Progenitor identification and SARS-CoV-2 infection in human distal lung organoids,Nature

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Progenitor identification and SARS-CoV-2 infection in human distal lung organoids
Nature ( IF 50.5 ) Pub Date : 2020-11-25 , DOI: 10.1038/s41586-020-3014-1
Ameen A Salahudeen _{1,

2} , Shannon S Choi ₁ , Arjun Rustagi ₃ , Junjie Zhu ₄ , Vincent van Unen _{5,

6} , Sean M de la O ₁ , Ryan A Flynn _{7,

8} , Mar Margalef-Català ₉ , António J M Santos ₁ , Jihang Ju ₁ , Arpit Batish ₁ , Tatsuya Usui ₁ , Grace X Y Zheng ₁₀ , Caitlin E Edwards ₁₁ , Lisa E Wagar _{5,

6} , Vincent Luca ₁₂ , Benedict Anchang ₁₃ , Monica Nagendran ₁₄ , Khanh Nguyen ₁₅ , Daniel J Hart ₁ , Jessica M Terry ₁₀ , Phillip Belgrader ₁₀ , Solongo B Ziraldo ₁₀ , Tarjei S Mikkelsen ₁₀ , Pehr B Harbury ₁₆ , Jeffrey S Glenn _{5,

15} , K Christopher Garcia _{12,

17} , Mark M Davis _{5,

6,

17} , Ralph S Baric _{11,

18} , Chiara Sabatti ₁₃ , Manuel R Amieva _{5,

9} , Catherine A Blish _{3,

19} , Tushar J Desai ₁₄ , Calvin J Kuo ₁

Affiliation

Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Division of Hematology and Oncology, Department of Medicine, University of Illinois at Chicago College of Medicine, Chicago, IL, USA.
Division of Infectious Disease and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Stanford University School of Engineering, Department of Electrical Engineering, Stanford, CA, USA.
Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.
Stanford Institute of Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, CA, USA.
Stanford ChEM-H, Stanford University, Stanford, CA, USA.
Department of Chemistry, Stanford University, Stanford, CA, USA.
Department of Pediatrics, Stanford University School of Medicine, Stanford, CA, USA.
10x Genomics, Pleasanton, CA, USA.
Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA.
Division of Biomedical Data Science, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Division of Pulmonary, Allergy and Critical Care, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Division of Gastroenterology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Department of Biochemistry, Stanford University School of Medicine, Stanford, CA, USA.
Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA, USA.
Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Chan Zuckerberg Biohub, San Francisco, CA, USA.

The distal lung contains terminal bronchioles and alveoli that facilitate gas exchange. Three-dimensional in vitro human distal lung culture systems would strongly facilitate the investigation of pathologies such as interstitial lung disease, cancer and coronavirus disease 2019 (COVID-19) pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here we describe the development of a long-term feeder-free, chemically defined culture system for distal lung progenitors as organoids derived from single adult human alveolar epithelial type II (AT2) or KRT5 + basal cells. AT2 organoids were able to differentiate into AT1 cells, and basal cell organoids developed lumens lined with differentiated club and ciliated cells. Single-cell analysis of KRT5 + cells in basal organoids revealed a distinct population of ITGA6 + ITGB4 + mitotic cells, whose offspring further segregated into a TNFRSF12A hi subfraction that comprised about ten per cent of KRT5 + basal cells. This subpopulation formed clusters within terminal bronchioles and exhibited enriched clonogenic organoid growth activity. We created distal lung organoids with apical-out polarity to present ACE2 on the exposed external surface, facilitating infection of AT2 and basal cultures with SARS-CoV-2 and identifying club cells as a target population. This long-term, feeder-free culture of human distal lung organoids, coupled with single-cell analysis, identifies functional heterogeneity among basal cells and establishes a facile in vitro organoid model of human distal lung infections, including COVID-19-associated pneumonia. A long-term culture method for organoids derived from single adult human lung cells is used to identify progenitor cells and study SARS-CoV-2 infection.

中文翻译：

人类远端肺类器官中的祖细胞鉴定和 SARS-CoV-2 感染

远端肺含有终末细支气管和肺泡，促进气体交换。三维体外人体远端肺培养系统将极大地促进间质性肺病、癌症和由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）肺炎等病理学的研究。在这里，我们描述了一种长期无饲养层、化学成分明确的培养系统的开发，用于远端肺祖细胞，作为源自单个成人 II 型肺泡上皮 (AT2) 或 KRT5 + 基底细胞的类器官。 AT2 类器官能够分化为 AT1 细胞，基底细胞类器官发育出内衬分化的棒状细胞和纤毛细胞的管腔。对基底类器官中 KRT5 + 细胞的单细胞分析揭示了 ITGA6 + ITGB4 + 有丝分裂细胞的独特群体，其后代进一步分离为 TNFRSF12A hi 亚组分，约占 KRT5 + 基底细胞的 10%。该亚群在终末细支气管内形成簇，并表现出丰富的克隆形成类器官生长活性。我们创建了具有顶端向外极性的远端肺类器官，以在暴露的外表面上呈现 ACE2，促进 SARS-CoV-2 感染 AT2 和基础培养物，并将俱乐部细胞识别为目标群体。这种人类远端肺类器官的长期、无饲养层培养，结合单细胞分析，识别了基底细胞之间的功能异质性，并建立了人类远端肺部感染（包括 COVID-19 相关肺炎）的简便体外类器官模型。使用源自单个成人肺细胞的类器官的长期培养方法来鉴定祖细胞并研究 SARS-CoV-2 感染。

更新日期：2020-11-25

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