Probing the Pathophysiology of ME/CFS through Proteomics and Metabolomics

项目摘要：临床项目2：探讨ME/CFS的病理生理学 导致严重疲劳、疼痛、认知困难和对 ME/CFS患者的体力活动尚不清楚。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)来衡量，是 非常低。对于许多ME/CFS患者来说，体力或精神活动的增加会导致 这些症状被称为劳后不适。康奈尔中心调查人员之前的工作发现，大多数 接受CPET的ME/CFS患者不能在一天后复制一种或多种客观措施，如 最大阈值强度的氧气消耗，和/或表现出血压和/或异常 心率反应。运动后不适的原因，就像ME/CFS的原因一样，尚不清楚， 虽然这是ME/CFS患者最虚弱的症状之一。 我们的中心假设是，对劳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患者身上获取样本 积分，在锻炼挑战之前，当他们经历他们通常水平的症状时，一次 运动后症状加重，使人们能够更深入地了解生物功能异常 与这种疾病有关。通过分析，结合生理数据，代谢产物，循环 前后血样中炎性分子和细胞外小泡(EV)的含量 通过运动训练，我们的目标是揭示ME/CFS运动后不适的标志和机制。我们的 对运动可能的分子反应的广泛调查将包括炎性蛋白和免疫原性 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 EVS的蛋白质组学和代谢组学特征。电动汽车在运动和运动中被释放到循环中 因此可能含有生物标志物或含有在调节异常的过程中发挥积极作用的货物 在ME/CFS中对体力活动的反应。EVS货物包括信号蛋白、脂质、激素和RNA 这会影响它们与之融合的目标细胞的生长、代谢活动和基因表达。 我们预计血清和EVS的代谢分析具有特别高的检测潜力 ME/CFS在运动反应中的特异性变化，因为代谢组整合在下游 几乎所有生理途径的影响。因此，除了传统的靶向代谢组学之外， 我们将引入非靶向代谢分析，作为发现新的或新的 初级代谢中与ME/CFS相关的意外变化或信号分子的产生 如激素、前列腺素或神经递质。作为这项研究的主要结果，我们预计将提供 与运动后不适相关的代谢和蛋白质组学变化的综合数据 ME/CFS，这将使识别与这种疾病相关的新标记和机制成为可能。

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.