A chemical genetics approach for studies of HIV-1 latency

项目总结 从宿主基因组中消除整合的、具有复制能力的HIV-1前病毒尽管 抑制性抗逆转录病毒疗法(ART)是功能性治愈的主要障碍。含有这些类型的细胞 的前病毒产生边际水平的病毒产物，因此对免疫监测变得难以实现 机械装置。这种缺乏免疫系统的检测，除了它增加的生长潜力之外，由于 动态平衡增殖和克隆性扩张，延长潜伏感染细胞的寿命，产生 持久性储集层。人们对针对潜伏者的精确治疗的潜力充满了极大的热情 临床环境中的蓄水池。为了实现这一重大的生物医学目标，我们必须首先发现决定 在我们可以利用这一知识之前，存储库持久性和病毒潜伏期维护和重新激活 临床干预。虽然之前的研究已经使用了几种遗传方法来检查宿主因子 参与，他们持有内在的问题，不允许区分直接和间接的影响。 这是一个重要的问题，因为我们必须首先确定宿主因素在艾滋病毒-1潜伏期中的主要作用(S) 对照，然后阐明最合适的治疗干预方法。 在这一探索性和发展性的R21拨款申请中，我们通过以下方式绕过了前面的问题 实施一种新的化学遗传学(Dtag)方法来显著消除一组宿主的表达 染色质调节因子(组蛋白赖氨酸甲基转移酶)评估它们在HIV-1前病毒潜伏期中的作用 维护和重新激活。我们将首先在CD4+T细胞潜伏期模型中内源性地标记这些因素 重述病毒在患者中持续存在的生物学。然后，我们将选择按优先顺序进行交叉的候选人 从UT获得的潜伏期和无尿毒症患者样本在初级CD4+T细胞模型中的有效性 西南/Parkland HIV-1诊所。如果成功，我们的研究将填补我们对HIV-1潜伏期理解的空白 描述新的基础科学和阐明最合适的组蛋白赖氨酸甲基的生物学 试验性翻译研究的转移酶。在这一重点赠款申请范围之外的未来研究 将研究这些宿主染色质调节剂的临床相关性，并设计出适当的治疗方法 干预措施。

PROJECT SUMMARY Elimination of integrated, replication-competent HIV-1 proviruses from host genomes persisting despite suppressive anti-retroviral therapy (ART) is the major roadblock to a functional cure. Cells harboring these types of proviruses produce marginal levels of viral products thereby becoming refractory to immune surveillance mechanisms. This lack of detection by the immune system, in addition to its increased growth potential, due to homeostatic proliferation and clonal expansion, extend the lifespan of latently infected cells generating a persistent reservoir. There is enormous enthusiasm for the potential of precision therapies targeting the latent reservoir in clinical settings. To achieve this major biomedical goal, we must first discover host factors dictating reservoir persistence and viral latency maintenance and reactivation before we can leverage this knowledge for clinical intervention. While previous studies have used several genetic approaches to examine host factor’s involvement, they hold the intrinsic problem of not allowing to distinguish between direct and indirect effects. This is a significant issue because one must first define the host factor’s primary function(s) in HIV-1 latency control to then illuminate the most appropriate approaches for therapeutic intervention. In this exploratory and developmental R21 grant application, we circumvent previous issues by implementing a novel chemical genetics (dTAG) approach to acutely eliminate the expression of a set of host chromatin regulators (Histone Lysine Methyl Transferases) to assess their roles in HIV-1 proviral latency maintenance and reactivation. We will first endogenously tag these factors in CD4+ T cell models of latency that recapitulate the biology of viral persistence in patients. We will then select prioritized candidates for cross- validation in primary CD4+ T cell models of latency and aviremic patient samples obtained from the UT Southwestern/Parkland HIV-1 Clinic. If successful, our studies will fill a void in our understanding of HIV-1 latency biology by describing new basic science and elucidating the most appropriate Histone Lysine Methyl Transferases for pilot translational studies. Future studies beyond the scope of this focused grant application will examine the clinical relevance of these host chromatin regulators and devise appropriate therapeutic interventions.