Target Host Epigenetic Regulation of HIV Proviruses to Reinforce Viral Deep Latency in Microglia

项目总结 全球艾滋病毒感染率和与艾滋病有关的死亡人数大幅下降，原因是 扩大获得联合抗逆转录病毒疗法(CART)的机会。然而，艾滋病毒流行仍然存在，而且 目前仍没有治愈艾滋病毒感染的方法。因为CART不能根除艾滋病毒，而复制能力强 病毒成为整合和沉默的前病毒，可以零星地重新激活以补充病毒 水库。作为HIV感染和持久性/潜伏期的关键解剖学“避难所”，大脑中的蓄水池 仍然是艾滋病毒治愈的主要障碍。HIV感染的单核细胞导致病毒从外周血传播 因为它们可以通过血脑屏障(BBB)迁移并分化为小胶质细胞 中枢神经系统(CNS)。小胶质细胞是持续/潜伏HIV的主要和稳定的病毒库 中枢神经系统感染，即使在CART存在的情况下也是如此。此外，感染艾滋病毒的小胶质细胞释放神经毒性 促进神经炎症并导致艾滋病毒相关神经认知障碍(手)的因素。 因此，小胶质细胞中调控HIV持续/潜伏感染的分子特征是 这对于制定有效的干预战略以控制和抑制中枢神经系统的艾滋病毒至关重要。表观遗传 监管至关重要地决定了艾滋病毒感染者的信念，并极大地促进了艾滋病毒的潜伏期。这个 该项目的重点是调查宿主表观遗传调控在促进持久性/潜伏性艾滋病毒方面的作用。 在小胶质细胞中感染，并针对其防止艾滋病毒裂解物重新激活。我们已经确认了一套朱蒙吉 含有结构域的组蛋白赖氨酸去甲基酶(KDM)作为HIV潜伏期促进基因(LPG) 优先靶向组蛋白H3K4/K36甲基化(H3K4/K36me3)，这表明了组蛋白H3K4/K36甲基化的潜在作用 组蛋白去甲基化在调节HIV潜伏期中的作用。在目标1中，我们将调查这些KDM的贡献 旨在促进人小胶质细胞和IPSC衍生的含有机物小胶质细胞(MCOs)中的HIV潜伏期。在AIM 在KDM研究的鼓舞下，我们希望继续使用我们的功能基因组和蛋白质组 通过多学科方法识别控制小胶质细胞中艾滋病毒潜伏期的其他新宿主因素的专业知识， 包括单电池CRISPR屏幕。这是一个需要解决的重要问题，以便我们能够将主机目标 在没有购物车的情况下，开发更有效的方案来加强HIV潜伏期并防止其复活 在小胶质细胞中。此外，我们还筛选了FDA批准的药物库，并确定了HIV潜伏期- 促进剂(LPA)，包括左西孟丹(LSM)。在目标3中，我们建议翻译研究以 调查FDA批准的已用于治疗和阻断HIV的临床药物的重新用途 持续/潜伏的小胶质细胞感染，可能影响表观遗传过程。

PROJECT SUMMARY The global rate of HIV infection and the number of AIDS related deaths have dramatically declined due to the expanding access to combinatorial anti-retroviral therapy (cART). However, the HIV epidemic remains and there is still no cure for HIV infection. Because cART does not eradicate HIV, while replication-competent viruses become integrated and silenced proviruses, which can be sporadically reactivated to replenish viral reservoirs. As a key anatomical “sanctuary site” for HIV infection and persistence/latency, reservoirs in brain remains a main hurdle for HIV cure. HIV-infected monocytes contribute to viral spread from the periphery blood to the brain as they can migrate across the blood brain barrier (BBB) and differentiate into microglia in the central nervous system (CNS). Microglia are a major and stable viral reservoir for persistent/latent HIV infection in the CNS, even in the presence of cART. Furthermore, HIV-infected microglia release neurotoxic factors that promote neuroinflammation and contribute to HIV-associated neurocognitive disorders (HAND). Therefore, characterization of molecular features regulating HIV persistent/latent infection in microglia is essential for developing effective intervention strategies to control and inhibit HIV in the CNS. Epigenetic regulation critically determines the faith of HIV proviruses and contributes significantly to HIV latency. The focus of this project is to investigate the role of host epigenetic regulation in promoting persistent/latent HIV infection in microglia and target it for preventing HIV lytic reactivation. We have identified a set of Jumonji domain-containing histone lysine demethylases (KDMs) as HIV latency-promoting genes (LPGs) that preferentially target histone H3K4/K36 methylation (H3K4/K36me3), which indicates the potential role of histone demethylation in regulation of HIV latency. In Aim 1, we will investigate the contribution of these KDMs to promoting HIV latency in human microglia and iPSC-derived microglia containing organoids (MCOs). In Aim 2, encouraged by the KDM studies, we would like to continue the use of our functional genomic and proteomic expertise to identify other ovel host factors governing HIV latency in microglia by multidisciplinary approaches, including single-cell CRISPR screen. This is an important issue to address so that we can novel host targets to develop more potent regimen for reinforcing HIV latency and preventing its resurrection at a cART-free setting in microglia. Furthermore, we also screened a library of FDA-approved drugs and identify HIV latency- promoting agents (LPAs), including levosimendan (LSM). In Aim 3, we propose the translational studies to investigate the repurposing of FDA-approved drugs already used in clinic for treating and blocking HIV persistent/latent infection in microglia, which likely affect epigenetic processes.