Live imaging of SARS-CoV-2 infection in novel humanized mice

新型人源化小鼠中 SARS-CoV-2 感染的实时成像

基本信息

批准号：
10400392
负责人：
Michael Allen Brehm
金额：
$ 50万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-15 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10400392
关键词：
2019-nCoV ACE2 Administrative Supplement Animal Model Biology CD34 gene COVID-19 COVID-19 pandemic Cell Death Communities Community Health Cytokeratin 18 Disease Engraftment Foundations Genes Genome Goals Health Human Image Imaging technology Immune Immune response Immune system Immunodeficient Mouse Infection Inflammation Inflammatory Innate Immune Response Innate Immune System Investigation K-18 conjugate Kinetics Knock-in Knowledge Laboratories Metabolic Pathway Modeling Molecular Monitor Mouse Strains Mus Pathogenesis Pathology Physiological Positioning Attribute Protocols documentation Regulation Reporter Research Personnel Resources Role SARS-CoV-2 infection SARS-CoV-2 pathogenesis SARS-CoV-2 variant Systems Development Testing The Jackson Laboratory Time Tissues Transgenic Organisms Transplantation Universities Variant Viral Viral Load result Viral Pathogenesis Virus Virus Diseases cytokine cytokine release syndrome experience global health humanized mouse imaging approach immune activation in vivo in vivo bioluminescence imaging in vivo imaging mouse model nanoluciferase novel pathogenic virus pre-clinical promoter response time use

项目摘要

PROJECT SUMMARY The SARS-CoV-2 pandemic is an unprecedented challenge for the global health community, and there is urgent need to understand the pathogenesis of SARS-CoV-2 and emerging viral variants. The pathology associated with SARS-CoV-2 infection ranges from asymptomatic to severe, in some cases fatal disease that correlates with inflammatory cell death during infection, activation of the immune system and release of inflammatory cytokines or a “cytokine storm”. Unfortunately, available animal models of SARS-CoV-2 infection do not fully recapitulate the human immune response to viral infection due to multiple differences in genomes, molecular and metabolic pathways, and immune system regulation. The goal of our supplemental proposal in response to PA-20-272 is use a panel of translational humanized mouse models developed by Dr. Leonard Shultz at The Jackson Laboratory to study the clearance of SARS-CoV-2 and the variants that are continually emerging, and for the testing of human-specific therapies that will mitigate the pathology associated with COVID-19. In addition, we will use a novel in vivo bioluminescence imaging approach to monitor the kinetics of SARS-CoV-2 spread and clearance in real time in vivo. We have assembled a team of investigators with extensive experience in 1) creating new humanized mouse strains (Dr. Leonard Shultz at The Jackson Laboratory), 2) engrafting human immune systems into immunodeficient mice (Drs. Dale Greiner and Michael Brehm at UMass), 3) the study of human viral pathogens (Dr. Priti Kumar at Yale University), and 4) expertise in imaging virus infection in real time in vivo (Dr. Pradeep Uchil at Yale University). We will leverage the resources of The Jackson Laboratory to facilitate the rapid distribution of effective models to the scientific community, uniquely positioning our group to contribute quickly to the knowledge of SARS-CoV-2 biology. Dr. Shultz has generated new models of NSG mice expressing human ACE2 and will provide these mice to Dr. Kumar and to Drs. Brehm and Greiner. Drs. Brehm and Greiner will engraft the mice with human UCB CD34+ HSCs and will provide engrafted mice to Dr. Kumar’s laboratory. Drs. Kumar and Uchil will perform the infection studies with SARS-CoV-2 and its variants in both non-human immune engrafted (Specific Aim 1) and human immune engrafted mice (Specific Aim 2) and analyze the kinetics of virus clearance in real time using novel bioluminescent imaging technology. In addition, overall mouse health, viral load in tissues and inflammatory cytokines will be monitored. We will validate the optimal SARS-CoV-2 infection protocols with the NSG mouse stocks. Ultimately, these studies will determine the clearance and pathogenesis of SARS-CoV-2 and variants in the absence or presence of a human immune system.

项目概要 SARS-CoV-2 大流行对全球卫生界来说是前所未有的挑战，迫切需要了解 SARS-CoV-2 的发病机制和新出现的病毒变种的病理学。与 SARS-CoV-2 感染相关的疾病范围从无症状到严重，在某些情况下是致命的疾病与感染期间炎症细胞死亡、免疫系统激活和释放相关不幸的是，现有的 SARS-CoV-2 感染动物模型。由于基因组的多重差异，不能完全概括人类对病毒感染的免疫反应，我们补充提案的目标是分子和代谢途径以及免疫系统调节。对 PA-20-272 的反应是使用 Leonard 博士开发的一组转化人源化小鼠模型舒尔茨在杰克逊实验室研究 SARS-CoV-2 及其变异体的清除情况新兴的，并用于测试人类特异性疗法，以减轻与疾病相关的病理学此外，我们将使用一种新颖的体内生物发光成像方法来监测其动力学。 SARS-CoV-2 在体内的实时传播和清除我们组建了一个研究小组。 1) 创造新的人源化小鼠品系的丰富经验（杰克逊实验室的 Leonard Shultz 博士）实验室），2）将人类免疫系统移植到免疫缺陷小鼠体内（Dale Greiner 和 Michael 博士）麻省大学的 Brehm），3）人类病毒病原体的研究（耶鲁大学的 Priti Kumar 博士），以及 4）专业知识在体内实时成像病毒感染（耶鲁大学的 Pradeep Uchil 博士）。杰克逊实验室的资源有助于将有效模型快速分发给科学界社区，使我们的团队能够快速贡献 SARS-CoV-2 生物学知识。 Shultz 已经建立了表达人类 ACE2 的 NSG 小鼠新模型，并将把这些小鼠提供给 Dr. Shultz。 Kumar 博士和 Brehm 博士和 Greiner 博士将为小鼠植入人类 UCB CD34+。 HSC 并将向库马尔博士的实验室提供移植小鼠，库马尔博士和乌奇尔博士将进行这项工作。 SARS-CoV-2 及其变体在非人类免疫移植（具体目标 1）和人类免疫移植小鼠（具体目标 2）并使用实时分析病毒清除动力学此外，新型生物发光成像技术还可以检测小鼠的整体健康状况、组织中的病毒载量和我们将监测炎症细胞因子，以验证最佳的 SARS-CoV-2 感染方案。最终，这些研究将确定 SARS-CoV-2 的清除和发病机制。以及在不存在或存在人类免疫系统的情况下的变体。