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

Autor: Salahudeen AA; 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., Choi SS; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Rustagi A; Division of Infectious Disease and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Zhu J; Stanford University School of Engineering, Department of Electrical Engineering, Stanford, CA, USA., van Unen V; 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., de la O SM; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Flynn RA; Stanford ChEM-H, Stanford University, Stanford, CA, USA.; Department of Chemistry, Stanford University, Stanford, CA, USA., Margalef-Català M; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA, USA., Santos AJM; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Ju J; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Batish A; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Usui T; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Zheng GXY; 10x Genomics, Pleasanton, CA, USA., Edwards CE; Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA., Wagar LE; 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., Luca V; Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA., Anchang B; Division of Biomedical Data Science, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Nagendran M; Division of Pulmonary, Allergy and Critical Care, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Nguyen K; Division of Gastroenterology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Hart DJ; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Terry JM; 10x Genomics, Pleasanton, CA, USA., Belgrader P; 10x Genomics, Pleasanton, CA, USA., Ziraldo SB; 10x Genomics, Pleasanton, CA, USA., Mikkelsen TS; 10x Genomics, Pleasanton, CA, USA., Harbury PB; Department of Biochemistry, Stanford University School of Medicine, Stanford, CA, USA., Glenn JS; Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.; Division of Gastroenterology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Garcia KC; Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA.; Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA, USA., Davis MM; 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.; Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA, USA., Baric RS; Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.; Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA., Sabatti C; Division of Biomedical Data Science, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA., Amieva MR; Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA, USA., Blish CA; Division of Infectious Disease and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA. cblish@stanford.edu.; Chan Zuckerberg Biohub, San Francisco, CA, USA. cblish@stanford.edu., Desai TJ; Division of Pulmonary, Allergy and Critical Care, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA. tdesai@stanford.edu., Kuo CJ; Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA. cjkuo@stanford.edu.
Jazyk: angličtina
Zdroj: Nature [Nature] 2020 Dec; Vol. 588 (7839), pp. 670-675. Date of Electronic Publication: 2020 Nov 25.
DOI: 10.1038/s41586-020-3014-1
Abstrakt: 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.
Databáze: MEDLINE
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