Expanding Excellence in Developmental Biology in Oklahoma Supplement: 3D Human Lung Tissue Model to Dissect Cellular Responses upon SARS-CoV-2 Infection

俄克拉荷马州增补中扩大发育生物学的卓越性：3D 人体肺组织模型剖析 SARS-CoV-2 感染后的细胞反应

基本信息

批准号：
10853526
负责人：
Linda F Thompson
金额：
$ 64.07万
依托单位：
OKLAHOMA MEDICAL RESEARCH FOUNDATION
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2024-06-30
项目状态：
已结题

项目摘要

Abstract SARS-CoV-2 infection is the causative agent of coronavirus disease 2019 (COVID-19), a global pandemic responsible for >6 million deaths worldwide. The mucociliary epithelium lining the human airways is the primary site of SARS-CoV-2 infection. Following viral exposure, the epithelium initiates responses to recruit immune cells to the site of infection and coordinate the innate immune response. A balance in the number of each cell type (i.e., ciliated, secretory) is critical to maintaining a healthy epithelium, and airway epithelial damage and changes in cell type abundance (termed remodeling) play important roles in the pathology associated with SARS-CoV-2 infection. However, our understanding of how immune cell interactions and emergent SARS-CoV-2 variants impact virus-induced damage and remodeling of the airway epithelium is limited. Our project will test the central hypothesis that myeloid cell responses and interactions with epithelial cells are major drivers of inflammation- mediated epithelial cell remodeling in SARS-CoV-2 infection of the upper airway. Using a novel in vitro 3D model of the human upper airway, composed of primary differentiated airway epithelial cells, lung fibroblasts, pulmonary microvascular endothelial cells and myeloid immune cells, combined with cutting edge scRNA-seq technology, our team will address the following unresolved questions regarding the host immune response to SARS-CoV-2. In Aim 1, we will determine how cross-talk between epithelial and innate myeloid cells in the initial stages of SARS-CoV-2 infection impacts the cell differentiation processes that drive pathological remodeling and damage of the airway epithelium. In Aim 2, we will determine if the SARS-CoV-2 ancestral strain and the Delta and Omicron variants elicit distinct host immune responses and pathogenic epithelial remodeling upon initial infection of the same host environment. The strengths of our novel 3D airway model include the ability to track the kinetics of the host cell response at defined times post-infection to capture early and later events and the ability to compare the impact of each SARS-CoV-2 variant on the same set of donor human cells. To accomplish this project, we have assembled a multi-disciplinary team with non-overlapping and synergistic expertise. Dr. Matthew Walters (Co-Project Lead), a lung cell biologist and virologist, will oversee the SARS-CoV-2 infection studies and characterize the epithelial remodeling phenotypes. Dr. Susan Kovats (Co-Project Lead), an immunologist with expertise in the pulmonary myeloid response to respiratory viruses, will oversee lung model construction and characterize the tissue immune response. Dr. Willard Freeman (Co-Investigator), an expert in transcriptome profiling, will oversee the scRNA-seq analyses. The data collected will form the basis of new multi- PI NIH R01 proposals with the goal to understand how different host environments (e.g., age across the lifespan, gender and chronic lung disease) modulate host immune responses and airway epithelium remodeling upon infection by respiratory viruses such as SARS-CoV-2, RSV and influenza virus.

抽象的 SARS-COV-2感染是2019年冠状病毒病的病因（Covid-19），这是全球大流行病全球造成> 600万人死亡。人类气道的粘膜纤毛上皮是主要的 SARS-COV-2感染部位。病毒暴露后，上皮引发对募集免疫细胞的反应到感染部位并协调先天免疫反应。每种单元格的数量平衡（即纤毛，分泌）对于维持健康的上皮和气道上皮损害和变化至关重要在细胞类型中的丰度（称为重塑）在与SARS-COV-2相关的病理中起重要作用感染。但是，我们对免疫细胞相互作用和新兴SARS-COV-2变体的理解撞击病毒引起的损伤和气道上皮的重塑是有限的。我们的项目将测试中央假设髓样细胞反应以及与上皮细胞相互作用是炎症的主要驱动因素上呼吸道的SARS-COV-2感染中介导的上皮细胞重塑。使用新颖的体外3D模型人类上部气道，由原发性差异气道上皮细胞，肺成纤维细胞，肺部组成微血管内皮细胞和髓样免疫细胞，结合尖端的SCRNA-SEQ技术，我们的团队将解决以下有关主机免疫反应SARS-COV-2的问题。在AIM 1中，我们将确定在初始阶段的上皮和先天髓样细胞之间的串扰 SARS-COV-2感染会影响驱动病理重塑和损害的细胞分化过程气道上皮。在AIM 2中，我们将确定SARS-COV-2祖先应变和三角洲以及 Omicron变体引起不同宿主免疫反应和初始感染后的致病性上皮重塑相同的主机环境。我们新颖的3D气道模型的优势包括跟踪动力学的能力在感染后定义的时间捕获早期和以后的事件的宿主细胞响应的能力比较每个SARS-COV-2变体对同一组供体人细胞的影响。实现这一目标项目，我们组建了一个具有非重叠和协同专业知识的多学科团队。博士肺部细胞生物学家和病毒学家马修·沃尔特斯（Matthew Walters）（共同项目铅）将监督SARS-COV-2感染研究并表征上皮重塑表型。 Susan Kovats博士（共同项目铅），免疫学家在肺髓样反应对呼吸道病毒的反应方面具有专业知识，将监督肺模型结构和表征组织免疫反应。威拉德·弗里曼（Willard Freeman）博士（共同研究者），专家转录组分析将监督SCRNA-SEQ分析。收集到的数据将构成新的多多 PI NIH R01提案的目标是了解不同的主机环境（例如，整个寿命的年龄，性别和慢性肺病）调节宿主免疫反应和气道上皮重塑 SARS-COV-2，RSV和流感病毒等呼吸道病毒感染。