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

项目总结 SARS-CoV-2大流行对全球卫生界来说是一个前所未有的挑战， 迫切需要了解SARS-CoV-2和新出现的病毒变异的发病机制。《病理学》 与SARS-CoV-2感染有关的疾病从无症状到严重，在某些情况下是致命的疾病， 与感染过程中炎性细胞死亡、免疫系统激活和释放 炎性细胞因子或“细胞因子风暴”。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， 分子和代谢途径，以及免疫系统调节。我们的补充提案的目标是 对PA-20-272反应是使用伦纳德博士开发的一组翻译的人源化小鼠模型 舒尔茨在杰克逊实验室研究SARS-CoV-2和持续不断的变异株的清除 正在出现，并用于测试人类特有的治疗方法，以减轻与 新冠肺炎。此外，我们将使用一种新的在体生物发光成像方法来监测 SARS-CoV-2在体内实时传播和清除。我们已经组建了一支调查小组， 在创造新的人性化小鼠品系方面有丰富的经验(杰克逊大学的伦纳德·舒尔茨博士 实验室)，2)将人类免疫系统移植到免疫缺陷小鼠(Dale Greiner和Michael博士 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 在活体内对病毒感染进行实时成像(耶鲁大学普拉迪普·乌奇尔博士)。我们将利用 杰克逊实验室的资源，以促进将有效的模型迅速分发给科学工作者 社区，使我们的团队能够迅速为SARS-CoV-2生物学知识做出贡献。Dr。 舒尔茨已经建立了表达人ACE2的NSG小鼠的新模型，并将把这些小鼠提供给Dr。 库马尔和布雷姆博士和格雷纳。布雷姆和格雷纳博士将把人脐血CD34+植入小鼠体内 并将向库马尔博士的实验室提供移植的小鼠。Kumar博士和Uchil博士将表演 非人免疫移植中SARS-CoV-2及其变异体的感染研究(特定目标1)和 人免疫移植小鼠(特异性靶点2)，并实时分析病毒清除动力学 新型生物发光成像技术。此外，小鼠的总体健康、组织中的病毒载量和 将监测炎性细胞因子。我们将验证最佳SARS-CoV-2感染方案 NSG老鼠库存。最终，这些研究将确定SARS-CoV-2的清除和发病机制。 以及在没有或存在人类免疫系统的情况下的变体。

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.