Project 3: Targeting linear ubiquitination to attenuate inflammation and promote repair after viral pneumonia

项目3：靶向线性泛素化以减轻病毒性肺炎后的炎症并促进修复

基本信息

批准号：
10269676
负责人：
GR Scott Budinger
金额：
$ 54.4万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-15 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10269676
关键词：
2019-nCoV Acute Adult Respiratory Distress Syndrome Alveolar Alveolar Macrophages Antiviral Therapy Attenuated Cell physiology Cells Cessation of life Clinical Complex Data Development Enrollment Epithelial Epithelial Cells Failure Functional disorder Funding Opportunities Genes Genetic Transcription Heme Hypoxemia Infection Inflammation Inflammatory Inflammatory Response Influenza A virus Infrastructure Injury Instruction Interferon Type I Iron Regulatory Protein 2 Ligase Liquid substance Lung Measures Mediating Microbiology Molecular NF-kappa B Organ Outcome Paper Patients Phenotype Physiological Play Pneumonia Proteins Recovery Regulation Reporting Research Resolution Respiratory Failure Role Signal Transduction Supportive care Techniques Therapy trial Ubiquitin Ubiquitination Viral Pneumonia Virus Diseases alveolar epithelium community acquired pneumonia cytokine epithelial repair experimental study improved outcome individualized medicine injury and repair insight iron oxide lung injury lung repair macrophage monocyte mortality mouse model pathogen pneumonia treatment prevent repair function repaired response transcription factor ubiquitin ligase ubiquitin-protein ligase

项目摘要

In this PPG, all of the Projects and Cores consider the persistence of respiratory failure and the development of multiple organ dysfunction in patients with ARDS as a failure of normal mechanisms of inflammation resolution and lung repair. This hypothesis is clinically supported by a recent analysis of patients enrolled in the ARDSnet where a “hyerinflammatory” endotype of ARDS was associated with poor clinical outcomes including death. The expression of many genes encoding inflammatory cytokines is regulated by the transcription factor NF-B. Hence, understanding how NF-B activity is controlled over the course of lung injury and repair is likely to provide mechanistic insights into the pathobiology of successful or failed lung repair after injury. The Linear Ubiquitin Assembly Complex (LUBAC) is an E3 ubiquitin ligase that performs Met-1 ubiquitination necessary for NF-B activation. In preliminary experiments, we observed that epithelial specific deletion of a modulatory component of LUBAC (HOIL-1L) prevents NF-B signaling and improves outcomes in a murine model of influenza A virus infection. We present additional preliminary data that support our hypothesis that LUBAC promotes persistent NF-B signaling in the alveolar epithelium and monocyte-derived alveolar macrophages to inhibit lung repair after influenza A induced lung injury. Specific Aim 1. To determine whether inhibiting LUBAC-mediated NF-B activation in the lung epithelium during recovery from influenza A infection accelerates lung repair. We will perform timed experiments in which we will genetically inhibit LUBAC mediated NF-B signaling during recovery from influenza A infection and use complementary physiologic and molecular techniques to measure lung repair. Specific Aim 2. To determine whether activation of LUBAC mediated NF-B activation in the lung epithelium directs reparative phenotypes in monocyte-derived macrophages. We will determine whether the inflammatory phenotype of these cells is driven by the changing microenvironment—specifically the lung epithelium, or whether these changes are cell autonomous within the macrophage. Specific Aim 3. To determine whether increased expression of NF-B-dependent inflammatory genes in alveolar macrophages and BAL fluid from patients with severe pneumonia is associated with 30 day mortality. We will take advantage of the infrastructure provided by the Successful Clinical Response in Pneumonia Therapy (SCRIPT) trial to determine whether there is an association between the expression of NF- B target genes in alveolar macrophages and BAL fluid cytokines with clinical outcomes.

在此 PPG 中，所有项目和核心都考虑了呼吸衰竭的持续性以及呼吸衰竭的发展 ARDS 患者的多器官功能障碍是正常炎症消退机制的失败最近对 ARDSnet 中登记的患者进行的分析在临床上支持了这一假设。 ARDS 的“过度炎症”内型与包括死亡在内的不良临床结果相关。许多编码炎症细胞因子的基因的表达受转录因子 NF-κB 的调节。因此，了解 NF-κB 活性在肺损伤和修复过程中如何受到控制可能会提供帮助。对损伤后肺修复成功或失败的病理学机制的见解。线性泛素组装复合物 (LUBAC) 是一种 E3 泛素连接酶，可进行 Met-1 泛素化在初步实验中，我们观察到上皮特异性缺失a。 LUBAC (HOIL-1L) 的调节成分可防止 NF-κB 信号传导并改善小鼠的结果我们提供了额外的初步数据来支持我们的假设： LUBAC 促进肺泡上皮和单核细胞衍生肺泡中持续的 NF-κB 信号传导甲型流感引起的肺损伤后巨噬细胞抑制肺修复。具体目标 1. 确定是否抑制肺上皮中 LUBAC 介导的 NF-κB 激活在从甲型流感感染恢复期间，我们将进行定时实验。我们将在甲型流感感染恢复期间从基因上抑制 LUBAC 介导的 NF-κB 信号传导，并且使用互补的生理学和分子技术来测量肺修复。具体目标 2. 确定 LUBAC 的激活是否介导肺中 NF-κB 的激活我们将确定上皮细胞是否指导单核细胞来源的巨噬细胞的修复表型。这些细胞的炎症表型是由微环境变化驱动的——特别是肺部上皮，或者这些变化是否是巨噬细胞内细胞自主的。具体目标 3. 确定 NF-κB 依赖性炎症基因的表达是否增加重症肺炎患者肺泡巨噬细胞和BAL液与30天相关我们将利用成功临床反应提供的基础设施。肺炎治疗 (SCRIPT) 试验以确定 NF-的表达之间是否存在关联 肺泡巨噬细胞和 BAL 液细胞因子中的 B 靶基因与临床结果。