COVID-19 imprints airway basal cells to impair epithelium regeneration

COVID-19 印记气道基底细胞，损害上皮再生

• 批准号: 10738549
• 负责人: Xingbin Ai
• 金额: $ 26.43万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-25 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738549
• 关键词: 2019-nCoV; ATAC-seq; Acute respiratory infection; Age; Air; Animal Model; Antibodies; Basal Cell; Biological Assay; Biopsy; COVID-19; COVID-19 impact; Cell Cycle Arrest; Cell Line; Cell Separation; Cell Survival; Cell model; Cells; Cessation of life; Chromatin; Chronic lung disease; Clinical Research; Contracts; Coronavirus; Defect; Derivation procedure; Disease; EZH2 gene; Endowment; Epigenetic Process; Epithelium; Exhibits; Foundations; Functional disorder; Future; Gene Expression; Genetic study; Goals; Health; Human; Hyperactivity; Immune; Impairment; In Vitro; Infection; Inflammation; Inflammation Mediators; Inflammatory; Intubation; Lead; Liquid substance; Lower respiratory tract structure; Lung; Lung diseases; Mediating; Mediator; Medical Waste; Memory; Methods; Microscopic; Modeling; Modification; Molecular; Morbidity - disease rate; Mus; Natural Immunity; Olfactory Pathways; Organ; Pathogenesis; Pathway interactions; Patient Care; Patients; Phenotype; Play; Procedures; Proliferating; Research; Respiratory Disease; Respiratory Failure; Respiratory Tract Infections; Risk; Risk Reduction; Role; SARS-CoV-2 infection; SARS-CoV-2 negative; STAT3 gene; Sampling; Signal Transduction; Site; Skin; Source; Spanish flu; Stains; Testing; Therapeutic; Trachea; United States; Viral Respiratory Tract Infection; Virus; acute infection; airway epithelium; alveolar type II cell; aspirate; chemokine; cytokine; epigenetic memory; epithelial stem cell; epithelium regeneration; genetic approach; genetic signature; genomic locus; imprint; in vivo; mortality; multimodality; pandemic disease; pandemic influenza; pathogen; prevent; programs; receptor; regeneration function; respiratory; response; secondary infection; senescence; stem cell proliferation; stem cells; stem-like cell; therapeutic target; transcription factor; transcriptome

PROJECT SUMMARY COVID-19 (CoV19) is caused by SARS-CoV-2 infection of the airway epithelium resulting in extensive damage in the lower respiratory tract. Since 2020, CoV19 has claimed over 1 million lives in the United States surpassing the death toll of the 1918 H1N1 influenza pandemic. Clinical studies show that the mortality and morbidity of CoV19 is associated with secondary infection. Given a central role of the airway epithelium as a barrier against pathogens, preliminary studies tested whether airway basal stem cells (BSCs) are impaired in their regenerative function in CoV19 patients. Of note, BSCs are a major type of stem cells responsible for epithelium regeneration following respiratory viral infection in humans. In preliminary studies, we derived 6 lines of BSCs from severe cases of CoV19 using tracheal aspirate (TA) as a source of bronchial BSCs. These CoV19-exposed BSCs were tested free of virus; however, they show early cell cycle arrest, sustained STAT3 hyperactivity, and defective differentiation in air-liquid interface. In contract, BSCs derived from TA of control patients with neurogenic and cardiogenic respiratory failure have no such defects. Similar to our findings in vitro, antibody staining of fatal CoV19 lung sections revealed increased senescence and defective differentiation of BSCs. In addition, CoV19- exposed BSCs in vitro maintained, at least partially, an inflammatory gene signature that was found in BSCs in vivo by single cell-seq of lung samples from CoV19 patients. As such, BSCs derived from TA of CoV19 patients provide a viable cell model to investigate how CoV19 impairs epithelial regeneration by inducing an inflammatory memory in BSCs. Mechanistically, CoV19-exposed BSCs exhibit unique chromatin opening at sites enriched for transcriptional factors mediating the inflammatory pathways, such as STAT3. Based on these preliminary findings, we hypothesize that inflammation in CoV19 causes an epigenetic memory in BSCs to impair epithelium regeneration. Leveraging our ability to derive TA BSCs, Aim1 will test whether CoV19 uniquely reprograms BSCs compared to other acute respiratory infections. Aim 2 will identify the molecular mediators of the inflammatory memory in CoV19-exposed BSCs using complementary assays. The rescue assay will test whether blocking STAT3 hyperactivity and reversing epigenetic modification in CoV19-exposed BSCs will normalize their role in epithelial regeneration. The disease-mimicking assay will assess the activity of inflammatory signals in memory induction in healthy control BSCs. The proposed exploratory studies will lay the foundation for future delineation of inflammatory signals and intracellular mediators in the disease memory of BSCs using genetic approaches and animal models of SARS-CoV-2 infection. Our findings will inform therapeutics to facilitate epithelial regeneration in severe cases of CoV19.

项目摘要 COVID-19（CoV19）是由SARS-CoV-2感染气道上皮引起的， 下呼吸道损伤自2020年以来，CoV19已在美国夺去了超过100万人的生命 超过1918年H1N1流感大流行死亡人数的州。临床研究表明， CoV19的死亡率和发病率与继发感染有关。由于气道的中心作用 上皮作为抵抗病原体的屏障，初步研究测试了气道基底干细胞是否 在CoV19患者中，BSCs的再生功能受损。值得注意的是，BSC是一种主要类型的 人类呼吸道病毒感染后负责上皮再生的干细胞。在 在初步研究中，我们使用气管抽吸物（TA）从CoV19的严重病例中获得了6个BSC系。 作为支气管BSC的来源。这些暴露于CoV19的BSC经测试不含病毒;然而， 显示早期细胞周期停滞、持续的STAT3过度活性和在空气-液体中有缺陷的分化 接口.相反，来自对照组神经源性和心源性TA的BSC 呼吸衰竭没有这些缺陷。与我们在体外的发现相似，致死性CoV19的抗体染色 肺切片显示BSCs的衰老增加和分化缺陷。此外，CoV19- 体外暴露的BSCs至少部分地维持了一种炎症基因特征， 通过来自CoV19患者的肺样品的单细胞测序的体内BSC。因此，衍生自TA的BSC CoV19患者提供了一个活细胞模型，以研究CoV19如何通过以下方式损害上皮再生： 在BSC中诱导炎症记忆。从机制上讲，CoV19暴露的BSC表现出独特的 染色质在富含介导炎症途径的转录因子的位点开放， 作为STAT 3。基于这些初步发现，我们假设CoV19中的炎症导致了 BSC中的表观遗传记忆损害上皮再生。利用我们的能力， TA BSC，Aim1将测试与其他急性呼吸道疾病相比，CoV19是否独特地重编程BSC 感染.目的2将鉴定暴露于CoV 19的炎症记忆的分子介质 使用互补测定的BSC。拯救测定将测试阻断STAT3活动过度是否 逆转CoV19暴露的BSC中的表观遗传修饰将使它们在上皮细胞中的作用正常化。 再生疾病模拟试验将评估记忆中炎症信号的活性 在健康对照BSC中诱导。建议的探索性研究将为未来的研究奠定基础。 描述炎症信号和细胞内介质在疾病记忆的BSC使用 SARS-CoV-2感染的遗传学方法和动物模型。我们的发现将为治疗提供信息， 促进CoV19严重病例的上皮再生。