POLYCOMB REPRESSIVE COMLEXE AS KEY REGULATORS OF HIV LATENCY AND TARGETS FOR LATENCY REVERSAL

摘要：多梳抑制复合体作为HIV潜伏期和 延迟逆转的目标 为了努力治愈艾滋病毒，消除潜伏的艾滋病毒感染的持久蓄水池的方法是 需要的。在病毒和细胞基因表达调节器的微妙平衡的强制下， 静止的逆转录病毒基因组是病毒确保其在宿主内稳定的一种非凡的方式。 操纵病毒潜伏期以允许病毒重新表达和清除病毒的可能性是领先的 慢性感染患者的HIV治愈策略。促进逆转病毒潜伏期的化学策略 通过破坏抑制性表观遗传过程，代表着朝着临床策略迈出的开创性一步 治愈艾滋病毒。幸运的是，通过表观遗传学重新激活艾滋病毒前病毒的努力已经与科学 社区对染色质调节的兴趣和对表观遗传化学物质作用的日益认识 在推动对其目标的生物学理解方面的探测。关于重新激活艾滋病毒的概念验证研究 通过抑制表观遗传调节蛋白的转录已经产生了令人振奋的结果。然而，许多人 这些化合物只表现出适度的潜伏期逆转活性，并且只有有限数量的化学工具来 实现这一目标目前是可用的。更好地理解表观遗传途径的景观 影响前驱潜伏期，以及新的化学工具，显然需要发展完全有效的潜伏期 单独或联合使用的逆转药物(LRA)，用于治愈艾滋病毒。 这项提案的总体目标是应用遗传、生化和化学生物学方法。 到潜伏期和HIV供体来源样本的细胞模型，以I)定义多梳状抑制的作用 HIV潜伏期中的复合体PRC1和PRC2，以及II)了解物质使用对Polycomb- 介导的HIV潜伏期。PrC1和PrC2是基因沉默的关键调节者，通过安装和 对抑制性H3K27me3翻译后修饰(PTM)的识别，因此，很有可能 对艾滋病毒潜伏期做出了重大贡献。我们的初步数据表明，Prc1和Prc1的成分 尚未与维持艾滋病毒潜伏期有关的PRC2，以及与Polycomb相关的 蛋白质是潜伏期的关键调节者，也是潜伏期逆转的新靶点。此外，滥用药物还有 已被证明诱导表观遗传修饰的变化，包括组蛋白甲基化，进而可能 显著影响艾滋病毒感染的吸毒者静止的CD4T细胞的表观遗传学特征。因此， 我们还旨在探讨药物滥用对静息CD4T细胞聚梳功能的影响，以及 这些环境因素对艾滋病毒潜伏期的建立和维持的影响。

ABSTRACT: POLYCOMB REPRESSIVE COMPLEXES AS KEY REGULATORS OF HIV LATENCY AND TARGETS FOR LATENCY REVERSAL In an effort to achieve a cure for HIV, approaches to eliminate the persistent reservoir of latent HIV infection are needed. Enforced by a delicate balance of regulators of viral and cellular gene expression, the persistence of the quiescent retroviral genome is a remarkable way by which the virus ensures its stability within the host. Manipulation of viral latency to allow renewed viral expression and the potential for viral clearance is a leading strategy for HIV cure in chronically infected patients. Chemical strategies that promote reversal of viral latency by disrupting repressive epigenetic processes represent a groundbreaking step towards a clinical strategy to cure HIV. Fortuitously, the effort to epigenetically reactivate HIV proviruses has intersected with the scientific community’s interest in chromatin regulation and an increased appreciation for the role of epigenetic chemical probes in driving biological understanding of their targets. Proof of concept studies in reactivating HIV transcription via inhibition of epigenetic regulatory proteins have yielded promising results. However, many of these compounds exhibit only modest latency reversing activity, and only a limited number of chemical tools to achieve this goal are currently available. A better understanding of the landscape of epigenetic pathways that influence proviral latency, as well as novel chemical tools, are clearly needed to develop fully effective latency reversing agents (LRAs) for use alone or in combination towards an HIV cure. The overarching objectives of this proposal are to apply genetic, biochemical, and chemical biology approaches to cellular models of latency and HIV+ donor-derived samples to i) define the role of the Polycomb Repressive Complexes, PRC1 and PRC2, in HIV latency and ii) understand the effect of substance use on Polycomb- mediated HIV latency. PRC1 and PRC2 are critical regulators of gene silencing through the installation and recognition of the repressive H3K27me3 post-translational modification (PTM), and hence, are well poised to make a significant contribution to HIV latency. Our preliminary data suggests that components of both PRC1 and PRC2 that have not yet been implicated in the maintenance of HIV latency, as well as Polycomb-associated proteins, are key regulators of latency and novel targets for latency reversal. Additionally, drugs of abuse have been shown to induce changes in epigenetic modifications, including histone methylation, and in turn could significantly impact the epigenetic profiles of resting CD4+ T-cells from HIV-infected drug abusers. Therefore, we also aim to explore the effect of substance abuse on Polycomb function in resting CD4+ T-cells, and the influence of such environmental factors on the establishment and maintenance of HIV latency.