Targeting abnormal alveolar immune activation and failed epithelial repair in COVID-19

针对 COVID-19 中异常的肺泡免疫激活和失败的上皮修复

基本信息

批准号：
10596990
负责人：
GR Scott Budinger
金额：
$ 74.22万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10596990
关键词：
2019-nCoV ACE2 Acute Respiratory Distress Syndrome Address Alveolar Alveolar Macrophages Alveolitis Animal Model Animals Anti-Inflammatory Agents Antibodies Antigen-Presenting Cells Antiviral Agents Attenuated Biological Biological Markers Bronchoalveolar Lavage Fluid COVID-19 COVID-19 patient COVID-19 pneumonia Calcium Calcium Channel Cell Communication Clinical Data Clinical Trials Clinical Trials Design Coronavirus Credentialing Cross Reactions Data Data Science Disease Dose Enrollment Feedback Flow Cytometry Future Generations Goals Heterogeneity Human Immunology In Vitro Infection Inflammasome Inflammation Inflammatory Interleukin-1 beta Interruption Liquid substance Lung Macrophage Measures Mechanical ventilation Mediating Minority Nature Patients Pneumonia Production Public Health Publications Pulmonary Inflammation Research Personnel Role SARS-CoV-2 infection Sampling Secondary to Severity of illness Signal Transduction Sorting T memory cell T-Cell Activation T-Lymphocyte Testing Tissues Virus cross reactivity cytokine direct application env Gene Products epithelial repair genomic data human disease human tissue immune activation inhibitor lymph nodes mutant novel pandemic disease participant enrollment pathogen pharmacologic post SARS-CoV-2 infection predictive modeling severe COVID-19 single-cell RNA sequencing small molecule small molecule inhibitor stem systemic inflammatory response transcriptomic profiling virology

项目摘要

PROJECT SUMMARY This application directly stems from our recent publication in Nature in which we analyzed bronchoalveolar lavage (BAL) fluid from 88 patients with critical SARS-CoV-2 pneumonia using a combination of flow cytometry, bulk transcriptomic profiling of flow sorted alveolar macrophages and, in some patients, single cell RNA- sequencing. We compared these data with analogous samples collected from 211 patients with pneumonia secondary to other pathogens before and during the pandemic with a goal of identifying unique pathobiologic features of SARS-CoV-2 pneumonia. Using these data, we generated the hypothesis that SARS-CoV-2 causes a slowly unfolding, spatially limited alveolitis in which alveolar macrophages harboring SARS-CoV-2 and cross-reactive T cells form a positive feedback loop that drives progressive alveolar inflammation. We address key questions from this hypothesis in three interrelated Specific Aims. Aim 1. To determine whether alveolar macrophage infection and activation of cross-reactive memory T cells drive alveolar macrophage/T cells circuits in patients with SARS-CoV-2 pneumonia. We will examine the role of cross-reactive antibodies in mediating alveolar macrophage infection with SARS-CoV-2. We will also examine the role of cross-reactive memory T cells recognizing SARS-CoV-2 and other coronaviruses in establishing and maintaining positive feedback loops between T cells and infected alveolar macrophages and the role of these signaling loops in initiating persistent lung and systemic inflammation. Aim 2. To determine whether calcium channel activation by the envelope protein of SARS-CoV-2 is necessary for activation of IL-1β in human alveolar macrophages. We will infect human alveolar macrophages with mutant SARS-CoV-2 viruses lacking calcium channel activity in the envelope protein and measure their generation of IL-1β in human alveolar macrophage in vitro. Aim 3. To determine whether a pharmacologic inhibitor of CRAC channel activation can attenuate alveolitis in patients with severe SARS-CoV-2 pneumonia by disrupting circuits between infected alveolar macrophages and cross-reactive T cells. We are actively enrolling in a clinical trial to determine the biologic effects of a small molecule inhibitor of CRAC channel activation using sequential analysis of BAL fluid collected from patients with SARS-CoV-2 pneumonia requiring mechanical ventilation. We will use novel data science approaches to integrate the clinical and genomic data generated from this study. We have assembled a unique group of investigators with expertise in lung immunology, clinical trials, calcium channels and virology. Our studies will explore possible reasons for the observed variability in disease severity after SARS-CoV-2 infection, offer mechanisms to credential CRAC channel inhibitors as both anti-inflammatory and possible antiviral therapeutics in patients with severe SARS-CoV-2 pneumonia, and provide a framework for future clinical trials designed using biomarkers derived from bronchoalveolar lavage fluid.

项目概要该应用直接源于我们最近在《自然》杂志上发表的文章，其中我们分析了支气管肺泡结合流式细胞术对 88 名重症 SARS-CoV-2 肺炎患者的灌洗 (BAL) 液进行分析，对流式分选的肺泡巨噬细胞以及某些患者的单细胞 RNA 进行批量转录组分析我们将这些数据与从 211 名肺炎患者收集的类似样本进行了比较。在大流行之前和期间继发于其他病原体，目的是确定独特的病理生物学利用这些数据，我们得出了 SARS-CoV-2 引起的假设。一种缓慢展开的、空间有限的肺泡炎，其中肺泡巨噬细胞携带 SARS-CoV-2 和交叉反应性 T 细胞形成正反馈回路，驱动进行性肺泡炎症。在三个相互关联的具体目标中解决该假设的关键问题。目的 1. 确定肺泡巨噬细胞是否感染并激活交叉反应记忆 T 我们将检查 SARS-CoV-2 肺炎患者中细胞驱动肺泡巨噬细胞/T 细胞回路的情况。我们还将研究交叉反应抗体在介导肺泡巨噬细胞感染 SARS-CoV-2 中的作用。研究识别 SARS-CoV-2 和其他冠状病毒的交叉反应记忆 T 细胞的作用建立并维持 T 细胞和受感染的肺泡巨噬细胞之间的正反馈循环这些信号环路在引发持续性肺部和全身炎症中的作用。目标 2. 确定 SARS-CoV-2 包膜蛋白对钙通道的激活是否有效人类肺泡巨噬细胞中IL-1β激活所必需的我们将感染人类肺泡。携带突变型 SARS-CoV-2 病毒的巨噬细胞，其包膜蛋白中缺乏钙通道活性，体外测量人肺泡巨噬细胞中 IL-1β 的产生。目标 3. 确定 CRAC 通道激活的药物抑制剂是否可以减弱通过破坏感染者之间的回路来治疗严重 SARS-CoV-2 肺炎患者的肺泡炎我们正在积极参加一项临床试验，以确定肺泡巨噬细胞和交叉反应性 T 细胞。使用 BAL 液序贯分析研究 CRAC 通道激活小分子抑制剂的生物学效应我们将使用从需要机械通气的 SARS-CoV-2 肺炎患者收集的新数据。科学方法整合本研究产生的临床和基因组数据。我们组建了一支独特的研究人员小组，他们在肺免疫学、临床试验、钙我们的研究将探讨观察到的疾病严重程度差异的可能原因。 SARS-CoV-2 感染后，提供了证明 CRAC 通道抑制剂具有抗炎作用的机制以及严重 SARS-CoV-2 肺炎患者可能的抗病毒治疗，并提供一个框架用于使用源自支气管肺泡灌洗液的生物标志物设计的未来临床试验。