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：
10557881
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/10557881
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 Communication Cell Line Cells Cessation of life Communicable Diseases Data Dependence Development Development Plans Disease Environment Epithelial Cells Epithelium Event Flow Cytometry Funding Gene Expression Genes Goals Human IL18 gene Immune Immune response Immunity Immunology In Vitro Infection Inflammation Inflammatory Inflammatory Response Innate Immune Response Interferon Type I Interferons Interleukin-6 Knowledge Liquid substance Lung Lung diseases Macrophage Macrophage Activation Manuscripts Maps Mentors Microscopy Modeling Mucous Membrane Mutation Organoids Pathogenesis Pathogenicity Pathway interactions Patients Physicians Play Positioning Attribute Process Productivity Protein Analysis Quantitative Reverse Transcriptase PCR RNA amplification Research Resources Rest Role SARS-CoV-2 infection SARS-CoV-2 pathogenesis SARS-CoV-2 variant Sampling Scientist Signal Transduction Slice Stimulus Structure of parenchyma of lung Supporting Cell System TNF gene Techniques Testing Therapeutic Intervention Training Transcript Tropism Universities Variant Viral Viral Pathogenesis Viral Proteins Virus Virus Replication Work acute infection career career development cell type combat cytokine design global health high risk immune activation immune cell infiltrate in vivo innate immune pathways insight instructor 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 variant of interest 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.

项目摘要 Covid-19仍然是一场持续的全球健康危机，部分原因是新兴的变体。基本问题仍然围绕SARS-COV-2感染驱动下肺注射的机制，部分由于缺乏体外感染模型。更好地了解变异生物学和免疫途径参与早期SARS-COV-2感染可以为开发新型治疗策略提供见解。 I型抗病毒细胞因子干扰物及其下游效应已在Covid-19中暗示。 SARS-COV-2可能会抑制或促进肺中某些细胞类型的干扰效应，这可能有所不同在病毒变体之间。我的初步工作表明，可以使用器官和肺切片培养物来研究 SARS-COV-2感染中的这些早期事件，并将巨噬细胞鉴定为激活的细胞类型感染培养物中的干扰素途径。那我建议使用定量PCR，单细胞RNA测序，和流式细胞仪研究病毒变体，并深入介绍病毒和宿主基因和蛋白质的变化感染期间的表达。我将研究高度相关的细胞类型，这些细胞类型可以在类似的体内肺的时尚。我将研究干扰素刺激的基因反应，并确定细胞类型它正在调制或将成为治疗干预的良好目标。这些知识将是至关重要的为了打击Covid-19的大流行而努力。我目前是凯瑟琳·布莱什（Catherine Blish）博士在斯坦福大学传染病科的研究员而且我正在晋升为全职讲师职位。斯坦福大学提供杰出的科学环境，该项目成功所需的所有资源我可用。我的长期目标是成为独立的身体科学家，并以研究重点呼吸病毒发病机理。我的目的是建立一个研究计划，旨在建模建立和原发性肺组织中的新兴感染，目的是开发新疗法。为了实现这一目标，我设计了一个量身定制的职业发展计划，包括正式和非正式培训。这种培训将增强我在肺部生物学，免疫学和测序方面的专业知识分析。这些主题的非正式培训将由我的心理凯瑟琳·布莱什（Catherine Blish）博士以及其余部分提供我的咨询委员会，Calvin Kuo博士，Mark Krasnow博士，Susan Holmes博士和Ben Pinsky博士。与他们指导，我将完成拟议的项目，提交研究手稿，获得进一步的资金并获得独立的研究职位。