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

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/10853526
• 关键词: 2019-nCoV; 3-Dimensional; Adult; Age; Air; Alveolar; Antiviral Response; Architecture; Autopsy; Biological Assay; Blood; COVID-19; COVID-19 pandemic; COVID-19 patient; COVID-19 severity; Cell Communication; Cell Differentiation process; Cell Separation; Cells; Cessation of life; Chronic lung disease; Collagen; Data; Dendritic Cells; Development; Developmental Biology; Disease; Dissection; Elderly; Endothelium; Environment; Epithelial Cells; Epithelium; Equilibrium; Event; Fibroblasts; Gender; Gene Expression; Genetic Transcription; Goals; Histologic; Host Defense; Human; Hydrogels; Immune; Immune response; Immunity; Immunologist; Impairment; In Vitro; Individual; Infection; Infiltration; Inflammation; Influenza A virus; Innate Immune Response; Interferons; Kinetics; Lead; Liquid substance; Longevity; Lung; Macrophage; Measurement; Measures; Mediating; Modeling; Molecular; Myelogenous; Myeloid Cell Activation; Myeloid Cells; Normal Cell; Oklahoma; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Phenotype; Play; Population; Process; Research Personnel; Respiratory Syncytial Virus Infections; Respiratory syncytial virus; Role; SARS-CoV-2 B.1.1.529; SARS-CoV-2 B.1.617.2; SARS-CoV-2 infection; SARS-CoV-2 variant; Science; Severities; Signal Transduction; Site; Structure of parenchyma of lung; Technology; Testing; Time; Tissue Model; Tissues; United States National Institutes of Health; Variant; Viral; Viral Load result; Virus; Virus Diseases; Work; airway epithelium; airway remodeling; cell type; chemokine; cytokine; immune cell infiltrate; in vivo; influenzavirus; interstitial; lung injury; lung microvascular endothelial cells; migration; model building; monocyte; multidisciplinary; novel; pandemic disease; post SARS-CoV-2 infection; recruit; respiratory infection virus; respiratory virus; response; severe COVID-19; single-cell RNA sequencing; three-dimensional modeling; transcriptomic profiling; virus host interaction

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 感染是全球大流行的 2019 年冠状病毒病 (COVID-19) 的病原体 导致全球超过 600 万人死亡。人体呼吸道内壁的粘液纤毛上皮是主要的 SARS-CoV-2 感染部位。病毒暴露后，上皮细胞启动反应以招募免疫细胞 到达感染部位并协调先天免疫反应。每种细胞类型的数量平衡 （即纤毛、分泌）对于维持健康的上皮以及气道上皮的损伤和变化至关重要 细胞类型丰度（称为重塑）在 SARS-CoV-2 相关病理学中发挥重要作用 感染。然而，我们对免疫细胞如何相互作用和新出现的 SARS-CoV-2 变体的理解 病毒引起的气道上皮损伤和重塑的影响是有限的。我们的项目将测试中央 假设骨髓细胞反应和与上皮细胞的相互作用是炎症的主要驱动因素 介导上呼吸道 SARS-CoV-2 感染中的上皮细胞重塑。使用新颖的体外 3D 模型 人上呼吸道的细胞，由初级分化的气道上皮细胞、肺成纤维细胞、肺成纤维细胞组成 微血管内皮细胞和骨髓免疫细胞，结合尖端的scRNA-seq技术， 我们的团队将解决以下有关宿主对 SARS-CoV-2 免疫反应的未解决问题。 在目标 1 中，我们将确定在初始阶段上皮细胞和先天骨髓细胞之间的串扰是如何进行的。 SARS-CoV-2 感染影响细胞分化过程，从而驱动病理重塑和损伤 气道上皮。在目标 2 中，我们将确定 SARS-CoV-2 祖先毒株以及 Delta 和 Omicron 变异在初次感染时引发不同的宿主免疫反应和致病性上皮重塑 同一主机环境。我们新颖的 3D 气道模型的优势包括能够跟踪动力学 感染后特定时间的宿主细胞反应以捕获早期和晚期事件以及捕获早期和晚期事件的能力 比较每种 SARS-CoV-2 变体对同一组供体人类细胞的影响。为了实现这一点 项目中，我们组建了一支具有不重叠和协同专业知识的多学科团队。博士。 肺细胞生物学家和病毒学家 Matthew Walters（联合项目负责人）将负责监督 SARS-CoV-2 感染 研究并表征上皮重塑表型。 Susan Kovats 博士（联合项目负责人） 具有呼吸道病毒肺髓反应专业知识的免疫学家将监督肺模型 构建并表征组织免疫反应。 Willard Freeman 博士（联合研究员），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方面的基础 PI NIH R01 提案的目标是了解不同的宿主环境（例如，整个生命周期中的年龄、 性别和慢性肺病）调节宿主免疫反应和气道上皮重塑 呼吸道病毒感染，如 SARS-CoV-2、RSV 和流感病毒。

