Immune mechanisms of acute respiratory distress syndrome

急性呼吸窘迫综合征的免疫机制

• 批准号: 10406858
• 负责人: Brian Robert Rosborough
• 金额: $ 4.66万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10406858
• 关键词: Acidosis; Acute Lung Injury; Adult Respiratory Distress Syndrome; Alveolar; Alveolar Macrophages; Applications Grants; Biological Markers; CD14 gene; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Clinical; Clinical Trials; Critical Care; Data; Development; Disease; Edema; Endothelium; Environment; Epithelial; Epitopes; Exposure to; FCGR3B gene; Feedback; Fellowship; Flow Cytometry; Frequencies; Future; Genes; Goals; Grant; Human; Hypersensitivity; Hypoxemia; Immune; Immunology; Inflammation; Inflammatory; Inflammatory Response; Injury; Intensive Care Units; Interferon Type II; Kidney Failure; Leadership; Lung; Maintenance; Medicine; Mentors; Messenger RNA; Microarray Analysis; Molecular; Morbidity - disease rate; NK Cell Activation; Natural Killer Cells; Pathogenesis; Pathogenicity; Pathway interactions; Patient-Focused Outcomes; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Physicians; Pneumonia; Positioning Attribute; Production; Publications; Refractory; Regulation; Research; Resistance; Respiratory Failure; Risk Factors; Role; Sampling; Scientist; Sepsis; Serum; Shock; Signal Pathway; Surface; Survivors; Syndrome; System; TIS11 protein; Techniques; Technology; Testing; Training; Trauma; Universities; Writing; base; career; career development; chemokine; clinical heterogeneity; cognitive disability; cytokine; experimental study; indexing; monocyte; mortality; multidisciplinary; neutrophil; new therapeutic target; non cardiogenic pulmonary edema; novel; potential biomarker; protein expression; psychiatric disability; recruit; responsible research conduct; single-cell RNA sequencing; skills; surfactant; targeted biomarker; therapeutic target; transcriptome; transcriptome sequencing; transcriptomics; translational study

PROJECT SUMMARY/ABSTRACT The acute respiratory distress syndrome (ARDS) is a common cause of respiratory failure in the intensive care unit due to overwhelming inflammation in the lung without effective disease modifying treatments. Two sub- phenotypes of ARDS have been identified based on clinical parameters and biomarkers. Compared to hypo- inflammatory ARDS, hyper-inflammatory ARDS is characterized by shock, acidosis, and elevated inflammatory biomarkers. Patients with hyper-inflammatory ARDS have a higher mortality and may respond differentially to treatment. The overarching goal of this training grant proposal is to utilize cutting edge immunology techniques to dissect the immune mechanisms underlying ARDS development and the differentiation between the two sub-phenotypes. In our preliminary data, we identified reduced expression of tristetraprolin (TTP), a negative regulator of cytokine production, in natural killer (NK) cells of patients with ARDS. In Aim 1, we will investigate the mechanisms by which TTP regulates NK cell activation. Additionally, we will use PrimeFlow, a novel flow cytometry-based mRNA quantification technique, to determine if NK cells from patients with ARDS are refractory to negative feedback by inflammatory cytokines due to reduced TTP expression. In Aim 2, we will focus on discovery of immune mechanisms that differentiate hyper- and hypo-inflammatory ARDS. We have found increased frequency of circulating inflammatory non-classical CD14-CD16+ monocytes in hyper- inflammatory ARDS when compared to hypo-inflammatory ARDS patients. We propose using cellular indexing of transcriptomes and epitopes with sequencing (CITE-seq) to investigate signaling pathways at the single cell level in monocytes that differentiate hyper- and hypo-inflammatory ARDS. CITE-seq references single cell RNA sequencing transcriptomic data to surface expression of proteins thereby correlating the transcriptome with flow cytometric surface phenotypes. Together, aims 1 and 2 will test our overall hypotheses that NK cells in ARDS patients are resistant to negative feedback due to reduced TTP expression, and persistent activation of CD16+ non-classical monocytes determines the hyper-inflammatory phenotype of ARDS. Further delineation of immune mechanisms in the development and maintenance of ARDS will advance our understanding of its pathogenesis and identify potential therapeutic targets to modify the disease course of ARDS. The research proposed within this training fellowship will be undertaken at the University of Pittsburgh through a collaborative training environment between the Division of Pulmonary, Allergy and Critical Care Medicine and Department of Immunology to develop research skills in translational human and computational systems immunology. Formal training in the responsible conduct of research and scientific rigor are included in this training proposal, and career development through presentation of research, publication and a multi-disciplinary mentoring committee are aimed at continued progress towards developing an independent physician-scientist career.

项目概要/摘要 急性呼吸窘迫综合征（ARDS）是重症监护室呼吸衰竭的常见原因 由于肺部严重炎症而没有有效的疾病缓解治疗。两个子 ARDS 的表型已根据临床参数和生物标志物进行了鉴定。与低 炎症性 ARDS、高炎症性 ARDS 的特点是休克、酸中毒和炎症升高 生物标志物。高炎症性 ARDS 患者的死亡率较高，并且对药物的反应可能存在差异 治疗。该培训拨款提案的总体目标是利用尖端的免疫学技术 剖析 ARDS 发展背后的免疫机制以及两者之间的区别 亚表型。在我们的初步数据中，我们发现三四脯氨酸 (TTP) 的表达减少，这是一种阴性 ARDS 患者自然杀伤 (NK) 细胞中细胞因子产生的调节因子。在目标 1 中，我们将调查 TTP 调节 NK 细胞激活的机制。此外，我们将使用 PrimeFlow，一种新颖的流程 基于细胞计数的 mRNA 定量技术，以确定来自 ARDS 患者的 NK 细胞是否 由于 TTP 表达减少，难以抵抗炎症细胞因子的负反馈。在目标 2 中，我们将 专注于发现区分高炎症和低炎症 ARDS 的免疫机制。我们有 发现循环炎症非经典 CD14-CD16+ 单核细胞的频率增加 与低炎症性ARDS患者相比，炎症性ARDS患者。我们建议使用细胞索引 通过测序 (CITE-seq) 分析转录组和表位，以研究单细胞的信号传导途径 单核细胞中区分高炎症和低炎症 ARDS 的水平。 CITE-seq 参考单细胞 RNA 测序转录组数据与蛋白质表面表达，从而将转录组关联起来 具有流式细胞表面表型。目标 1 和目标 2 将共同检验我们的总体假设：NK 由于 TTP 表达减少，ARDS 患者的细胞对负反馈具有抵抗力，并且 CD16+非经典单核细胞的持续激活决定了高炎症表型 急性呼吸窘迫综合征。进一步描述 ARDS 发生和维持的免疫机制将 增进我们对其发病机制的理解并确定潜在的治疗靶点来改变疾病 ARDS 的病程。本次培训奖学金中提议的研究将在英国大学进行 匹兹堡通过肺科、过敏科和危重症科之间的协作培训环境 护理医学和免疫学系开发转化人类和免疫学的研究技能 计算系统免疫学。负责任的研究行为和科学严谨性的正式培训 包含在本培训提案中，并通过研究报告、出版物的介绍和职业发展 和一个多学科指导委员会的目的是在发展 独立的医生科学家职业。