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

• 批准号: 10696965
• 负责人: GR Scott Budinger
• 金额: $ 53.35万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 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/10696965
• 关键词: 2019-nCoV; Acceleration; Acute; Acute Respiratory Distress Syndrome; Alveolar; Alveolar Macrophages; Attenuated; Biological; Cell physiology; Cells; Cessation of life; Clinical; Complex; Data; Development; Epithelial Cells; Epithelium; 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; Macrophage; Measures; Mediating; Molecular; Morbidity - disease rate; 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; Viral Pneumonia; Virus Diseases; alveolar epithelium; community acquired pneumonia; cytokine; epithelial repair; experimental study; improved outcome; individualized medicine; insight; lung injury; lung repair; monocyte; mortality; mouse model; participant enrollment; pathogen; pneumonia treatment; prevent; repair function; repaired; response; transcription factor; ubiquitin ligase; ubiquitin-protein ligase

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，所有项目和核心都认为呼吸衰竭的持续存在和发展 ARDS患者的多个器官功能障碍是炎症正常机制的失败 和肺修复。该假设在临床上通过对ARDSNET入选的患者的最新分析得到了支持 ARDS的“透明质炎”内型与包括死亡在内的临床结局不佳有关。 编码炎症细胞因子的许多基因的表达受转录因子NF-B的调节。 因此，了解如何在肺部受伤和维修过程中控制NF-B活动如何提供 受伤后肺修复成功或失败的病理生物学的机械洞察力。 线性泛素组装复合物（LUBAC）是一种E3泛素连接酶，执行Met-1泛素化 NF-B激活所必需的。在初步实验中，我们观察到上皮的特异性缺失 LUBAC（HOIL-1L）的调节成分可防止NF-B信号传导并改善鼠的预后 影响Za病毒感染的模型。我们提供了支持我们假设的其他初步数据 LUBAC促进肺泡上皮和单核细胞衍生的肺泡中持续的NF-B信号传导 巨噬细胞抑制影响肺诱导的肺损伤后的肺修复。 具体目的1。确定是否抑制肺上皮中的lubac介导的NF-B激活 在从影响力恢复期间，感染加速了肺修复。我们将执行定时实验 我们将通常抑制LUBAC介导的NF-B信号传导，从影响力A恢复和 使用完整的生理和分子技术来测量肺修复。 具体目的2。确定肺中Lubac介导的NF-B激活是否激活 上皮指导单核细胞衍生的巨噬细胞中的修复表型。我们将确定是否 这些细胞的炎症表型是由不断变化的微环境驱动的，特别是肺 上皮，或这些变化是否是巨噬细胞内的细胞自主。 具体目的3。确定NF-B依赖性炎症基因的表达是否增加 严重肺炎患者的肺泡巨噬细胞和BAL液与30天有关 死亡。我们将利用成功的临床反应提供的基础设施 肺炎治疗（脚本）试验，以确定NF-表达之间是否存在关联 B型肺泡巨噬细胞和BAL流体细胞因子具有临床结局的靶基因。