Generation and Characterization of Human-Mouse STING Chimeras That Allow DENV Replication in Mouse Cells.
Generation and Characterization of Human-Mouse STING Chimeras That Allow DENV Replication in Mouse Cells.
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本小组首次报道了登革病毒（DENV）在人细胞中直接拮抗环GMP-AMP合成酶(cGAS)/干扰素基因刺激因子（STING）通路的作用，并报道了登革病毒（DENV）非结构蛋白2B (NS2B)-NS3 （NS2B3）蛋白酶复合物与STING相互作用的新发现。我们证实了NS2B与人类STING的跨膜结构域之间的相互作用，以及NS3与人类STING的部分细胞质c端结构域之间的相互作用。我们今天在DENV领域面临的一个重大障碍是缺乏可用的小动物模型，可以在感染的早期事件中有效地概括DENV的发病机制。现有的小鼠模型要么是缺乏干扰素（IFN）受体的免疫功能低下的小鼠，要么是用人类干细胞重组的“人源化”小鼠。然而，这两种方法都未能捕捉到人类发病机制的重要方面，因为它们缺乏关键的先天免疫成分或在免疫细胞发育或维持方面存在缺陷。作为开发DENV免疫小鼠模型的重要一步，我们已经生成了两个嵌合的人-小鼠STING构建体，它们有望在小鼠中保留NS2B3的可切割性和信号传导能力。本文通过构建STING序列缺失突变体来表征人STING与DENV病毒蛋白酶复合物NS2B3的相互作用。研究结果表明，DENV非结构蛋白NS2B与人STING的跨膜结构域相互作用，NS3与人STING的c端环二核苷酸结合结构域相互作用。此外，由于目前还没有理想的具有免疫能力的小鼠模型，可以同时支持DENV的强大复制和概括在人类中观察到的登革热的临床表现，我们表达并表征了两个有希望的人-小鼠嵌合STING构建体，可用于开发相关的转基因小鼠模型，以研究登革热。这两种结构都可以在过表达系统中激活正常的IFN反应，并在感染条件下被裂解。我们相信我们的发现为进一步开发小鼠模型提供了路线图，可以极大地促进DENV的抗病毒发现和疫苗研究。
Our group was the first to describe direct antagonism of the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway by dengue virus (DENV) in human cells, and here, we report new findings on the characterization of the interaction between the DENV nonstructural protein 2B (NS2B)-NS3 (NS2B3) protease complex and STING. We demonstrate interactions between NS2B and the transmembrane domains of human STING and between NS3 and a portion of the cytoplasmic C-terminal domain of human STING. One significant obstacle we face today in the DENV field is the lack of small animal models available that can effectively recapitulate DENV pathogenesis in the early events of infection. The existing mouse models are either immunocompromised mice lacking interferon (IFN) receptors or “humanized” mice reconstituted with human stem cells. However, both approaches fail to capture important aspects of human pathogenesis because they lack critical innate immunity components or have deficiencies in immune cell development or maintenance. As an important step toward developing an immunocompetent mouse model for DENV, we have generated two chimeric human-mouse STING constructs that have promise in retaining both cleavability by NS2B3 and signaling capacity in the mouse. IMPORTANCE This article characterizes the interaction between human STING and DENV viral protease complex NS2B3 by constructing serial deletion mutants of STING. Our findings suggest that DENV nonstructural protein NS2B interacts with the transmembrane domains and NS3 with the C-terminal cyclic dinucleotide binding domain of human STING. Furthermore, as there exists no ideal immunocompetent murine model that can simultaneously support robust DENV replication and recapitulate the clinical manifestation of dengue disease observed in humans, we expressed and characterized two promising human-mouse chimeric STING constructs that can be used for developing a relevant transgenic mouse model to study dengue in the future. Both constructs can activate normal IFN responses in the overexpression system and be cleaved under infection conditions. We believe our findings offer a roadmap to the further development of a murine model that can greatly facilitate antiviral discoveries and vaccine research for DENV.
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影响因子：
5.9
作者：
Chen RE;Diamond MS
通讯作者：
Diamond MS
DOI：
10.3390/v12090979
发表时间：
2020-09-03
期刊：
Viruses
影响因子：
--
作者：
Zhu T;Fernandez-Sesma A
通讯作者：
Fernandez-Sesma A
影响因子：
6.4
作者：
Dalrymple NA;Cimica V;Mackow ER
通讯作者：
Mackow ER
DOI：
10.1016/j.ddmod.2006.03.014
发表时间：
2006-01-01
期刊：
Drug discovery today. Disease models
影响因子：
--
作者：
Bente, Dennis A;Rico-Hesse, Rebeca
通讯作者：
Rico-Hesse, Rebeca
Chikungunya病毒通过降解CGA来拮抗CGAS插入介导的I型干扰素反应。
影响因子：
6.7
作者：
Webb LG;Veloz J;Pintado-Silva J;Zhu T;Rangel MV;Mutetwa T;Zhang L;Bernal-Rubio D;Figueroa D;Carrau L;Fenutria R;Potla U;Reid SP;Yount JS;Stapleford KA;Aguirre S;Fernandez-Sesma A
通讯作者：
Fernandez-Sesma A