Project 2

项目摘要 在过去的四个月里，SARS冠状病毒-2(SARS-CoV-2)迅速出现，导致了一场关键的 冠状病毒病大流行(新冠肺炎)全球超过140万病例 (https://coronavirus.jhu.edu/map.html)和预计到8月份仅美国就有大约100,000人死亡 2020年(见https://covid19.healthdata.org/projections).SARS-CoV-2会导致致命的急性呼吸窘迫综合征。尽管我们 提高对ARDS的机制认识，临床干预具有挑战性。非-AI-20-31指示 有几个需求，如开发试剂和病毒特征分析，了解关键 病毒感染、复制、发病机制和传播方面的研究、鉴定和评价 对SARS-CoV-2的细胞和体液免疫反应，我们在本提案中解决。 的确，迫切需要了解新冠肺炎的免疫病理机制，研究 肺上皮与组织、免疫系统与病毒的相互作用 这种多方互动。我们需要更好地了解新冠肺炎的免疫病理机制来探索 有可能在新冠肺炎患者中发挥作用的新的治疗方法。 我们的建议从肺上皮细胞对SARS的反应的角度来解决这一需要- CoV-2感染和从T细胞角度来看，新冠肺炎。同时，或者努力也将提供一个 共享的肺呼吸道有机物/SARS-CoV-2/免疫细胞研究平台将加快检测 实验治疗学。我补充计划的结果将与戈登、鲁尼博士分享， 和克鲁梅尔加入我们的‘RapidPath’计划(见支持信)，以促进快速发现和进步 并将与来自新冠肺炎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的IMPACC项目。在这份行政副刊中，我们将利用我的实验室建立的 擅长T细胞信号、T细胞激活、抗原识别、炎症和自身免疫性疾病。 这些都是父母P01(2P01AI091580，Weiss)的广泛话题。独特的是，我们将把我们的T细胞 凭借我们在上皮细胞类有机化合物的生成和研究方面的专业知识。我们已经有了一个 “空中有机体生物库”，我们将把它作为一个社区资源进行扩展。我们将描述 呼吸道对6种不同SARS-CoV-2毒株与H1N1pdm病毒的上皮反应比较 有机体、单细胞RNAseq-和细胞外基质技术(目标1)。为了更好地了解SARS-CoV-2 和获得性免疫，我们将获得对T细胞激活和T细胞信号转导的机械性见解。 SARS-CoV-2和H1N1pdm的背景--七种不同的呼吸道有机物和两种非小细胞肺癌的感染 有机化合物(目标2)。这些“病毒-T细胞-器官”的高分辨率成像和细胞飞行时间分析 提供对SARS-CoV-2及其T细胞生物学亟需的免疫学见解 AI-20-31，并将与“RapidPath”和加州大学旧金山分校IMPACC计划中的其他项目进行协同。

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.