Reducing the HIV Reservoir: Next-Generation Latency Reversal Agents and T Cell Population Targets

项目摘要/摘要 人类免疫缺陷病毒(HIV)是导致严重发病率、死亡率和 世界各地的医疗系统。虽然抗逆转录病毒疗法(ART)提供了有效的减少病毒血症和 改善免疫功能，需要终生坚持以防止病毒反弹 宿主CD4T细胞淋巴细胞基因组中潜伏的、但具有复制能力的前病毒HIV DNA。 值得注意的是，终生治疗与经济负担、药物相互作用、耐药性和 治疗的长期效果未知。此外，与艾滋病毒疾病相关的耻辱有助于 负面的健康后果。因此，艾滋病毒的治疗具有重大的公共卫生利益。 治愈艾滋病毒的一个主要策略是使用小分子潜伏期反转剂(LRAs)来诱导病毒 在潜伏的储存库中表达，以便免疫系统能够识别和清除储存库。这个病毒式的 储存库在静息的CD4T淋巴细胞中被理解得最好。然而，艾滋病毒在其他地方的持久性本质 CD4T细胞群(“非静息”)是未知的。了解HIV在非静息的CD4T细胞中的持久性 对于确定是否需要外源性干预来耗尽CURE中的这些细胞非常重要 战略。因此，在目标1中，我们将描述静息和非静息的CD4T细胞中持续HIV的特征 来自ART抑制的HIV血清阳性参与者，使用细胞分选，一组病毒储存库测量 技术、超灵敏的抗原检测分析和近乎全长的病毒基因组测序。 为了耗尽病毒库，已经在临床研究中测试了几种LRA。然而， 到目前为止，临床上使用的LRA仅提供了少量的病毒表达诱导，并且没有一种耗尽 水库。然而，更强大的下一代LRA正在开发中，这些新型LRA的能力 在CD4T细胞中诱导足够的病毒蛋白表达以进行免疫识别和清除不是很好 已定义。此外，目前尚不清楚非静息状态的CD4T细胞，尽管转录活性更高，但 比静息的CD4T细胞，可以在没有LRA的情况下进行自发的免疫清除，或者如果这些细胞 代表了艾滋病毒持久力的另一个方面。因此，在目标2中，我们将评估小说和 有效的下一代LRA诱导HIV表达，足以在休息时清除免疫 与使用上述技术和一种新的潜伏期清除试验的非静止的CD4T细胞进行比较。 除了为艾滋病治愈研究的未来努力提供信息外，这里提出的项目完全符合 在一流研究型大学为一名多产的临床科学家制定的强化培训计划。开拓者 北卡罗来纳大学艾滋病毒治疗中心的环境和赞助商David MarGolis医学博士和Nancie Archin博士将 为培养艾滋病毒领域杰出的临床科学家提供必要的指导。

PROJECT SUMMARY/ABSTRACT The human immunodeficiency virus (HIV) is responsible for major morbidity, mortality, and cost to healthcare systems worldwide. While antiretroviral therapy (ART) provides a potent reduction of viremia and improvement in immune function, lifelong adherence is required to prevent viral rebound from the viral reservoir—latent, but replication competent proviral HIV DNA in the genomes of host CD4 T cell lymphocytes. Significantly, a lifetime of treatment is associated with financial burden, drug interactions, drug resistance, and unknown long-term effects of treatment. Furthermore, the stigma associated with HIV disease contributes to negative health outcomes. Thus, a cure for HIV is of substantial public health interest. A main strategy for HIV cure is the use of small-molecule latency reversal agents (LRAs) to induce viral expression in the latent reservoir so that the immune system can recognize and clear the reservoir. This viral reservoir is best understood within resting CD4 T lymphocytes. However, the nature of HIV persistence in other CD4 T cell populations (“non-resting”) is unknown. Understanding HIV persistence in non-resting CD4 T cells is important to determine whether exogenous interventions are necessary to deplete these cells in cure strategies. Thus, in AIM 1, we will characterize persistent HIV in resting versus non-resting CD4 T cells from ART-suppressed HIV seropositive participants using cell sorting, an array of viral reservoir measurement techniques, ultrasensitive antigen detection assays, and near-full length viral genome sequencing. In an effort to deplete the viral reservoir, several LRAs have been tested in clinical studies. However, LRAs used clinically to date only provide a small induction of viral expression, and none have depleted the reservoir. More potent next-generation LRAs are in development, however, the ability of these novel LRAs to induce sufficient viral protein expression in CD4 T cells for immune recognition and clearance is not well- defined. In addition, it is unknown whether non-resting CD4 T cells, despite being more transcriptionally active than resting CD4 T cells, can undergo spontaneous immune clearance without an LRA or if these cells represent another compartment of HIV persistence. Thus, in AIM 2 we will assess the ability of novel and potent next-generation LRAs to induce HIV expression sufficient for immune clearance in resting versus non-resting CD4 T cells using the techniques described above and a novel latency clearance assay. In addition to informing future efforts in HIV cure research, the project proposed herein fits perfectly into an intensive training plan for a productive clinician scientist at a top-tier research university. The pioneering environment of the UNC HIV Cure Center and the sponsors, David Margolis, MD and Nancie Archin, PhD will provide the mentorship necessary for the development of an outstanding clinician scientist in the HIV field.