A Disintegrin and Metalloproteinase 9 Domain (ADAM9) Is a Major Susceptibility Factor in the Early Stages of Encephalomyocarditis Virus Infection
A Disintegrin and Metalloproteinase 9 Domain (ADAM9) Is a Major Susceptibility Factor in the Early Stages of Encephalomyocarditis Virus Infection
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DOI：
10.1128/mbio.02734-18
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发表时间：
2019-01-01
期刊：
影响因子：
6.4
通讯作者：
Finberg, Robert W.
中科院分区：
文献类型：
--
作者：
Bazzone, Lindsey E.;King, Michael;Finberg, Robert W.
脑心肌炎病毒（EMCV）是一种小核糖核酸病毒，在小鼠和人类细胞中产生裂解性感染。采用全基因组CRISPR-Cas9敲除筛选来寻找EMCV感染所需的宿主因子，我们确定了ADAM 9在EMCV感染中的作用。在多种人类细胞系中，CRISPR介导的ADAM 9缺失使细胞对EMCV感染和细胞死亡具有高度抗性。来自ADAM 9 KO小鼠的原代成纤维细胞也对EMCV感染和细胞死亡具有强烈抗性。相比之下，ADAM 9 KO和WT细胞对其他病毒（包括小核糖核酸病毒科萨基病毒B）的感染同样敏感。当与EMCV孵育时，ADAM 9 KO细胞不能产生病毒子代。然而，通过将病毒RNA直接递送至胞质溶胶而绕过EMCV进入细胞产生了来自ADAM 9 KO细胞的感染性EMCV病毒体，这表明ADAM 9对于EMCV进入后的复制不是必需的。这些研究结果表明，ADAM 9是EMCV感染的早期阶段所必需的，可能是病毒进入或病毒基因组传递到cytosol.Importance病毒性心肌炎是美国死亡的主要原因，导致许多不明原因的死亡在人< 35岁。许多人心肌炎病例都是由肠道病毒引起的。在啮齿动物模型中，脑心肌炎病毒（EMCV）感染引起病毒性心肌炎，但其受体需求尚未完全确定。CRISPR-Cas9筛选可以识别EMCV感染所必需的宿主依赖性因素，并增强我们对病毒感染后关键事件的理解，可能导致预防病毒性心肌炎的新策略。使用CRISPR-Cas9筛选，我们鉴定了去整合素和金属蛋白酶9结构域（ADAM 9）作为人类和小鼠感染中EMCV感染早期阶段所需的主要因素。
Encephalomyocarditis virus (EMCV) is a picornavirus that produces lytic infections in murine and human cells. Employing a genome-wide CRISPR-Cas9 knockout screen to find host factors required for EMCV infection, we identified a role for ADAM9 in EMCV infection. CRISPR-mediated deletion of ADAM9 in multiple human cell lines rendered the cells highly resistant to EMCV infection and cell death. Primary fibroblasts from ADAM9 KO mice were also strongly resistant to EMCV infection and cell death. In contrast, ADAM9 KO and WT cells were equally susceptible to infection with other viruses, including the picornavirus Coxsackie virus B. ADAM9 KO cells failed to produce viral progeny when incubated with EMCV. However, bypassing EMCV entry into cells through delivery of viral RNA directly to the cytosol yielded infectious EMCV virions from ADAM9 KO cells, suggesting that ADAM9 is not required for EMCV replication post-entry. These findings establish that ADAM9 is required for the early stage of EMCV infection, likely for virus entry or viral genome delivery to the cytosol.IMPORTANCE Viral myocarditis is a leading cause of death in the United States, contributing to numerous unexplained deaths in people < 35 years old. Enteroviruses contribute to many cases of human myocarditis. Encephalomyocarditis virus (EMCV) infection causes viral myocarditis in rodent models, but its receptor requirements have not been fully identified. CRISPR-Cas9 screens can identify host dependency factors essential for EMCV infection and enhance our understanding of key events that follow viral infection, potentially leading to new strategies for preventing viral myocarditis. Using a CRISPR-Cas9 screen, we identified a disintegrin and metalloproteinase 9 domain (ADAM9) as a major factor required for the early stages of EMCV infection in both human and murine infection.