CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages.
CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages.
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DOI：
10.1002/jlb.3mia0917-352r
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发表时间：
2018-02-13
影响因子：
5.5
通讯作者：
Wallet MA
中科院分区：
文献类型：
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作者：
Taylor JP;Cash MN;Santostefano KE;Nakanishi M;Terada N;Wallet MA
IFN刺激基因泛素特异性蛋白酶18 （USP18）编码一种蛋白，该蛋白通过硬脂质抑制JAK1向IFN α受体2亚基（IFNAR2）募集，负调控T1 IFN信号。在这里，我们证明了USP18的表达是由HIV‐1以T1依赖IFN的方式诱导的。在诱导多能干细胞（iPSC）衍生的巨噬细胞模型中，通过聚集规律间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9 （Cas9）基因编辑去除USP18的实验结果显著限制了HIV - 1的复制。在USP18缺失的情况下，巨噬细胞对T1 IFN刺激的反应性增加，STAT1和STAT2的磷酸化延长，IFN刺激基因的表达增加，这是抗病毒反应的关键。有趣的是，HIV‐1需要一些通过T1 IFN受体的信号传导才能有效地复制，因为抑制T1 IFN活性的中和抗体可以降低单核细胞源性巨噬细胞中HIV‐1的复制率。HIV‐1诱导USP18将IFN反应调节到最佳水平，允许从HIV‐1 LTR启动子高效转录，同时最小化T1 IFN诱导的抗病毒反应，否则将限制病毒复制和传播。最后，iPSC和CRISPR/Cas9基因靶向为研究调节先天免疫反应的宿主因子提供了一个强大的工具。USP18通过负调控T1 - IFN信号诱导的抗病毒基因表达，促进HIV - 1复制。
The IFN‐stimulated gene ubiquitin‐specific proteinase 18 (USP18) encodes a protein that negatively regulates T1 IFN signaling via stearic inhibition of JAK1 recruitment to the IFN‐α receptor 2 subunit (IFNAR2). Here, we demonstrate that USP18 expression is induced by HIV‐1 in a T1 IFN‐dependent manner. Experimental depletion of USP18 by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9) gene editing results in a significant restriction of HIV‐1 replication in an induced pluripotent stem cell (iPSC)‐derived macrophage model. In the absence of USP18, macrophages have increased responsiveness to stimulation with T1 IFNs with prolonged phosphorylation of STAT1 and STAT2 and increased expression of IFN‐stimulated genes that are key for antiviral responses. Interestingly, HIV‐1 requires some signaling through the T1 IFN receptor to replicate efficiently because a neutralizing antibody that inhibits T1 IFN activity reduces HIV‐1 replication rate in monocyte‐derived macrophages. USP18 induction by HIV‐1 tunes the IFN response to optimal levels allowing for efficient transcription from the HIV‐1 LTR promoter while minimizing the T1 IFN‐induced antiviral response that would otherwise restrict viral replication and spread. Finally, iPSC and CRISPR/Cas9 gene targeting offer a powerful tool to study host factors that regulate innate immune responses. USP18 promotes HIV‐1 replication by negatively regulating expression of antiviral genes induced by T1 IFN signaling.