Spingolipids in HIV-associated Chronic Obstructive Pulmonary Disease

项目总结： 艾滋病毒携带者(PLWH)生存率的提高导致了几种慢性病的更高患病率 包括慢性阻塞性肺疾病(COPD)，估计影响3%-23%。在以下方面存在重大差距 知识是无法识别风险人群，以及对艾滋病毒为什么会增加COPD风险的有限理解 与吸烟状态无关。在我们最近发表的报告中，我们确定了172种独特的血浆分析器 将HIV+COPD患者与其HIV+匹配的对照组进行比较。我们确定了其中的17个 经LC-MS/MS鉴定为脂类的代谢物，包括甘油二酯和几种鞘磷脂。神经鞘脂类是 细胞信号转导的重要参与者，作为平衡细胞增殖、支持和反对的变阻器发挥作用 细胞凋亡状态和炎症过程。在HIV阴性的COPD患者中，神经鞘脂脂， 尤其是神经酰胺，与COPD的发病机制有关，并被认为是疾病的生物标志物。我们 建议在我们以前工作的基础上，利用我们的样本库来定义分子 PLWH在发展为COPD的过程中的签名，并揭示疾病的潜在机制。我们 假设一种独特的脂质生物特征与慢性阻塞性肺疾病易感性有关 与发病机制直接相关。 在这个拟议的研究项目中，我们将使用最先进的代谢组和蛋白质组方法 目的：探讨人类免疫缺陷病毒(HIV)患者血脂谱与肺功能加速下降的关系。我们的 研究利用了我们团队的专业知识(COPD生物标记物的发现；代谢组学和 计算分析；HIV感染中的COPD)和匹兹堡肺病提供的独特机会 艾滋病毒，温哥华肺艾滋病毒和抗逆转录病毒治疗的战略时机(START)队列。我们的第一个方法 目的是使用支气管肺泡灌洗(BAL)样本确定与HIV-COPD相关的肺特异性脂类分布 来自表现良好的匹兹堡和温哥华队列。我们从我们出版的作品中预计会有很多 与HIV-COPD相关的血脂将是肺特有的。揭开关键的规范路径和 与HIV-COPD相关的生物学过程我们将使用假设驱动的基于蛋白质组学的方法 包括数据独立采集质谱仪(DIA-MS)和顺序加窗采集 BAL的所有理论碎片离子质谱图(SWATH-MS)，数据分析包括基因 本体论。鞘磷脂的下游信号通路，如细胞增殖、促凋亡和抗凋亡 国家和炎症进程，将作为我们的假设驱动的重点。然后我们将验证肺 血浆中存在的预测使用血浆加速肺功能下降的特定脂谱 在我们的纵向肺活量测定子研究中，来自1,026名开始受试者的样本。

Project Summary: Improved survival in persons living with HIV (PLWH) has led to a higher prevalence of several chronic illnesses including chronic obstructive pulmonary disease (COPD), which affects an estimated 3-23%. A major gap in knowledge is the inability to identify those at risk and a limited understanding of why HIV increases COPD risk independent of smoking status. In our recent published report, we identified 172 unique plasma analytes that differentiated HIV+ individuals with COPD compared to their HIV+ matched controls. We identified 17 of these metabolites to be lipids by LC-MS/MS, including diacylglycerol and several sphingolipids. Sphingolipids are important participants in cell signaling and function as a rheostat balancing cell proliferation, pro- and anti- apoptotic states and the inflammatory process. In HIV negative individuals with COPD, sphingolipids, particularly ceramide, are linked to COPD pathogenesis and serve as putative biomarkers of disease. We propose to build upon our previous work and leverage our sample repositories to define the molecular signature of PLWH en route to developing COPD and uncover underlying mechanisms of disease. We hypothesize that a unique lipid biosignature is associated with HIV-COPD susceptibility in PLWH and is directly linked to pathogenesis. In this proposed research project, we will use state-of-the-art metabolomic and proteomic approaches to determine the relationship between lipid profiles with accelerated lung function decline in HIV. Our research takes advantage of our team's expertise (biomarker discovery in COPD; metabolomics and computational analysis; COPD in HIV infection) and the unique opportunity presented by the Pittsburgh Lung HIV, Vancouver Lung HIV and Strategic Timing of Antiretroviral Treatment (START) cohorts. Our first approach is to identify lung specific lipid profiles associated with HIV-COPD using bronchoalveolar lavage (BAL) samples from the well-phenotyped Pittsburg and Vancouver cohorts. We anticipate from our published work that many of the plasma lipids associated with HIV-COPD will be lung specific. To unravel key canonical pathways and biological processes linked to HIV-COPD we will employ a hypothesis-driven proteomic-based approach comprising data-independent acquisition mass spectrometry (DIA-MS) with Sequential Windowed Acquisition of All Theoretical Fragment Ion Mass Spectra (SWATH-MS) of BAL, with data analysis including gene ontology. Downstream signaling pathways of sphingolipids, such as cell proliferation, pro- and anti-apoptotic states and the inflammatory process, will serve as our hypothesis-driven focus. We will then validate lung specific lipid profiles that are present in plasma and predict accelerated lung function decline using plasma samples from the 1,026 START subjects in our longitudinal spirometry substudy.