项目成果

PDGFRα signaling drives adipose tissue fibrosis by targeting progenitor cell plasticity.

• DOI: 10.1101/gad.260554.115
• 发表时间: 2015-06-01
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Iwayama T;Steele C;Yao L;Dozmorov MG;Karamichos D;Wren JD;Olson LE
• 通讯作者: Olson LE

GTSE1 regulates spindle microtubule dynamics to control Aurora B kinase and Kif4A chromokinesin on chromosome arms.

• DOI: 10.1083/jcb.201610012
• 发表时间: 2017-10-02
• 期刊: The Journal of cell biology
• 作者: Tipton AR;Wren JD;Daum JR;Siefert JC;Gorbsky GJ
• 通讯作者: Gorbsky GJ

AutoGDC: A Python Package for DNA Methylation and Transcription Meta-Analyses.

AutoGDC：用于 DNA 甲基化和转录荟萃分析的 Python 包。

• DOI: 10.1101/2024.04.14.589445
• 发表时间: 2024
• 期刊: bioRxiv : the preprint server for biology
• 作者: Brown,ChaseAlan;Wren,JonathanD
• 通讯作者: Wren,JonathanD

Discovery of a Benzamide Derivative That Protects Pancreatic β-Cells against Endoplasmic Reticulum Stress.

• DOI: 10.1021/acs.jmedchem.7b00435
• 发表时间: 2017-07-27
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Duan H;Li Y;Arora D;Xu D;Lim HY;Wang W
• 通讯作者: Wang W

Variants at multiple loci implicated in both innate and adaptive immune responses are associated with Sjögren's syndrome.

• DOI: 10.1038/ng.2792
• 发表时间: 2013-11
• 期刊: NATURE GENETICS
• 影响因子: 30.8
• 作者: Lessard, Christopher J.;Li, He;Adrianto, Indra;Ice, John A.;Rasmussen, Astrid;Grundahl, Kiely M.;Kelly, Jennifer A.;Dozmorov, Mikhail G.;Miceli-Richard, Corinne;Bowman, Simon;Lester, Sue;Eriksson, Per;Eloranta, Maija-Leena;Brun, Johan G.;Goransson, Lasse G.;Harboe, Erna;Guthridge, Joel M.;Kaufman, Kenneth M.;Kvarnstrom, Marika;Jazebi, Helmi;Graham, Deborah S. Cunninghame;Grandits, Martha E.;Nazmul-Hossain, Abu N. M.;Patel, Ketan;Adler, Adam J.;Maier-Moore, Jacen S.;Farris, A. Darise;Brennan, Michael T.;Lessard, James A.;Chodosh, James;Gopalakrishnan, Rajaram;Hefner, Kimberly S.;Houston, Glen D.;Huang, Andrew J. W.;Hughes, Pamela J.;Lewis, David M.;Radfar, Lida;Rohrer, Michael D.;Stone, Donald U.;Wren, Jonathan D.;Vyse, Timothy J.;Gaffney, Patrick M.;James, Judith A.;Omdal, Roald;Wahren-Herlenius, Marie;Illei, Gabor G.;Witte, Torsten;Jonsson, Roland;Rischmueller, Maureen;Ronnblom, Lars;Nordmark, Gunnel;Wan-Fai Ng;Mariette, Xavier;Anaya, Juan-Manuel;Rhodus, Nelson L.;Segal, Barbara M.;Scofield, R. Hal;Montgomery, Courtney G.;Harley, John B.;Sivils, Kathy L.
• 通讯作者: Sivils, Kathy L.