Generation of human bronchial organoids for SARS-CoV-2 research

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...

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Bibliographic Details
Published in:bioRxiv 2020-06
Main Authors: Suzuki, Tatsuya, Itoh, Yumi, Sakai, Yusuke, Saito, Akatsuki, Okuzaki, Daisuke, Motooka, Daisuke, Minami, Shohei, Kobayashi, Takeshi, Yamamoto, Takuya, Okamoto, Toru, Takayama, Kazuo
Format: Article
Language:English
Subjects:
ACE2
Angiotensin
Angiotensin-converting enzyme 2
Antiviral agents
Copy number
Coronaviruses
COVID-19
Cryopreservation
Cytotoxicity
Epithelial cells
Gene expression
Genomes
Goblet cells
Interferon
Life cycles
Organoids
Peptidyl-dipeptidase A
Proteinase
Ribonucleic acid
RNA
Serine
Serine proteinase
Severe acute respiratory syndrome coronavirus 2
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Summary: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. Figure1 Figure1 * Download figure * Open in new tab Competing Interest Statement The authors have declared no competing interest. Footnotes * https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE150819 * 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
DOI:10.1101/2020.05.25.115600