Plaque and blood derived macrophages: a multi-omic assessment of CVD pathogenesis in PLWH

动脉粥样硬化性心血管疾病(ASCVD)是美国主要的死亡原因； ~艾滋病毒携带者发生心血管事件的风险增加2倍。1，2潜在的机制 PLWH中心血管疾病患病率的增加尚不完全清楚，但可能涉及 单核细胞和巨噬细胞。我们假设单核细胞将暴露在不同的炎症环境中。 在PLWH分化过程中的信号；因此，巨噬细胞将在功能和表型上 在这一人群中不同，并加剧了心血管疾病。我们的数据表明，脂体的改变可能 影响炎症、单核细胞活化和巨噬细胞分化。抗逆转录病毒治疗可能 通过减少氧化磷酸化(OXPHOS)和脂肪酸氧化(FAO)来改变血脂 ART诱导的线粒体功能障碍的后果。我们的研究发现了不同的 HIV-和HIV人群的脂类，即使这些人群中的传统脂类相似22。 与心血管疾病风险相关的生物标志物(IL-6、sCD14、TNFR1)、10-15与脂肪酸组成直接相关 PLWH中的促炎性脂类。我们还发现了促凝血剂血管的扩张 在PLWH42-44中归巢单核细胞，并将单核细胞激活与改变的脂谱19、20、45-47联系起来。 目标1：确定独特和常见的表型、转录和功能性MDM图谱 与PLWH和HIV-个体中是否存在ASCVD有关。 1A：比较我们4组MDM的表型、功能和转录图谱。 1B：将单核细胞亚群(基于CD14和CD16表达)43、49、50分类和区分为MDM 以确定子集来源是否决定了不同的功能和表型结果。 1C：从动脉粥样硬化斑块(颈动脉内膜切除术)中分离巨噬细胞，并比较 这些细胞的转录谱与我们4个参与者组的MDM中确定的谱相一致。 目的2：研究伴有和不伴有ASCVD的PLWH和HIV患者MDM激活的驱动因素。 2A：使用高级脂类组学(Lipidyzer)以及 颈动脉内膜切除术收集的动脉粥样硬化斑块的脂质组学特征。 2B：探讨体内和体外脂质谱调节对MDM基因表达和 功能能力；他汀类药物治疗将改善血脂51，从而降低MDM 激活MDM，并在体外将MDM暴露于促炎症脂类(即CERs，SAFas)将激活这些细胞。 2C：确定ART暴露对MDM谱的药物特异性影响。 目的：阐明艾滋病病毒携带者和艾滋病病毒携带者与ASCVD相关的特征。 综合、多维途径和体外途径抑制物实验探索 不同的信号驱动因子和抑制因子对MDM转录和功能谱产生级联作用。

Atherosclerotic cardiovascular disease (ASCVD) is a leading cause of death in the United States; and there is ~2-fold greater risk for CVD events for people living with HIV (PLWH).1,2 The mechanisms underlying increased CVD prevalence in PLWH are not fully understood, but likely involve heightened activation of monocytes and macrophages. We hypothesize that monocytes will be exposed to disparate inflammatory signals during differentiation in PLWH; as a result, macrophages will be functionally and phenotypically different and exacerbate CVD in this population. Our data suggest that alterations in the lipidome may influence inflammation, monocyte activation, and the differentiation of macrophages. Antiretroviral therapy may alter lipid profiles by reducing oxidative phosphorylation (OxPHOS) and fatty acid oxidation (FAO) as a consequence of ART-induced mitochondrial dysfunction. Our studies have identified differences in the lipidomes of HIV- and HIV+ populations, even when traditional lipid panels in these groups were similar22. Biomarkers associated with CVD risk (IL-6, sCD14, TNFR1),10-15 were directly related to fatty acid composition and pro-inflammatory lipid classes in PLWH. We have also identified an expansion of pro-coagulant, vascular homing monocytes in PLWH42-44 and linked monocyte activation to altered lipid profiles19,20,45-47. Aim 1: To identify unique and common phenotypic, transcriptomic, and functional MDM profiles associated with the presence or absence of ASCVD in PLWH and HIV- individuals. 1A: To compare MDM phenotypic, functional, and transcriptomic profiles across our 4 groups. 1B: To sort and differentiate monocyte subsets (based on CD14 and CD16 expression)43,49,50 into MDMs in order to determine whether subset origin determines differential functional and phenotypic outcome. 1C: To sort macrophages from atherosclerotic plaques (carotid endarterectomies) and compare the transcriptional profiles of these cells to the profiles identified in MDMs from our 4 participant groups. Aim 2: To characterize drivers of MDM activation in PLWH and HIV- persons with and without ASCVD. 2A: To characterize the plasma lipid profiles of participants using advanced lipidomics (Lipidyzer) as well as the lipidomic profiles of atherosclerotic plaques collected from carotid endarterectomies. 2B: To explore the consequences of in vivo and in vitro lipid profile modulation on MDM gene expression and functional capabilities; statin treatment will improve the lipidome51 and as a consequence, decrease MDM activation, and in vitro exposure of MDMs to proinflammatory lipids (i.e CERs, SaFAs) will activate these cells. 2C: To determine the drug-specific effects of ART exposure on MDM profiles. Aim3: To elucidate profiles associated with ASCVD in HIV- and HIV+ individuals using a comprehensive, multi-dimensional approach and in vitro pathway inhibitor experiments to explore differential drivers and inhibitors of signaling cascades on MDM transcription and functional profiles.