Host genetic predictors of plasma IL-1b levels and pharmacogenomics of in vivo IL-1b blockade during treated HIV disease

项目摘要/摘要 尽管有效的抗逆转录病毒疗法(ART)，艾滋病毒感染者的预期寿命和 与未感染艾滋病毒的对照组相比，与老龄化相关的疾病的发生率更高。持久性全身性 尽管ART受到抑制，但炎症与严重的非艾滋病发病率(例如心肌梗死)有关 脑梗塞、中风、恶性肿瘤)和死亡率。最近的数据表明，白细胞介素2的上游调节因子 (IL)-6，即白细胞介素1β(IL-1β)，可能是艾滋病毒感染者心血管风险增加的主要驱动因素 被艺术压抑的个体。最近的Cantos试验现在已经在10,000多名个体中证明了 用单抗Canakinumab阻断体内IL-1β可显著减少心血管事件 以及普通人群中的癌症死亡率。我们最近进行了一项1期试验，对 单剂Canakinumab对艾滋病毒+抗逆转录病毒治疗受试者的抑制作用，并发现在体内IL-1β阻断导致 血浆IL-1、β及相关全身炎症标志物血浆IL-6和 高敏C反应蛋白)、血管炎症和单核细胞活化。此外，我们的体外 数据表明，IL-β在维持“艾滋病毒蓄水池”(残留病毒总量)方面起着关键作用 这在ART抑制期间持续存在，并可能导致全身炎症)。我们将使用不偏不倚的 结合多个高维数据集的集成方法来测试IL-1β触发的假设 促进艾滋病毒免疫功能障碍和持久性的促炎反应。我们建议的研究将是 第一个探索宿主遗传学和血浆IL-1β之间联系的人(艾滋病毒阳性或未感染艾滋病毒者) 水平-同时检测这一途径中的几种促炎细胞因子，包括IL-6和 IL-18。在目标1中，我们将在1,000名HIV+ART抑制的患者中鉴定与血浆IL-1β相关的dna变体 来自艾滋病研究中心综合临床系统(CNICs)队列的参与者使用 定制整个外显子组测序，以添加到现有的全基因组阵列数据。极端的个人 然后将选择表型(血浆IL-1β水平的最高和最低十分之一)进行功能验证 目的2使用一种新的方法同时表征单细胞的RNA和蛋白质表达 使用单细胞RNA和抗体序列的分辨率(scRNA-Abseq)。最后，我们将在中执行函数 利用我们的第一阶段试验的样本，对已识别的与IL-1β信号相关的基因进行活体验证 目标3中的Canakinumab。因此，我们将从功能上验证目标1的发现，并确定新的 通过研究具有独特艾滋病毒+表型(极端血浆IL-1β水平及以后)的个体的基因/途径 分别在AIMS 2和3中进行Canakinumab治疗)。拟议的工作将识别特定的基因和 可能与IL-1β协同作用的免疫途径，识别可能是 广泛适用于治疗HIV+和非HIV人群中的炎症相关疾病。

PROJECT SUMMARY/ ABSTRACT Despite effective antiretroviral therapy (ART), HIV-infected individuals have reduced life expectancy and a higher incidence of aging-associated diseases compared to HIV-uninfected controls. Persistent systemic inflammation despite suppressive ART has been associated with serious non-AIDS morbidity (e.g., myocardial infarction, stroke, malignancy) and mortality. Recent data suggests that an upstream regulator of interleukin (IL)-6, interleukin-1 beta (IL-1β), may be the major driver of increased cardiovascular risk observed in HIV+ ART-suppressed individuals. The recent CANTOS trial has now demonstrated in over 10,000 individuals that in vivo IL-1β blockade with the monoclonal antibody canakinumab significantly reduced cardiovascular events and cancer mortality in the general population. We have recently performed a phase 1 trial administering a single dose of canakinumab to HIV+ ART-suppressed participants and found that in vivo IL-1β blockade led to significant reductions in plasma IL-1β (as well as associated systemic inflammatory markers plasma IL-6 and high sensitivity C-reactive protein), vascular inflammation, and monocyte activation. Furthermore, our ex vivo data suggests that IL- β plays a critical role in maintaining the “HIV reservoir” (the total amount of residual virus that persists during ART suppression and potentially drives systemic inflammation). We will use an unbiased integrated approach that combines several high dimensional datasets to test the hypothesis that IL-1β triggers the proinflammatory response that fuels HIV immune dysfunction and persistence. Our proposed study will be the first (in HIV+ or HIV-uninfected individuals) to pursue the link between host genetics and plasma IL-1β levels – while simultaneously assaying several proinflammatory cytokines in this pathway, including IL-6 and IL-18. In Aim 1 we will identify DNA variants associated with plasma IL-1β in 1,000 HIV+ ART-suppressed participants from the Center for AIDS Research Network of Integrated Clinical Systems (CNICS) cohort using custom whole exome sequencing to add to existing genomewide array data. Individuals with extreme phenotypes (highest and lowest deciles of plasma IL-1β levels) will then be selected for functional validation in Aim 2 using a novel approach that simultaneously characterizes RNA and protein expression at single cell resolution using single cell RNA and antibody sequency (scRNA-Abseq). Finally, we will perform functional in vivo validation of identified genes associated with IL-1β signaling, leveraging samples from our phase 1 trial of canakinumab in Aim 3. Therefore, we will functionally validate findings from Aim 1 as well as identify novel genes/pathways by studying individuals with unique HIV+ phenotypes (extreme plasma IL-1β levels and after canakinumab treatment, respectively) in Aims 2 and 3. The proposed work will identify specific genes and immune pathways that may act synergistically with IL-1β, identifying novel therapeutic targets that may be broadly applicable for the treatment of inflammation-associated diseases in HIV+ and non-HIV populations.