SARS-CoV-2 infection of primary human lung epithelium for COVID-19 modeling and drug discovery

Coronavirus disease 2019 (COVID-19) is the latest respiratory pandemic resulting from zoonotic transmission of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2). Severe symptoms include viral pneumonia secondary to infection and inflammation of the lower respiratory tract, in some cases causing death. We developed primary human lung epithelial infection models to understand responses of proximal and distal lung epithelium to SARS-CoV-2 infection. Differentiated air-liquid interface cultures of proximal airway epithelium and 3D organoid cultures of alveolar epithelium were readily infected by SARS-CoV-2 leading to an epithelial cell-autonomous proinflammatory response. We validated the efficacy of selected candidate COVID-19 drugs confirming that Remdesivir strongly suppressed viral infection/replication. We provide a relevant platform for studying COVID-19 pathobiology and for rapid drug screening against SARS-CoV-2 and future emergent respiratory pathogens. Overall design: We isolated primary lung alveolar AT2 cells and cultured in 3D, then infected with SARS-CoV2. RNA isolated from infected organoid for RNA-seq

2019冠状病毒病（Coronavirus disease 2019, COVID-19）是由严重急性呼吸综合征相关冠状病毒2（severe acute respiratory syndrome-related coronavirus 2, SARS-CoV-2）经动物源性传播引发的最新一次呼吸道大流行。其重症症状包括继发于下呼吸道感染与炎症的病毒性肺炎，部分病例可导致死亡。本研究构建原代人肺上皮感染模型，以探究近端与远端肺上皮细胞对SARS-CoV-2感染的应答反应。近端气道上皮的分化气液界面（air-liquid interface）培养物与肺泡上皮的3D类器官（organoid）培养物均可被SARS-CoV-2高效感染，进而引发上皮细胞自主性促炎应答。本研究验证了候选COVID-19治疗药物的疗效，证实瑞德西韦（Remdesivir）可强力抑制病毒感染与复制。本研究为COVID-19病理生物学研究，以及针对SARS-CoV-2与未来新发呼吸道病原体的快速药物筛选提供了适用平台。实验整体设计：我们分离原代肺肺泡AT2细胞并开展3D培养，随后以SARS-CoV-2感染细胞；从感染后的类器官中提取RNA用于RNA测序（RNA-seq）。