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Status |
Public on May 26, 2020 |
Title |
Generation of human bronchial organoids for SARS-CoV-2 research |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
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 can 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 primary human bronchial epithelial cells (hBEpC) and examined whether they could be used as a model for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research. The hBO were found to contain basal, club, ciliated, and goblet cells. Also, 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 protein of SARS-CoV-2, were highly expressed. After hBO were infected with SARS-CoV-2, remarkable amplification of the viral genome and the expression of spike protein of the virus was confirmed. In addition, cytotoxicity and pyknosis cells were observed due to the virus infection. Furthermore, treatment with camostat, an inhibitor of TMPRSS2, reduced the viral copy number to 2% of the control group. RNA-seq analyses revealed genes whose expression was altered by SARS-CoV-2 infection and camostat treatment. These results suggest that our hBO are acceptable for SARS-CoV-2 infection and replication, but also can be used as a model for COVID-19 drug discovery.
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Overall design |
Total RNA was isolated from human bronchial organoids, primary human bronchial epithelial cells, and A549 cell, and then RNA-seq was performed. The human bronchial organoids were infected with SARS-CoV-2 in the presence of absence of camostat. At 5 days after the infection, total RNA was collected and RNA-seq was performed.
**Please note that the processed data *csv files have been updated on June 10, 2020**
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Contributor(s) |
Okamoto T, Takayama K |
Citation(s) |
35637255 |
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Submission date |
May 19, 2020 |
Last update date |
Jun 01, 2022 |
Contact name |
Daisuke Okuzaki |
E-mail(s) |
[email protected]
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Phone |
+81-6-6879-4935
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Organization name |
Osaka univ.
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Department |
Immunology Frontier Research Center
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Lab |
Human Immunology (Single Cell Genomics)
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Street address |
Yamadaoka 3-1
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City |
Suita |
State/province |
Osaka |
ZIP/Postal code |
565-0871 |
Country |
Japan |
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Platforms (2) |
GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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Samples (18)
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Relations |
BioProject |
PRJNA633783 |
SRA |
SRP262285 |
Supplementary file |
Size |
Download |
File type/resource |
GSE150819_RAW.tar |
2.4 Mb |
(http)(custom) |
TAR (of CSV) |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
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