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

Article

关键词: STEM-CELLS;    ALVEOLAR;    AIRWAY;    DIFFERENTIATION;    MOBILIZE;    BINDING;    MODEL;    WNT;   

DOI  :  10.1038/s41586-020-3014-1

来源: SCIE

【 摘 要 】

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.

【 授权许可】

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