Inhibition of the H3K27 demethylase UTX enhances the epigenetic silencing of HIV proviruses and induces HIV-1 DNA hypermethylation but fails to permanently block HIV reactivation.
Inhibition of the H3K27 demethylase UTX enhances the epigenetic silencing of HIV proviruses and induces HIV-1 DNA hypermethylation but fails to permanently block HIV reactivation.
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DOI：
10.1371/journal.ppat.1010014
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发表时间：
2021-10
期刊：
影响因子：
6.7
通讯作者：
Karn J
中科院分区：
文献类型：
--
作者：
Nguyen K;Dobrowolski C;Shukla M;Cho WK;Luttge B;Karn J
一种功能性治愈HIV-1的策略是“阻断和锁定”，它寻求通过表观遗传沉默来永久抑制静止HIV-1的反弹。对于HIV LTR中的二价启动子，组蛋白3赖氨酸27三甲基化（H3K27me3）和DNA甲基化都与病毒抑制相关，而H3K4三甲基化（H3K4me3）与病毒表达相关。然而，H3K27me3在通过t细胞受体激活t细胞后很容易被逆转。为了以稳定的方式抑制潜伏的HIV-1，我们敲低了主要的H3K27去甲基化酶UTX/KDM6A的表达或抑制其活性，并研究了其对T细胞中潜伏的HIV-1再激活的影响。UTX的抑制显著提高了HIV LTR的H3K27me3水平，并与DNA甲基化增加有关。GSK-J4是H3K27me3/me2去甲基化酶JMJD3/KDM6B和UTX/KDM6A的有效双重抑制剂，在患者的潜伏感染细胞中，GSK-J4有效抑制潜伏HIV-1的再激活，并通过增强DNMT3A对HIV-1的募集，诱导潜伏HIV-1的5'LTR特异性位点的DNA甲基化。尽管如此，通过表观遗传沉默抑制HIV-1需要继续使用GSK-J4治疗，并且在移除药物后迅速逆转。去除药物后，DNA甲基化也迅速消失，这表明HIV LTR的DNA去甲基化是活跃和快速的。因此，通过组蛋白和DNA甲基化诱导表观遗传沉默似乎不足以永久沉默HIV-1前病毒转录。功能性治愈HIV-1的“阻断和锁定”策略是基于一个前提，即HIV-1的永久失活可以通过对原病毒DNA的表观遗传沉默来实现。对于细胞基因，在细胞分化过程中，通过诱导特定的表观遗传修饰（包括组蛋白和DNA甲基化）实现长期沉默。在HIV-1沉默期间，观察到组蛋白甲基化和DNA甲基化，但这两组修饰都可以通过t细胞受体或有效的潜伏期逆转剂在t细胞激活后逆转。为了增强HIV-1转录的沉默，我们使用了H3K27去甲基化酶抑制剂来增加H3K27的甲基化。这反过来又导致HIV-1的DNA甲基化增强。不幸的是，尽管这种治疗有效地沉默了HIV-1并阻止了病毒的再激活，但这种沉默效果是短暂的，并且在移除药物后迅速逆转。
One strategy for a functional cure of HIV-1 is “block and lock”, which seeks to permanently suppress the rebound of quiescent HIV-1 by epigenetic silencing. For the bivalent promoter in the HIV LTR, both histone 3 lysine 27 tri-methylation (H3K27me3) and DNA methylation are associated with viral suppression, while H3K4 tri-methylation (H3K4me3) is correlated with viral expression. However, H3K27me3 is readily reversed upon activation of T-cells through the T-cell receptor. In an attempt to suppress latent HIV-1 in a stable fashion, we knocked down the expression or inhibited the activity of UTX/KDM6A, the major H3K27 demethylase, and investigated its impact on latent HIV-1 reactivation in T cells. Inhibition of UTX dramatically enhanced H3K27me3 levels at the HIV LTR and was associated with increased DNA methylation. In latently infected cells from patients, GSK-J4, which is a potent dual inhibitor of the H3K27me3/me2-demethylases JMJD3/KDM6B and UTX/KDM6A, effectively suppressed the reactivation of latent HIV-1 and also induced DNA methylation at specific sites in the 5’LTR of latent HIV-1 by the enhanced recruitment of DNMT3A to HIV-1. Nonetheless, suppression of HIV-1 through epigenetic silencing required the continued treatment with GSK-J4 and was rapidly reversed after removal of the drug. DNA methylation was also rapidly lost after removal of drug, suggesting active and rapid DNA-demethylation of the HIV LTR. Thus, induction of epigenetic silencing by histone and DNA methylation appears to be insufficient to permanently silence HIV-1 proviral transcription. The “block and lock” strategy for a functional HIV-1 cure is based on the premise that permanent inactivation of the HIV-1 can be achieved by epigenetic silencing of the proviral DNA. For cellular genes, long-term silencing is achieved during cell differentiation by the induction of specific epigenetic modifications involving histone and DNA methylation. During HIV-1 silencing, histone methylation and DNA methylation are observed, but both sets of modifications can be reversed upon activation of T-cells through the T-cell receptor or potent latency reversing agents. In an attempt to enhance silencing of HIV-1 transcription, we used an inhibitor of H3K27 demethylases to increase H3K27 methylation. This in turn led to enhanced DNA methylation of HIV-1. Unfortunately, although the treatment effectively silenced HIV-1 and prevented viral reactivation, the silencing effects were short-lived and quickly reversed after removal of the drug.
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