Organoid Core B

摘要 原代人类类器官模型是越来越多地用于体外感染性疾病建模的平台。 由新型冠状病毒SARS-CoV-2引起的COVID-19大流行，对 因此，迫切需要公共卫生和生理体外感染模型。这一补充请求 对于U19 AI 116484，斯坦福大学肠道疾病新型替代模型系统合作中心 （斯坦福大学NAMSED），要求资助使用新的人类肺来创建SARS-CoV-2感染的新模型 类器官技术，与公认的冠状病毒权威机构--哈佛大学的Ralph Baric博士合作。这些 研究利用无饲养层、化学定义的人肺类器官利用SARS-CoV-2感染类器官 系统（卡尔文郭实验室），具有整合免疫组分的肺类器官（卡尔文郭），稳健的方法 肺类器官极性的顶部-底部反转（Manuel Amieva），BSL 3单细胞RNA-seq（Catherine Blish） 和SARS-CoV-2-GFP指示菌株和BSL 3设施（Ralph Baric）。SARS-CoV-2肺部感染 类器官将出现在斯坦福大学和斯坦福大学的BSL 3容器中，以比较顶端与基底感染 途径，记录上皮感染如何启动二次免疫应答，并总体产生改善的 SARS-CoV-2-GFP感染的3D生理模型与治疗筛选相关。

ABSTRACT Primary human organoid models are an increasingly deployed platform for in vitro infectious disease modeling. The COVID-19 pandemic, engendered by the novel coronavirus SARS-CoV-2, represents a grave threat to public health and physiologic in vitro infection models are therefore urgently needed. This supplement request for U19AI116484, Stanford Cooperative Center for Novel, Alternative Model Systems for Enteric Diseases (Stanford NAMSED), requests funding to create new models for SARS-CoV-2 infection using novel human lung organoid technologies in collaboration with Dr. Ralph Baric at UNC, a recognized coronavirus authority. These studies exploit SARS-CoV-2 infection of organoids using a feeder-free, chemically defined human lung organoid system (Calvin Kuo lab), lung organoids with integrated immune components (Calvin Kuo), methods for robust apical-basal inversion of lung organoid polarity (Manuel Amieva), BSL3 single cell RNA-seq (Catherine Blish) and SARS-CoV-2-GFP indicator strains and BSL3 facilities (Ralph Baric). The SARS-CoV-2 infection of lung organoids will occur in BSL3 containment at both UNC and Stanford to compare apical versus basal infection routes, document how epithelial infection initiates secondary immune responses, and overall generate improved 3D physiological models of SARS-CoV-2-GFP infection relevant to therapeutics screening.