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

Suzuki, T.; Ito, Y.; Sakai, Y.; Saito, A.; Okuzaki, D.; Motooka, D.; Minami, S.; Kobayashi, T.; Yamamoto, T.; Okamoto, T.; Takayama, K.

doi:10.1101/2020.05.25.115600

bioRxiv · 10.1101/2020.05.25.115600

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

Suzuki, T.·Ito, Y.·Sakai, Y.·Saito, A.·Okuzaki, D.·Motooka, D.·Minami, S.·Kobayashi, T.·Yamamoto, T.·Okamoto, T.·Takayama, K.

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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. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/115600v2_ufig1.gif" ALT="Figure 1"> View larger version (99K): org.highwire.dtl.DTLVardef@13a6908org.highwire.dtl.DTLVardef@1c59300org.highwire.dtl.DTLVardef@362167org.highwire.dtl.DTLVardef@1cb31ed_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Suzuki, T., Ito, Y., Sakai, Y., Saito, A., Okuzaki, D., Motooka, D., Minami, S., Kobayashi, T., Yamamoto, T., Okamoto, T., Takayama, K.. 2020-05-26. Generation of human bronchial organoids for SARS-CoV-2 research. https://doi.org/10.1101/2020.05.25.115600

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