Monitoring pro-resolving leukocyte responses in peripheral blood predicts clinical severity during sepsis

监测外周血中促溶解白细胞反应可预测脓毒症期间的临床严重程度

• 批准号: 10541851
• 负责人: Bruce D Levy
• 金额: $ 22.16万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541851
• 关键词: Address; Affect; Agonist; Anti-Inflammatory Agents; Biological Assay; Biological Models; Blood; Blood Volume; Blood capillaries; Blood specimen; Cause of Death; Cells; Cessation of life; Clinical; Clinical Data; Clinical Management; Clinical assessments; Collection; Communities; Critical Care; Critical Illness; Data; Data Set; Defect; Devices; Diagnostic; Disease; Disease Progression; Elements; Experimental Models; Family; Fingers; Flow Cytometry; Functional disorder; Future; Generations; Goals; Health; Homeostasis; Hospitals; Human; Immune; Immune response; Infection; Inflammation; Inflammatory; Inflammatory Response; Integration Host Factors; Intervention Studies; Length of Stay; Leukocytes; Link; Maps; Mass Spectrum Analysis; Measures; Mediating; Mediator; Methodology; Microfluidics; Modeling; Molecular; Monitor; Mus; National Institute of General Medical Sciences; Organ; Organ failure; Pathway interactions; Patient Selection; Patient-Focused Outcomes; Patients; Peripheral; Phagolysosome; Phase; Phenotype; Phosphorylation; Population; Process; Protein Analysis; Proteomics; Recovery; Registries; Reproducibility; Research; Resolution; Risk; Sampling; Sepsis; Severities; Shock; Signal Pathway; Signal Transduction; Stratification; System; Technology; Testing; Time; Tissues; Venous; White Blood Cell Count procedure; Whole Blood; Work; biobank; cell type; cellular transduction; clinical biomarkers; clinical predictors; clinical prognostic; counterregulation; design; immune function; improved; innovation; leukocyte activation; longitudinal analysis; mortality; neutrophil; new therapeutic target; novel; novel therapeutic intervention; organ injury; patient health information; patient subsets; peripheral blood; phosphoproteomics; prognostic; prognostic indicator; protein expression; receptor; response; restoration; scale up; septic patients

Abstract Sepsis is characterized by immune cell dysfunction, which is linked to the pathophysiology of the disease with consequent organ injury and increased associated mortality. In experimental sepsis in mice, regulating leukocyte function decreases organ injury and lessens the severity of the infection. Assessment of peripheral blood leukocyte activation and function may serve as prognostic clinical biomarkers and inform new therapeutic strategies for the management of human sepsis and inflammatory diseases. Resolution of inflammation is an active process with anti-inflammatory and pro-resolving mechanisms. There are several families of resolution mediators, termed “specialized pro-resolving mediators (SPMs)”, that serve as agonists at cognate receptors to transduce cell-type specific responses on leukocytes and in experimental model systems these SPMs decrease the severity and duration of sepsis. Recently, in microliter quantities of peripheral blood, we determined that functional characterization of leukocyte responses was more informative than leukocyte counting for clinical assessment of the trajectory of critical illness. In response to the NIGMS NOT-GM-19-054 “Exploring the Scientific Value of Existing or New Sepsis Human Biospecimen Collections”, the proposed proof of concept and scale-up studies are designed to determine the scientific value of our existing and ongoing Registry of Critical Illness biorepository of human sepsis biospecimens with associated patient health record data. These biospecimens are collected at presentation, hospital day 3 and hospital 7 to align with the clinical trajectory of the patient, linked to clinical datasets and represent transformative potential for sepsis endotyping/stratification. Here, we propose the use of isodielectric separation as a contemporary cutting-edge technology in the analysis of these biospecimens to uncover leukocyte activation during the upslope and resolution of patient’s immune responses that will test the hypothesis: Sepsis hyper- inflammatory responses result from defective endogenous resolution mechanisms and that these defects are evident in peripheral blood leukocyte activation and functional responses to SPMs. To address this hypothesis, we plan four specific aims. In the R21 phase, (1) demonstrate reproducible isolation and functional analysis of neutrophils from capillary blood, and (2) demonstrate ability to obtain deep proteomic and phosphoproteomic profiles from small numbers of neutrophils. In the R33 phase, (3) longitudinal analysis of SPM pathway activation during sepsis resolution, and (4) inference and model generation and testing with ex vivo assays. The goals of this application are to develop an improved understanding of the resolution defects that unleash unrestrained inflammation of sepsis with a longer term goal for new therapeutic targets and prognostic indicators of disease progression. Important products of this application include provision of guidance on the best practices for collecting, utilizing, and analyzing human biospecimens to maximize their value for the entire sepsis research community.

抽象的 脓毒症的特点是免疫细胞功能障碍，这与该疾病的病理生理学有关 随之而来的器官损伤和相关死亡率的增加。在小鼠实验性败血症中，调节 白细胞功能可减少器官损伤并减轻感染的严重程度。周边评估 血液白细胞活化和功能可作为预后临床生物标志物并为新的疾病提供信息 管理人类脓毒症和炎症性疾病的治疗策略。分辨率 炎症是一个具有抗炎和促消退机制的活跃过程。有几个 解决调解员家族，称为“专业促解决调解员（SPM）”，充当激动剂 在同源受体上转导白细胞和实验模型中的细胞类型特异性反应 这些 SPM 系统可降低脓毒症的严重程度和持续时间。最近，以微升计 外周血，我们确定白细胞反应的功能特征更具信息性 比白细胞计数更适合临床评估危重疾病的发展轨迹。响应 NIGMS NOT-GM-19-054“探索现有或新的脓毒症人类生物样本收藏的科学价值”， 拟议的概念验证和扩大研究旨在确定我们的科学价值 现有和正在进行的人类败血症生物样本的危重疾病生物储存库登记处以及相关 患者健康记录数据。这些生物样本是在就诊时、住院第 3 天和住院第 7 天收集的 与患者的临床轨迹保持一致，与临床数据集相关联并代表变革潜力 用于脓毒症内分型/分层。在这里，我们建议使用等介电分离作为现代技术 分析这些生物样本以揭示白细胞激活过程中的尖端技术 患者免疫反应的上升和消退将检验这一假设：脓毒症过度 炎症反应是由有缺陷的内源性消解机制引起的，并且这些 外周血白细胞激活和对 SPM 的功能反应存在明显缺陷。 为了解决这个假设，我们计划了四个具体目标。在 R21 阶段，(1) 证明可重现 从毛细血管血中分离和功能分析中性粒细胞，以及（2）证明获得深部 来自少量中性粒细胞的蛋白质组和磷酸化蛋白质组谱。在R33相中，(3)纵向 败血症缓解期间 SPM 通路激活的分析，以及 (4) 推理和模型生成以及 通过离体测定进行测试。该应用程序的目标是加深对 解决导致脓毒症不受控制的炎症的缺陷，并以新疗法为长期目标 疾病进展的目标和预后指标。该应用的重要产品包括 提供有关收集、利用和分析人类生物样本的最佳实践的指导 最大限度地提高其对整个脓毒症研究界的价值。