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

项目总结/摘要 尽管有有效的抗逆转录病毒疗法（ART），艾滋病毒感染者的预期寿命缩短， 与未感染艾滋病毒的对照组相比，老年相关疾病的发病率更高。持续全身性 尽管有抑制性ART，炎症仍与严重的非AIDS发病率相关（例如，心肌 梗塞、中风、恶性肿瘤）和死亡率。最近的数据表明，白细胞介素的上游调节因子 白细胞介素-1 β（IL-1β）可能是HIV+患者心血管风险增加的主要驱动因素。 艺术压抑的人。最近的CANTOS试验已经在超过10，000例患者中证明， 用单克隆抗体canakinumab阻断体内IL-1β可显著降低心血管事件 和癌症死亡率之间的关系。我们最近进行了一项1期试验， 对HIV+ ART抑制受试者单次给药canakinumab，发现体内IL-1β阻断导致 血浆IL-1β（以及相关的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“HIV储存库”（残留病毒总量）中起关键作用 在ART抑制期间持续存在并潜在地驱动全身性炎症）。我们将使用一个公正的 一种结合多个高维数据集的综合方法，用于检验IL-1β触发 促炎反应，助长艾滋病毒免疫功能障碍和持久性。我们建议的研究将是 第一个（在HIV+或HIV未感染个体中）研究宿主遗传学与血浆IL-1β之间的联系 水平-同时测定该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例HIV+ ART抑制患者中鉴定与血浆IL-1β相关的DNA变异体。 来自艾滋病研究中心综合临床系统网络（CNICS）队列的参与者使用 定制全外显子组测序，以添加到现有的全基因组阵列数据。个人极端 然后选择表型（血浆IL-1β水平的最高和最低十分位数）进行功能验证， 目的2利用一种新的方法，同时表征RNA和蛋白质表达在单细胞 使用单细胞RNA和抗体测序（scRNA-Abseq）进行解析。最后，我们将在 体内验证与IL-1β信号传导相关的已鉴定基因，利用来自我们的1期试验的样本， 目标3中的卡那单抗。因此，我们将在功能上验证目标1的发现，并确定新的 通过研究具有独特HIV+表型的个体（极端血浆IL-1β水平和 卡那单抗治疗）。拟议的工作将确定特定的基因， 可能与IL-1β协同作用的免疫途径，确定可能 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.