The Long Noncoding RNA HEAL Regulates HIV-1 Replication through Epigenetic Regulation of the HIV-1 Promoter
The Long Noncoding RNA HEAL Regulates HIV-1 Replication through Epigenetic Regulation of the HIV-1 Promoter
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DOI：
10.1128/mbio.02016-19
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发表时间：
2019-09-01
期刊：
影响因子：
6.4
通讯作者：
Rana, Tariq M.
中科院分区：
文献类型：
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作者：
Chao, Ti-Chun;Zhang, Qiong;Rana, Tariq M.
寻找HIV-1/AIDS治愈方法的一个主要挑战是难以识别和根除具有复制能力的前病毒的持久储存库。长链非编码RNA（lncRNA，>200个核苷酸）在病理生理学中发挥着重要作用。在这里，我们报告的第一个全基因组的lncRNA在HIV-1感染的主要单核细胞衍生的巨噬细胞（MDM）的表达分析。我们鉴定了一种lncRNA，我们将其命名为HIV-1增强的lncRNA（HEAL），它被MDM，小胶质细胞和T淋巴细胞的HIV-1感染上调。HIV-1感染者的外周血单个核细胞显示HEAL水平升高。重要的是，HEAL是多种HIV-1毒株的广泛增强子，因为HEAL的耗尽抑制了X4、R5和双嗜性HIV复制，并且这种抑制被HEAL过表达所挽救。HEAL与RNA结合蛋白FUS形成复合物，通过至少两种机制促进HIV复制：（i）HEAL-FUS复合物结合HIV启动子并增强组蛋白乙酰转移酶p300的募集，所述组蛋白乙酰转移酶p300通过增加HIV启动子上的组蛋白H3 K27乙酰化和P-TEFb富集来正向调节HIV转录，和（ii）HEAL-FUS复合物在细胞周期蛋白依赖性激酶2基因CDK 2的启动子处富集，以增强CDK 2表达。值得注意的是，分别由RNA干扰（RNAi）和CRISPR-Cas9介导的HEAL敲低和敲除在体外停止叠氮胸苷治疗后防止T细胞和小胶质细胞中的HIV-1复发。我们的研究结果表明，沉默的HEAL或干扰的HEAL-FUS核糖核蛋白复合物可以提供一种新的表观遗传沉默策略，以消除病毒水库和效果治愈HIV-1/AIDS。重要的是，尽管我们增加了对lncRNA的功能的理解，他们的潜力，开发HIV/AIDS治愈策略仍然未被探索。对HIV-1感染的原代单核细胞衍生的巨噬细胞（MDM）中的lncRNA进行了全基因组分析，并鉴定了1，145种差异表达的lncRNA。一种名为HIV-1增强型lncRNA（HEAL）的lncRNA被HIV-1感染上调，并促进T细胞和巨噬细胞中的HIV复制。HEAL与RNA结合蛋白FUS形成复合物，以增强转录辅激活因子p300向HIV启动子的募集。此外，HEAL敲低和敲除可防止在停止叠氮胸苷治疗后T细胞和小胶质细胞中的HIV-1复发，这表明HEAL是治愈HIV-1/AIDS的潜在治疗靶点。
A major challenge in finding a cure for HIV-1/AIDS is the difficulty in identifying and eradicating persistent reservoirs of replication-competent provirus. Long noncoding RNAs (lncRNAs, >200 nucleotides) are increasingly recognized to play important roles in pathophysiology. Here, we report the first genome-wide expression analysis of lncRNAs in HIV-1-infected primary monocyte-derived macrophages (MDMs). We identified an lncRNA, which we named HIV-1-enhanced lncRNA (HEAL), that is upregulated by HIV-1 infection of MDMs, microglia, and T lymphocytes. Peripheral blood mononuclear cells of HIV-1-infected individuals show elevated levels of HEAL. Importantly, HEAL is a broad enhancer of multiple HIV-1 strains because depletion of HEAL inhibited X4, R5, and dual-tropic HIV replications and the inhibition was rescued by HEAL overexpression. HEAL forms a complex with the RNA-binding protein FUS, which facilitates HIV replication through at least two mechanisms: (i) HEAL-FUS complex binds the HIV promoter and enhances recruitment of the histone acetyltransferase p300, which positively regulates HIV transcription by increasing histone H3K27 acetylation and P-TEFb enrichment on the HIV promoter, and (ii) HEAL-FUS complex is enriched at the promoter of the cyclin-dependent kinase 2 gene, CDK2, to enhance CDK2 expression. Notably, HEAL knockdown and knockout mediated by RNA interference (RNAi) and CRISPR-Cas9, respectively, prevent HIV-1 recrudescence in T cells and microglia upon cessation of azidothymidine treatment in vitro. Our results suggest that silencing of HEAL or perturbation of the HEAL-FUS ribonucleoprotein complex could provide a new epigenetic silencing strategy to eradicate viral reservoirs and effect a cure for HIV-1/AIDS.IMPORTANCE Despite our increased understanding of the functions of lncRNAs, their potential to develop HIV/AIDS cure strategies remains unexplored. A genome-wide analysis of lncRNAs in HIV-1-infected primary monocyte-derived macrophages (MDMs) was performed, and 1,145 differentially expressed lncRNAs were identified. An lncRNA named HIV-1-enhanced lncRNA (HEAL) is upregulated by HIV-1 infection and promotes HIV replication in T cells and macrophages. HEAL forms a complex with the RNA-binding protein FUS to enhance transcriptional coactivator p300 recruitment to the HIV promoter. Furthermore, HEAL knockdown and knockout prevent HIV-1 recrudescence in T cells and microglia upon cessation of azidothymidine treatment, suggesting HEAL as a potential therapeutic target to cure HIV-1/AIDS.