Using Omics to Understand the Effects of Drug Abuse on HIV Latent Reservoir

项目摘要/摘要 这项研究的目的是促进对可卡因滥用(CA)与HIV潜伏的生物学联系的理解 水库(HLR)。我们假设基因调控是一个主要环节。我们提出了一种新的全基因组 RNA表达(RNAexp)研究发现从CA到HLR增加的基因调控途径，随后 通过DNA甲基化(DNaM)和中介分析来评估指定途径的联系。 随着抗逆转录病毒疗法(CART)和公共卫生战略相结合以减少艾滋病毒的成功 发达国家的艾滋病毒负担现在大多是一种慢性病，包括毒品。 用户。作为一种慢性疾病，寻找艾滋病毒的治疗方法取决于对HLR的理解和消除 (即，复制能力强但沉默的艾滋病毒前病毒整合到宿主细胞的DNA中，宿主细胞DNA重新激活 停止购物车)。可卡因是艾滋病毒携带者中滥用最频繁的非法药物之一。 CA与HIV加速进展有关，即使在CART中也是如此。可以肯定的是，我们还 观察到与坚持使用CART的CA相比，坚持使用CART的CA的HIV病毒载量(VL)下降速度较慢。使用VL 由于CA与HLR的数量有关，了解CA对HLR的生物学效应是至关重要的。 我们关注基因调控是因为它是CA和CA之间相互作用的生物学连接点 HLR。很明显，CA和CART都影响基因调控，正常基因调控被破坏 对艾滋病毒复制和传染性很重要，基因调控在 建立并重新激活HLR。据我们所知，没有研究量化可能的差异 也没有研究将CA与HLR联系起来的生物途径。因此，我们 提出以下目标： ·目标1：量化HLR并确定与CA相关的差异。 ·目的2：确定CA对HIV+个体基因调控的影响。 目的3：检测CA差异调控基因在CA和HLR之间的生物调节作用。 这项研究具有重大意义和创新性(例如，首次估计CA中的HLR，首次对 CA对CD4T细胞基因调控的影响--首次验证CA与CD4T细胞之间的基因调控 HLR)。我们按顺序设计的高风险/高回报方案非常适合R61/R33机制： 用CA确定R61期间HLR的显著差异及相关差异基因的表达 阶段，并评估基因调控作为R33阶段的中介。这项研究很可能会产生 对艾滋病毒作为CA慢性病的关键因素的重要新见解，为 针对CA的独特特征，这些特征增加了HLR，并使艾滋病毒的治疗在这一人群中更具挑战性。

PROJECT SUMMARY/ABSTRACT Goals of this study are to advance understanding of the biology linking cocaine abuse (CA) to HIV latent reservoir (HLR). We hypothesize that gene regulation is a primary link. We propose a novel genome-wide RNA expression (RNAexp) study to discover gene regulatory pathways from CA to increased HLR, followed by DNA methylation (DNAm) and mediation analyses to assess linkages for nominated pathways. With the success of combination antiretroviral therapy (cART) and public health strategies to reduce HIV incidence, much of the HIV burden in developed countries is now as a chronic disease, including among drug users. As a chronic disease, the search for an HIV cure is dependent on understanding and eliminating HLR (i.e., the replication competent but silenced HIV provirus integrated into host cells' DNA, which reactivates with cessation of cART). Cocaine is one of the most frequently abused illicit drugs among HIV+ individuals. CA is associated with accelerated HIV progression, which persists even with cART. Corroboratively, we also observe slower decline in HIV viral load (VL) among cART adherent CAs than adherent nonusers. With VL being associated with HLR quantity, understanding the biological effects of CA on HLR are critical. We focus on gene regulation because it is a biological connecting point for the interplay between CA and HLR. It is clear that both CA and cART affect gene regulation, that disruption of normal gene regulation is important for HIV viral replication and infectivity, and that gene regulation plays an important role in establishing and reactivating HLR. To our knowledge, there are no studies quantifying the likely differences in HLR by drug use overall or by CA specifically, nor studies of biological pathways linking CA to HLR. Thus, we propose the following aims: • Aim 1: Quantify the HLR and determine differences associated with CA. • Aim 2: Determine the effects of CA on gene regulation among HIV+ individuals. • Aim 3: Test CA differentially regulated genes as biological mediators between CA and HLR. This study is significant and innovative (e.g., first estimation of HLR among CAs, first genome-wide study of CA effects on gene regulation in CD4 T cells, first test of gene regulation as a mediator between CA and HLR). Our sequentially designed, high-risk / high-reward proposal, fits well with this R61/R33 mechanism: determining significant differences in HLR by CA and related differential gene expression during the R61 phase, and assessing gene regulation as mediators in the R33 phase. This study is likely to produce important new insights into key elements of HIV as a chronic disease among CAs, providing a basis for targeting unique features of CA that increase HLR and make an HIV cure more challenging in this population.