Probing the Pathophysiology of ME/CFS through Proteomics and Metabolomics

项目总结：临床项目2：探索ME/CFS的病理生理学 深刻的疲劳，疼痛，认知困难的根本原因，以及对 ME/CFS患者的身体努力，尚不清楚。许多ME/CFS患者无法 工作是他们产生能量的最大能力，通过心肺运动测试（CPET）测量， 非常低。对于许多ME/CFS患者来说，身体或精神活动的增加会导致 这些症状被称为运动后不适。康奈尔中心调查人员先前的工作发现， 接受CPET的ME/CFS患者在一天后无法重现一项或多项客观指标，如 最大阈值强度下的氧消耗，和/或表现出血压异常，和/或 心率反应运动后不适的原因，如ME/CFS的原因，尚不清楚， 尽管它是ME/CFS患者最虚弱的症状之一。 我们的中心假设是，对运动后不适的研究提供了一个令人兴奋的机会， 获得对ME/CFS病因的新见解。我们将在几个不同的时间从病人身上采集样本 点，在运动挑战之前，当他们正在经历他们通常的症状水平时， 运动后症状加剧，使人们能够更深入地了解异常的生物功能 与疾病有关。通过结合生理数据分析代谢物、循环 炎症分子和细胞外囊泡（EV）货物在血液样品之前和之后， 运动课程，我们的目标是揭示标记物和运动后不适在ME/CFS的机制。我们 对运动可能的分子反应的广泛调查将包括炎症蛋白和免疫原性蛋白。 线粒体DNA（mtDNA）片段，血清的靶向和非靶向代谢组学，以及详细的 EV的蛋白质组学和代谢组学表征。电动汽车在运动过程中被释放到循环中， 因此可能含有生物标志物或含有在介导异常的 对ME/CFS中体力活动的反应。EV的货物包括信号蛋白、脂质、激素和RNA 这可以影响它们融合的靶细胞的生长、代谢活性和基因表达。 我们预计血清和EV的代谢组学分析具有特别高的检测潜力， ME/CFS对运动反应的特异性变化，因为代谢组在下游整合 几乎所有生理途径的影响。因此，除了常规的靶向代谢组学， 我们将介绍非靶向代谢组学分析，作为发现新的或 初级代谢或信号分子产生的意外ME/CFS相关变化， 如激素、肾上腺素或神经递质。作为这项研究的主要成果，我们希望提供 与运动后不适相关的代谢组学和蛋白质组学变化的综合数据， ME/CFS，这将使新的标志物和与这种疾病相关的机制的鉴定。

PROJECT SUMMARY: CLINICAL PROJECT 2: PROBING THE PATHOPHYSIOLOGY OF ME/CFS The underlying reason for the profound fatigue, pain, cognitive difficulties, and the peculiar response to physical effort of ME/CFS patients, is not understood. One reason that many ME/CFS patients are unable to work is that their maximal ability to produce energy, measured by cardiopulmonary exercise testing (CPET), is extremely low. For many ME/CFS patients an increase in physical or mental activity results in an increase in symptoms termed post-exertional malaise. Prior work by Cornell Center investigators found that most ME/CFS patients who undergo CPET cannot reproduce, a day later, one or more objective measures such as oxygen consumption at maximal threshold intensities, and/or exhibit abnormalities in blood pressure and/or heart rate response. The cause of post-exertional malaise, like the cause of ME/CFS, is not understood, although it is one of the most debilitating symptoms of ME/CFS sufferers. Our central hypothesis is that the study of postexertional malaise provides an exciting opportunity to obtain new insights into the etiology of ME/CFS. We will obtain samples from patients at several different time points, prior to an exercise challenge, when they are experiencing their usual levels of symptoms, and at a time of heightened symptoms, post exercise, enabling deeper insight into abnormal biological functioning associated with the disease. By analyzing, in conjunction with physiological data, metabolites, circulating inflammatory molecules, and extracellular vesicle (EV) cargo in blood samples from before and after exercise sessions, we aim to uncover markers and mechanisms of post-exertional malaise in ME/CFS. Our broad survey of possible molecular responses to exercise will include inflammatory proteins and immunogenic mitochondrial DNA (mtDNA) fragments, targeted and untargeted metabolomics of blood serum, and a detailed proteomic and metabolomic characterization of EVs. EVs are released into the circulation during exercise and could therefore contain biomarkers or contain cargo that plays an active role in mediating the abnormal response to physical activity in ME/CFS. EVs cargo includes signaling proteins, lipids, hormones, and RNAs that can influence the growth, metabolic activity, and gene expression in target cells with which they fuse. We expect that the metabolomic analyses of serum and EVs have particularly high potential to detect ME/CFS-specific changes in the response to exercise, because the metabolome integrates downstream effects from almost any physiological pathway. Therefore, in addition to conventional targeted metabolomics, we will introduce untargeted metabolomic analyses as a powerful approach toward discovery of new or unexpected ME/CFS-associated changes in primary metabolism or the production of signaling molecules such as hormones, prostaglandins, or neurotransmitters. As a primary outcome of this study, we expect to provide comprehensive data on metabolomic and proteomic changes associated with post-exertional malaise in ME/CFS that will enable identifications of novel markers and mechanisms associated with this disease.