Modeling early SARS-CoV-2 pathogenesis in human lung organoids and slice cultures

在人肺类器官和切片培养物中模拟早期 SARS-CoV-2 发病机制

基本信息

批准号：
10449059
负责人：
Arjun Rustagi
金额：
$ 19.33万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10449059
关键词：
2019-nCoV 3-Dimensional Acute Acute respiratory infection Advisory Committees Air Alveolar Cell Alveolar Macrophages Animals Autoantibodies Automobile Driving Basal Cell Biology COVID-19 COVID-19 pandemic COVID-19 pathogenesis COVID-19 patient Cell Line Cells Cessation of life Communicable Diseases Data Dependence Development Development Plans Disease Environment Epithelial Epithelial Cells Event Flow Cytometry Funding Gene Expression Genes Goals Human Immune Immune response Immunity Immunology In Vitro Infection Inflammation Inflammatory Inflammatory Response Innate Immune Response Interferon Type I Interferons Interleukin-18 Interleukin-6 Knowledge Liquid substance Lung Lung diseases Macrophage Activation Manuscripts Maps Mentors Microscopy Modeling Mucous Membrane Mutation Organoids Pathogenesis Pathogenicity Pathway interactions Patients Physicians Play Positioning Attribute Process Protein Analysis RNA amplification Research Research Training Resources Rest Reverse Transcriptase Polymerase Chain Reaction Role SARS-CoV-2 infection SARS-CoV-2 pathogenesis SARS-CoV-2 variant Scientist Signal Transduction Slice Stimulus Structure of parenchyma of lung Supporting Cell System TNF gene Techniques Testing Therapeutic Intervention Time Training Transcript Tumor-infiltrating immune cells Universities Variant Viral Viral Pathogenesis Viral Proteins Virus Virus Replication Work acute infection base career career development cell type combat cytokine design global health high risk immune activation in vivo innate immune pathways insight instructor interest macrophage nasopharyngeal swab novel novel diagnostics novel therapeutic intervention novel therapeutics pandemic disease programs protein expression remdesivir respiratory respiratory pathogen respiratory virus response severe COVID-19 single cell analysis single-cell RNA sequencing statistics stem cells success targeted treatment tool vaccine delivery vaccine hesitancy variants of concern

项目摘要

PROJECT SUMMARY COVID-19 remains an ongoing global health crisis, in part due to emerging variants. Fundamental questions remain about the mechanisms by which SARS-CoV-2 infection drives inflammation in the lower lung, in part due to the lack of in vitro infection models. Better understanding of variant biology and the immune pathways involved in early SARS-CoV-2 infection could offer insights for the development of novel therapeutic strategies. The antiviral cytokines type I interferons and their downstream effects have been implicated in COVID-19. SARS-CoV-2 may inhibit or promote interferon effects in some cell types in the lung, and this may differ between viral variants. My preliminary work shows that organoids and lung slice cultures can be used to study these early events in SARS-CoV-2 infection and identifies macrophages as a cell type that activates the interferon pathway in infected cultures. Thus, I propose using quantitative PCR, single-cell RNA sequencing, and flow cytometry to study viral variants and to deeply profile the changes to viral and host gene and protein expression during infection. I will study highly relevant cell types that can interact with each other in a similar fashion to the in vivo lung. I will study the interferon stimulated gene response and identify the cell types in which it is being modulated or would be good targets for therapeutic intervention. This knowledge will be critical to our efforts to combat the COVID-19 pandemic. I am currently a fellow in Dr. Catherine Blish’s lab in the Division of Infectious Diseases at Stanford University and I am in the process of being promoted to a full time Instructor position. Stanford University offers an outstanding scientific environment, where all the resources necessary to the success of this project are made available to me. My long-term goal is to become an independent physician-scientist, with a research focus in respiratory viral pathogenesis. I aim to establish a research program focused on modeling established and emerging infections in primary lung tissue with a goal of developing new treatments. To achieve this goal, I have designed a tailored career development plan that comprises both formal and informal training. This training will enhance my expertise in lung biology, immunology, and sequencing analysis. Informal training in these topics will be provided by my mentor, Dr. Catherine Blish, and by the rest of my advisory committee, Dr. Calvin Kuo, Dr. Mark Krasnow, Dr. Susan Holmes, and Dr. Ben Pinsky. With their guidance, I will complete the proposed project, submit research manuscripts, obtain further funding, and obtain an independent research position.

项目概要 COVID-19 仍然是一场持续的全球健康危机，部分原因是新出现的基本问题。部分仍然是关于 SARS-CoV-2 感染炎症在下肺中驱动的机制由于缺乏体外感染模型，无法更好地了解变异生物学和免疫途径。参与早期 SARS-CoV-2 感染可以为开发新的治疗策略提供见解。抗病毒细胞因子 I 型干扰素及其下游效应与 COVID-19 有关。 SARS-CoV-2 可能抑制或促进肺部某些细胞类型的干扰素作用，这可能有所不同我的初步工作表明类器官和肺切片培养物可用于研究这些 SARS-CoV-2 感染的早期事件将巨噬细胞确定为激活因此，我建议使用定量 PCR、单细胞 RNA 测序，和流式细胞术来研究病毒变异并深入分析病毒和宿主基因和蛋白质的变化我将研究可以以类似方式相互作用的高度相关的细胞类型。我将研究干扰素刺激的基因反应并鉴定其中的细胞类型。它正在被调节或将成为治疗干预的良好目标。这一知识将至关重要。来支持我们抗击 COVID-19 大流行的努力。我目前是斯坦福大学传染病系 Catherine Blish 博士实验室的研究员我正在晋升为全职讲师职位，斯坦福大学提供了一个职位。优秀的科学环境，为该项目的成功提供了所有必要的资源我的长期目标是成为一名独立的医师科学家，研究重点是我的目标是建立一个研究计划，重点是建立模型和原发性肺组织中新出现的感染，旨在开发新的治疗方法。为了实现这一目标，我设计了一份量身定制的职业发展计划，包括正式和该培训将增强我在肺生物学、免疫学和测序方面的专业知识。这些主题的非正式培训将由我的导师凯瑟琳·比利什博士和其他人提供。我的顾问委员会成员包括 Calvin Kuo 博士、Mark Krasnow 博士、Susan Holmes 博士和 Ben Pinsky 博士。指导，我将完成拟议的项目，提交研究手稿，获得进一步的资助，并获得一个独立的研究职位。