TY - JOUR
KW - Adult Stem Cells
KW - Influenza virus
KW - SARS-CoV-2
AU - Ameen A. Salahudeen
AU - Shannon S. Choi
AU - Arjun Rustagi
AU - Junjie Zhu
AU - Vincent van Unen
AU - Sean M. de la O
AU - Ryan A. Flynn
AU - Mar Margalef-Català
AU - António J. M. Santos
AU - Jihang Ju
AU - Arpit Batish
AU - Tatsuya Usui
AU - Grace X. Y. Zheng
AU - Caitlin E. Edwards
AU - Lisa E. Wagar
AU - Vincent Luca
AU - Benedict Anchang
AU - Monica Nagendran
AU - Khanh Nguyen
AU - Daniel J. Hart
AU - Jessica M. Terry
AU - Phillip Belgrader
AU - Solongo B. Ziraldo
AU - Tarjei S. Mikkelsen
AU - Pehr B. Harbury
AU - Jeffrey S. Glenn
AU - K. Christopher Garcia
AU - Mark M. Davis
AU - Ralph S. Baric
AU - Chiara Sabatti
AU - Manuel R. Amieva
AU - Catherine A. Blish
AU - Tushar J. Desai
AU - Calvin J. Kuo
AB - 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 TNFRSF12Ahi 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.
BT - Nature
DA - 2020-12
DO - 10.1038/s41586-020-3014-1
IS - 7839
LA - en
N2 - 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 TNFRSF12Ahi 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.
PY - 2020
SP - 670
EP - 675
T2 - Nature
TI - Progenitor identification and SARS-CoV-2 infection in human distal lung organoids
UR - https://www.nature.com/articles/s41586-020-3014-1
VL - 588
Y2 - 2023-08-09
SN - 1476-4687
ER -