Pediatric Sepsis Biorepository and Clinical Database

项目摘要 脓毒症被广泛定义为由感染引起的危及生命的器官功能障碍。然而， 脓毒症在个体之间和个体内变化很大。这种个体间和个体内的异质性 反映了宿主-病原体相互作用引起的复杂的、不断发展的病理生物学， 患者特征和正在进行的医学治疗。这是极不可能的，一个单一的，甚至一个小的 一组生物标志物将以足够的准确度来描述个体的病理生物学特征，以解锁精确度 败血症的治疗方法因此，面临的挑战是确定和准确描述相关的 败血症的表型，并制定富集策略，设计和测试新的靶向治疗。为此 项目，我们建议测试新方法的可行性，收集，处理和分析生物数据 代表脓毒症病程早期对感染的免疫炎症代谢反应，并将其与 在一些实施例中，可以将信息与来自电子健康记录（EHR）的基于相关器官功能障碍的表型数据相关联。 我们组建了一个多学科团队，由转化免疫学、儿科学和免疫学领域的专家组成。 急诊医学、重症监护和数据科学。我们将测试收集和处理的可行性 在复苏前阶段，不同体积的血液样本用于深度表型分析，这是一个关键的 时间点，以及48小时后，以捕捉复苏后的动态变化。我们会用这些样本 使用高维流式细胞术表征功能性免疫炎症代谢生物学， 跟踪淋巴细胞增殖、凋亡、耗竭和细胞因子的表面和细胞内标记物 生产此外，我们将扩展流式细胞术技术，以进行生物能量评价，并将 使用O-link对血浆免疫炎症代谢组学反应进行蛋白质组学评价。 该项目将利用三个研究中心广泛的临床研究基础设施。在 此外，研究人员将获得创新的生物测定在CHOP，这将使我们的 协作团队从重症和复杂患者中收集关键生物表型。在R21阶段， 我们将完成一项单中心试点研究，以测试收集、处理和分析 在脓毒症早期阶段进行深度表型分析的最佳血液标本，将其与EHR注册表联系起来 数据，并开发和测试数据管道，以表征患者的基于器官功能障碍的临床 表型在R33阶段，我们将把样本采集扩展到三个研究中心，并进行 概念分析，以确定是否存在免疫炎症代谢组学模式的差异， 发展为任何一种基于高危器官功能障碍的脓毒症表型的儿童。识别相关 R33阶段的生物模式可以导致未来的假设驱动的机制研究， 发展靶向治疗。

PROJECT SUMMARY Sepsis is broadly defined as life-threatening organ dysfunction caused by infection. However, the experience of sepsis varies widely between individuals and within individuals. Such inter- and intra-individual heterogeneity reflects complex, evolving pathobiology induced by host-pathogen interactions and modified by pre-existing patient characteristics and ongoing medical treatments. It is highly unlikely that a single—or even a small panel—of biomarkers will characterize an individual’s pathobiology with enough accuracy to unlock precision medicine approaches in sepsis. The challenge, therefore, is to identify and accurately characterize relevant phenotypes of sepsis and develop enrichment strategies to design and test novel targeted therapies. For this project, we propose testing the feasibility of novel approaches to collect, process, and analyze biologic data representing the immuno-inflammatory-metabolic response to infection early in the sepsis course and link this information to relevant organ dysfunction-based phenotype data from the electronic health record (EHR). We have assembled a multi-disciplinary team composed of experts in translational immunology, pediatric emergency medicine, critical care, and data science. We will test the feasibility of collecting and processing blood samples of different volumes for deep phenotyping at the pre-resuscitation phase, which is a critical timepoint, as well as 48 hours later to capture dynamic changes post-resuscitation. We will use these samples to characterize the functional immuno-inflammatory-metabolic biology using high dimensional flow cytometry to track surface and intracellular markers of lymphocyte proliferation, apoptosis, exhaustion, and cytokine production. In addition, we will extend flow cytometric techniques to perform bioenergetic evaluation, and will perform proteomic evaluation of the plasma immuno-inflammatory-metabolomic response using O-link. This project will take advantage of extensive clinical research infrastructure at the three study sites. In addition, the investigators will have access to innovative biologic assays at CHOP which will allow our collaborative team to glean key biologic phenotypes from critically ill and complex patients. In the R21 phase, we will complete a single-center pilot study to test the feasibility of collecting, processing, and analyzing the optimal blood specimens for deep phenotyping in the early phase of sepsis, linking them to the EHR registry data, and developing and testing a data pipeline to characterize the patients’ organ dysfunction-based clinical phenotypes. In the R33 phase, we will expand the sample collection to three sites and perform a proof-of- concept analysis to determine if there are differences in the immuno-inflammatory-metabolomic patterns of the children who develop either of the high-risk organ dysfunction-based sepsis phenotypes. Identifying relevant biological patterns in the R33 phase can then lead to future hypothesis-driven mechanistic studies and the development of targeted therapies.