Multiomic, mass spectrometry-based analysis of dried blood for deep phenotyping of sepsis

败血症，其特征是由宿主不受控制的反应导致危及生命的器官功能障碍 感染是医院的主要死亡原因，每年在美国造成25万人死亡。 花费了超过200亿美元的病人护理费用。败血症的死亡率在两年内接近50%，这已经 对严重新冠肺炎(一种病毒性脓毒症)的康复患者具有深远的意义。因为败血症是一种 多器官疾病，循环蛋白和代谢物的定量是确定脓毒症的关键 以及它的长期影响，一些这样的检测(乳酸、降钙素原)已经成为护理的标准。 到目前为止，大多数研究，特别是大量关于新冠肺炎脓毒症的研究，都描述了血浆和 与全血相比，血清充满了变异性，掩盖了红细胞的重要作用 和其他类型的细胞。为此，杜克大学的生物库是来自COVID的库样本- 重症监护病房(ICU)和冠状病毒感染后门诊的19名脓毒症患者一直在使用 容量吸收微量采样(VAMS)在Neoteryx mitra尖端采集和存储全血标本的应用 以供将来的经济分析之用。这个应用程序的首要目标是开发和验证多种方法 对Mitra TIPS中的蛋白质和代谢物进行基于质谱学的定量，全面地 败血症病理生物学简介。在R21阶段，我们将开发和验证基于质谱学的方法 对于蛋白质的非靶向定量和翻译后修饰(磷酸化， 全血中的糖基化)，并用于多种代谢物类别的靶向定量。我们将发展 这些方法的实验室间翻译的参考材料和标准操作程序。在……里面 在R33期，我们将分析来自600多个患者时间点的蛋白质、PTM和代谢物 危重脓毒症和新冠肺炎后队列中包含纵向样本的生物储存库，我们将 将这些数据与精心策划的临床数据集整合在一起。这些目标的实现将建立血液的效用 VAMS采样用于未来的脓毒症研究，并将创建高度精选的临床和深度多组学数据集 为未来的假说生成做准备。

Sepsis, which is characterized by life threatening organ dysfunction caused by an uncontrolled host response to infection, is the leading cause of death in hospitals and is responsible for >250,000 deaths per year in the U.S. at the cost of over $20 billion in patient care. Sepsis has a mortality rate nearing 50% at 2 years, which has profound implications for patients recovering from severe COVID-19, a form of viral sepsis. Because sepsis is a multi-organ disease, the quantification of circulating proteins and metabolites is central to the profiling of sepsis and of its long-term effects, and some such assays (lactate, procalcitonin) have become the standard of care. To date, the majority of studies, and particularly the large number in COVID-19 sepsis, have profiled plasma and serum, which, compared to whole blood is fraught with variability and belies the important roles of the erythrocyte and other cell-types. To this end, biorepositories at Duke University, which are banking samples from COVID- 19 sepsis patients in the intensive care unit (ICU) and in a post-COVID outpatient clinic, have been utilizing volumetric absorptive microsampling (VAMS) on Neoteryx Mitra tips to collect and store whole blood specimens for future ‘omic analyses. The overarching goals of this application are to develop and validate multiomic methods for mass spectrometry-based quantification of proteins and metabolites from Mitra tips that comprehensively profile sepsis pathobiology. In the R21 phase, we will develop and validate mass spectrometry-based methods for the non-targeted quantification of proteins and post-translational modifications (phosphorylation, glycosylation) in whole blood, and for targeted quantification of numerous metabolite classes. We will develop reference materials and standard operating procedures for inter-laboratory translation of these approaches. In the R33 phase, we will analyze proteins, PTMs and metabolites from over 600 patient timepoints from biorepositories containing longitudinal samples in critically ill sepsis and post-COVID-19 cohorts, and we will integrate these data with well-curated clinical datasets. Completion of these aims will establish the utility of blood sampling by VAMS for future sepsis studies and will create a highly curated clinical and deep multiomic dataset for future hypothesis generation.