Project 2

项目2

• 批准号: 10208652
• 负责人: JEROEN ROOSE
• 金额: $ 40.53万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-18 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10208652
• 关键词: 2019-nCoV; Address; Administrative Supplement; Adult Respiratory Distress Syndrome; Autoimmune Diseases; Biochemistry; Biological Assay; Biology; CD27 Antigens; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; COVID-19; Cancer Patient; Carcinoma; Cell Communication; Cell Culture Techniques; Cells; Cellular biology; Clinical; Collection; Communities; Complex; Containment; Coronavirus; Data Science; Data Set; Development; Disease; Epithelial; Epithelial Cells; Epithelium; Evaluation; Gene Expression; Generations; Genomics; Goals; Human; Immune; Immune response; Immune system; Immunologics; Infection; Inflammation; Influenza; Influenza A Virus, H1N1 Subtype; Interferons; Intervention; Investigational Therapies; Knowledge; Letters; Libraries; Link; Lung; Lung infections; Maps; Non-Small-Cell Lung Carcinoma; Organoids; Parents; Pathogenesis; Pathologic; Patients; Play; Production; Property; Reagent; Research; Resources; Role; Signal Transduction; Specialist; Stains; T Virus; T cell response; T-Cell Activation; T-Lymphocyte; Technology; Testing; Tissues; Viral; Virus; Virus Diseases; Work; adaptive immunity; biobank; biosafety level 3 facility; computerized data processing; coronavirus disease; cytotoxic CD8 T cells; high resolution imaging; immune function; immunopathology; improved; influenzavirus; insight; novel therapeutic intervention; pandemic disease; programs; response; sample fixation; single-cell RNA sequencing; transmission process

PROJECT ABSTRACT The SARS coronavirus-2 (SARS-CoV-2) has rapidly emerged over the past four months leading to a critical pandemic of coronavirus disease (COVID-19) with over 1.4M cases worldwide (https://coronavirus.jhu.edu/map.html) and roughly 100,000 projected fatalities in the US alone by August 2020 (See https://covid19.healthdata.org/projections). SARS-CoV-2 causes a lethal ARDS. Despite our improved mechanistic understanding of ARDS, intervention clinically is challenging. NOT-AI-20-31 indicated several needs, such as development of reagents and assays for virus characterization, understand critical aspects of viral infection, replication, pathogenesis, and transmission, identification and evaluation of the cellular and humoral immune responses to SARS-CoV-2, which we address in this proposal. Indeed, there is an urgent need to understand the immunopathology of COVID-19 and study the interactions of the lung epithelium and tissue, the immune system and the virus to understand the biology of this multipartite interaction. We need to better understand the immunopathology of COVID-19 to explore novel therapeutic approaches that have the potential to work in COVID-19 patients. Our proposal addresses this need from the perspective of the lung epithelium response to SARS- CoV-2 infection and from a T cell perspective in COVID-19. Simultaneously, or efforts will also provide a sharable research platform of lung airway organoids/SARS-CoV-2/immune cells that will expedite testing of experimental therapeutics. Results from my supplement program will be shared with Drs. Gordon, Looney, and Krummel in our ‘RapidPath’ program (see supporting letter) to promote rapid discovery and progress and will be compared to insights from COVID-19 patient immune systems, being simultaneously profiled in the UCSF IMPACC project. In this this Administrative Supplement we will capitalize on my lab’s established expertise in T cell signaling, T cell activation, antigen recognition, inflammation, and autoimmune diseases. Those are broad topics of the parent P01 (2P01AI091580, Weiss). Uniquely, we will combine our T cell expertise with our expertise in the generation and studies of epithelial cell organoids. We already have an “Airway Organoid Biobank” that we will expand as a resource for the community. We will characterize the epithelial response to six different SARS-CoV-2 strains compared to H1N1pdm virus, using airway organoid-, single cell RNAseq-, and CyTOF- technology (Aim 1). In order to better understand SARS-CoV-2 and adaptive immunity, we will obtain mechanistic insights into T cell activation and T cell signaling in the context of SARS-CoV-2- and H1N1pdm- infection of seven, diverse Airway Organoids and two NSCLC organoids (Aim 2). High-resolution imaging and CyTOF analysis of these “virus-T cell-organoids” will provide much needed immunological insights into SARS-CoV-2 and its T cell biology as indicated in NOT- AI-20-31 and will synergize with other projects in ‘RapidPath’ and in UCSF IMPACC programs.

项目摘要 SARS 冠状病毒-2 (SARS-CoV-2) 在过去四个月中迅速出现，导致了一场严重的疫情 冠状病毒病 (COVID-19) 大流行，全球病例超过 140 万例 (https://coronavirus.jhu.edu/map.html)，预计到 8 月份，仅美国就有大约 100,000 人死亡 2020 年（参见 https://covid19.healthdata.org/projections）。 SARS-CoV-2 会导致致命的 ARDS。尽管我们的 尽管提高了对 ARDS 机制的认识，但临床干预仍具有挑战性。 NOT-AI-20-31 已指示 一些需求，例如开发病毒表征的试剂和分析方法，了解关键 病毒感染、复制、发病机制和传播等方面，以及病毒的识别和评估 我们在本提案中讨论了针对 SARS-CoV-2 的细胞和体液免疫反应。 事实上，迫切需要了解 COVID-19 的免疫病理学并研究 肺上皮和组织、免疫系统和病毒的相互作用，以了解肺上皮和组织、免疫系统和病毒的生物学 这种多方互动。我们需要更好地了解 COVID-19 的免疫病理学来探索 可能对 COVID-19 患者有效的新型治疗方法。 我们的建议从肺上皮对 SARS 反应的角度解决了这一需求 CoV-2 感染以及从 T 细胞的角度来看 COVID-19。同时，或者努力也将提供一个 肺气道类器官/SARS-CoV-2/免疫细胞的共享研究平台，将加快测试 实验疗法。我的补充计划的结果将与博士分享。戈登、卢尼、 和 Krummel 参与我们的“RapidPath”计划（参见支持信），以促进快速发现和进步 并将与来自 COVID-19 患者免疫系统的见解进行比较，同时在 加州大学旧金山分校 IMPACC 项目。在本行政补充文件中，我们将利用我的实验室已建立的 T 细胞信号传导、T 细胞激活、抗原识别、炎症和自身免疫性疾病方面的专业知识。 这些是父 P01 (2P01AI091580, Weiss) 的广泛主题。独特的是，我们将结合我们的 T 细胞 我们在上皮细胞类器官的生成和研究方面的专业知识。我们已经有一个 我们将扩展“气道类器官生物库”作为社区的资源。我们将表征 与 H1N1pdm 病毒相比，使用气道对六种不同 SARS-CoV-2 毒株的上皮反应 类器官、单细胞 RNAseq 和 CyTOF 技术（目标 1）。为了更好地了解 SARS-CoV-2 和适应性免疫，我们将获得有关 T 细胞激活和 T 细胞信号传导的机制见解 七种不同气道类器官和两种非小细胞肺癌 (NSCLC) 的 SARS-CoV-2- 和 H1N1pdm- 感染背景 类器官（目标 2）。这些“病毒-T细胞-类器官”的高分辨率成像和 CyTOF 分析将 为 SARS-CoV-2 及其 T 细胞生物学提供急需的免疫学见解，如 NOT- 中所示 AI-20-31 并将与“RapidPath”和 UCSF IMPACC 项目中的其他项目产生协同作用。