Abstract
Coronavirus disease 2019 (COVID-19) is a disease that causes fatal disorders including severe pneumonia. To develop a therapeutic drug for COVID-19, a model that can reproduce the viral life cycle and evaluate the drug efficacy of anti-viral drugs is essential. In this study, we established a method to generate human bronchial organoids (hBO) from commercially available cryopreserved human bronchial epithelial cells and examined whether they could be used as a model for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research. Our hBO contain basal, club, ciliated, and goblet cells. Angiotensin-converting enzyme 2 (ACE2), which is a receptor for SARS-CoV-2, and transmembrane serine proteinase 2 (TMPRSS2), which is an essential serine protease for priming spike (S) protein of SARS-CoV-2, were highly expressed. After SARS-CoV-2 infection, not only the intracellular viral genome, but also progeny virus, cytotoxicity, pyknotic cells, and moderate increases of the type I interferon signal could be observed. Treatment with camostat, an inhibitor of TMPRSS2, reduced the viral copy number to 2% of the control group. Furthermore, the gene expression profile in SARS-CoV-2-infected hBO was obtained by performing RNA-seq analysis. In conclusion, we succeeded in generating hBO that can be used for SARS-CoV-2 research and COVID-19 drug discovery.
Competing Interest Statement
The authors have declared no competing interest.
Abbreviations
- 2D
- two-dimensional
- ACE2
- angiotensin-converting enzyme 2
- CC10
- club cell protein 10
- FGF
- fibroblast growth factor
- hBEpC
- human bronchial epithelial cells
- hBO
- human bronchial organoids
- IFN-I
- type I interferon
- IHC
- immunohistochemistry
- KRT5
- keratin 5
- LDH
- lactate dehydrogenase
- PSC
- pluripotent stem cell
- RdRp
- RNA-dependent RNA polymerase
- RNA
- seq RNA sequencing
- SARS-CoV-2
- severe acute respiratory syndrome coronavirus 2
- TMPRSS2
- transmembrane serine proteinase 2
- WHO
- World Health Organization
