The retrovirus human immunodeficiency virus-1 (HIV-1) is the causative agent of AIDS. Although treatment of HIV/AIDS with antiretroviral therapy provides suppression of viremia, latent reservoirs of integrated proviruses preclude cure by current antiviral treatments. Understanding the mechanisms of host–viral interactions may elucidate new treatment strategies. Here, we performed a CRISPR/Cas9 transcriptional activation screen using a high-complexity, genome-wide sgRNA library to identify cellular factors that inhibit HIV-1 infection of human CD4+ T cells. MT4 cells were transduced with a CRISPR/Cas9 sgRNA library and infected with nef-deficient HIV-1NL4-3 expressing ganciclovir-sensitive thymidine kinase, thus enabling selection of HIV-1-resistant cells for analysis of enriched sgRNAs. After validation of screen hits, multiple host factors essential for HIV-1 infection were identified, including SET (SET nuclear proto-oncogene) and ANP32A (acidic nuclear phosphoprotein 32A, PP32A), which together form a histone acetylase inhibitor complex. Using multiple human cell lines and peripheral blood mononuclear cells (PBMCs) from healthy donors and HIV-1-infected individuals, we demonstrate that SET depletion increased HIV-1 infectivity by augmenting DNA integration without significantly changing sites of integration. Conversely, SET overexpression decreased HIV-1 integration and infectivity. SET protein expression was significantly reduced in PBMCs from HIV-1-infected individuals and was downregulated by HIV-1 infection of healthy donor cells in vitro. Notably, HIV-1-induced downregulation of SET could be alleviated by inhibition of the protease granzyme A. Altogether, we have identified cellular inhibitors of HIV-1 infection on a genome-wide scale, which affords new insight into host–virus interactions and may provide new strategies for HIV-1 treatment.

中文翻译：

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全基因组 CRISPR/Cas9 转录激活筛选确定组蛋白乙酰转移酶抑制剂复合物作为 HIV-1 整合的调节剂

逆转录病毒人类免疫缺陷病毒 1 (HIV-1) 是艾滋病的病原体。尽管用抗逆转录病毒疗法治疗 HIV/AIDS 可抑制病毒血症，但整合原病毒的潜在储库无法通过当前的抗病毒疗法治愈。了解宿主-病毒相互作用的机制可能会阐明新的治疗策略。在这里，我们使用高度复杂的全基因组 sgRNA 文库进行了 CRISPR/Cas9 转录激活筛选，以确定抑制 HIV-1 感染人类 CD4+ T 细胞的细胞因子。MT4 细胞用 CRISPR/Cas9 sgRNA 文库转导，并感染表达更昔洛韦敏感胸苷激酶的 nef 缺陷型 HIV-1NL4-3，从而能够选择 HIV-1 抗性细胞来分析富集的 sgRNA。屏幕命中验证后，确定了 HIV-1 感染所必需的多种宿主因子，包括 SET（SET 核原癌基因）和 ANP32A（酸性核磷蛋白 32A，PP32A），它们共同形成组蛋白乙酰化酶抑制剂复合物。我们使用来自健康供体和 HIV-1 感染者的多个人类细胞系和外周血单核细胞 (PBMC)，证明 SET 耗竭通过增强 DNA 整合而增加了 HIV-1 传染性，而没有显着改变整合位点。相反，SET 过表达降低了 HIV-1 整合和传染性。SET 蛋白表达在来自 HIV-1 感染个体的 PBMC 中显着降低，并且在体外被健康供体细胞的 HIV-1 感染下调。值得注意的是，抑制蛋白酶颗粒酶 A 可以减轻 HIV-1 诱导的 SET 下调。