of RIG-I, MDA5 or MAVS. These results thus provided multiple lines of biological evidence to support a model that BDAA interaction with NS4B may impair the integrity of YFV ROs, which not only inhibits viral RNA replication, but also promotes the release of viral RNA from ROs, which consequentially activates RIG-I and MDA5. Although the innate immune enhancement activity of BDAA is not required for its antiviral activity in cultured cells, its dual antiviral mechanism is unique among all the reported antiviral agents thus far and warrants further investigation in animal models in future. Emergence and re-emergence of yellow fever (YF) caused by the yellow fever virus (YFV) infection have posed a global public health threat in previously non-epidemic as well as endemic regions. The approximately 30% of mortality rate makes the outbreaks particularly devastating. In addition to the vaccination campaign and mosquito controls, antiviral drugs are important components in the toolbox for combating YF outbreaks. However, only two nucleotide analogue drugs developed for the treatment of other RNA virus infections are currently repurposed for the treatment of YF with uncertain clinical efficacy. BDAA is a benzodiazepine compound discovered as a potent YFV-specific antiviral agent in our laboratory. The work reported herein further demonstrates that BDAA interaction with the YFV NS4B protein may impair the integrity of viral RNA replication organelles, which not only inhibits viral RNA replication, but also results in the leakage of viral RNA into the cytoplasm to activate RIG-I-like RNA receptors and enhances the innate antiviral immune response. The unprecedented antiviral mechanism of BDAA highlights the essential role of the NS4B protein in viral RNA replication and the evasion of host cellular innate immunity.
登录
查看更多内容
影响因子：
2
作者：
Bifani AM;Ong EZ;de Alwis R
通讯作者：
de Alwis R
影响因子：
5.4
作者：
Guo, Fang;Wu, Shuo;Chang, Jinhong
通讯作者：
Chang, Jinhong
影响因子：
16.6
作者：
Douam F;Ziegler CGK;Hrebikova G;Fant B;Leach R;Parsons L;Wang W;Gaska JM;Winer BY;Heller B;Shalek AK;Ploss A
通讯作者：
Ploss A
影响因子：
3.8
作者：
Du, Xiaoting;Zhang, Yang;Yi, Zhigang
通讯作者：
Yi, Zhigang
影响因子：
5.4
作者：
Errett, John S.;Suthar, Mehul S.;Gale, Michael, Jr.
通讯作者：
Gale, Michael, Jr.