Project 2

项目摘要 SARS冠状病毒-2（SARS-CoV-2）在过去四个月迅速出现，导致严重的 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，仅在美国就有大约10万人死亡 2020年（见https：//covid19.healthdata.org/projects）。SARS-CoV-2可导致致命的ARDS。尽管我们 提高对ARDS的机制理解，临床干预具有挑战性。NOT-AI-20-31指示 一些需求，如开发用于病毒表征的试剂和测定法， 病毒感染、复制、发病机制和传播方面， 细胞和体液免疫反应的SARS-CoV-2，我们在这个建议。 事实上，迫切需要了解COVID-19的免疫病理学，并研究COVID-19的免疫机制。 肺上皮和组织，免疫系统和病毒的相互作用，以了解生物学的 这种多方互动。我们需要更好地了解COVID-19的免疫病理学， 新的治疗方法，有可能对COVID-19患者有效。 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感染和COVID-19中的T细胞观点。同时，或努力也将提供一个 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 实验治疗学我的补充计划的结果将与戈登博士，鲁尼博士， 和Krummel在我们的“快速路径”计划（见支持信），以促进快速发现和进展 并将与来自COVID-19患者免疫系统的见解进行比较，同时在 UCSF IMPACC项目。在这份行政补充文件中，我们将利用我的实验室建立的 在T细胞信号传导、T细胞活化、抗原识别、炎症和自身免疫性疾病方面的专业知识。 这些是母公司P01（2 P01 AI 091580，韦斯）的广泛主题。独特的是，我们将联合收割机结合我们的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 “气道类器官生物库”，我们将扩大为社区的资源。我们将描述 使用气道对六种不同SARS-CoV-2毒株与H1N1 pdm病毒的上皮反应进行比较， 类器官、单细胞RNAseq和CyTOF技术（Aim 1）。为了更好地了解SARS-CoV-2 和适应性免疫，我们将获得对T细胞活化和T细胞信号传导的机制见解， SARS-CoV-2和H1N1 pdm感染7种不同的气道类器官和2种NSCLC的背景 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 提供了急需的免疫学见解SARS-CoV-2和它的T细胞生物学，如在非- AI-20-31，并将协同与其他项目在'快速路径'和UCSF IMPACC计划。

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.