Tags

Type your tag names separated by a space and hit enter

In well-differentiated primary human bronchial epithelial cells, TGF-β1 and TGF-β2 induce expression of furin.
Am J Physiol Lung Cell Mol Physiol. 2021 02 01; 320(2):L246-L253.AJ

Abstract

The COVID-19 pandemic is an ongoing threat to public health. Since the identification of COVID-19, the disease caused by SARS-CoV-2, no drugs have been developed to specifically target SARS-CoV-2. To develop effective and safe treatment options, a better understanding of cellular mechanisms underlying SARS-CoV-2 infection is required. To fill this knowledge gap, researchers require reliable experimental systems that express the host factor proteins necessary for the cellular entry of SARS-CoV-2. These proteins include the viral receptor, angiotensin-converting enzyme 2 (ACE2), and the proteases, transmembrane serine protease 2 (TMPRSS2) and furin. A number of studies have reported cell-type-specific expression of the genes encoding these molecules. However, less is known about the protein expression of these molecules. We assessed the suitability of primary human bronchial epithelial (HBE) cells maintained in an air-liquid interface (ALI) as an experimental system for studying SARS-CoV-2 infection in vitro. During cellular differentiation, we measured the expression of ACE2, TMPRSS2, and furin over progressive ALI days by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot, and immunofluorescence staining. We also explored the effect of the fibrotic cytokine TGF-β on the expression of these proteins in well-differentiated HBE cells. Like ACE2, TMPRSS2 and furin proteins are localized in differentiated ciliated cells, as confirmed by immunofluorescence staining. These data suggest that well-differentiated HBE cells maintained in ALI are a reliable in vitro system for investigating cellular mechanisms of SARS-CoV-2 infection. We further identified that the profibrotic mediators, TGF-β1 and TGF-β2, increase the expression of furin, which is a protease required for the cellular entry of SARS-CoV-2.

Links

PMC Free PDF

Authors+Show Affiliations

O'Sullivan MJ
Program in Molecular and Integrative Physiological Sciences, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts.
Mitchel JA
Program in Molecular and Integrative Physiological Sciences, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts.
Mwase C
Program in Molecular and Integrative Physiological Sciences, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts.
McGill M
Program in Molecular and Integrative Physiological Sciences, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts.
Kanki P
Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts.
Park JA
Program in Molecular and Integrative Physiological Sciences, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts.

MeSH

Angiotensin-Converting Enzyme 2BronchiCOVID-19Cell DifferentiationCells, CulturedDisease SusceptibilityEpithelial CellsFurinGene ExpressionHost Microbial InteractionsHumansModels, BiologicalPandemicsRNA, MessengerSARS-CoV-2Serine EndopeptidasesTransforming Growth Factor beta1Transforming Growth Factor beta2Virus Internalization

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33174447

Citation

O'Sullivan, Michael J., et al. "In Well-differentiated Primary Human Bronchial Epithelial Cells, TGF-β1 and TGF-β2 Induce Expression of Furin." American Journal of Physiology. Lung Cellular and Molecular Physiology, vol. 320, no. 2, 2021, pp. L246-L253.
O'Sullivan MJ, Mitchel JA, Mwase C, et al. In well-differentiated primary human bronchial epithelial cells, TGF-β1 and TGF-β2 induce expression of furin. Am J Physiol Lung Cell Mol Physiol. 2021;320(2):L246-L253.
O'Sullivan, M. J., Mitchel, J. A., Mwase, C., McGill, M., Kanki, P., & Park, J. A. (2021). In well-differentiated primary human bronchial epithelial cells, TGF-β1 and TGF-β2 induce expression of furin. American Journal of Physiology. Lung Cellular and Molecular Physiology, 320(2), L246-L253. https://doi.org/10.1152/ajplung.00423.2020
O'Sullivan MJ, et al. In Well-differentiated Primary Human Bronchial Epithelial Cells, TGF-β1 and TGF-β2 Induce Expression of Furin. Am J Physiol Lung Cell Mol Physiol. 2021 02 1;320(2):L246-L253. PubMed PMID: 33174447.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - In well-differentiated primary human bronchial epithelial cells, TGF-β1 and TGF-β2 induce expression of furin. AU - O'Sullivan,Michael J, AU - Mitchel,Jennifer A, AU - Mwase,Chimwemwe, AU - McGill,Maureen, AU - Kanki,Phyllis, AU - Park,Jin-Ah, Y1 - 2020/11/11/ PY - 2020/11/12/pubmed PY - 2021/2/26/medline PY - 2020/11/11/entrez KW - ACE2 KW - COVID-19 KW - SARS-CoV-2 KW - TGF-β KW - TMPRSS2 KW - furin SP - L246 EP - L253 JF - American journal of physiology. Lung cellular and molecular physiology JO - Am J Physiol Lung Cell Mol Physiol VL - 320 IS - 2 N2 - The COVID-19 pandemic is an ongoing threat to public health. Since the identification of COVID-19, the disease caused by SARS-CoV-2, no drugs have been developed to specifically target SARS-CoV-2. To develop effective and safe treatment options, a better understanding of cellular mechanisms underlying SARS-CoV-2 infection is required. To fill this knowledge gap, researchers require reliable experimental systems that express the host factor proteins necessary for the cellular entry of SARS-CoV-2. These proteins include the viral receptor, angiotensin-converting enzyme 2 (ACE2), and the proteases, transmembrane serine protease 2 (TMPRSS2) and furin. A number of studies have reported cell-type-specific expression of the genes encoding these molecules. However, less is known about the protein expression of these molecules. We assessed the suitability of primary human bronchial epithelial (HBE) cells maintained in an air-liquid interface (ALI) as an experimental system for studying SARS-CoV-2 infection in vitro. During cellular differentiation, we measured the expression of ACE2, TMPRSS2, and furin over progressive ALI days by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot, and immunofluorescence staining. We also explored the effect of the fibrotic cytokine TGF-β on the expression of these proteins in well-differentiated HBE cells. Like ACE2, TMPRSS2 and furin proteins are localized in differentiated ciliated cells, as confirmed by immunofluorescence staining. These data suggest that well-differentiated HBE cells maintained in ALI are a reliable in vitro system for investigating cellular mechanisms of SARS-CoV-2 infection. We further identified that the profibrotic mediators, TGF-β1 and TGF-β2, increase the expression of furin, which is a protease required for the cellular entry of SARS-CoV-2. SN - 1522-1504 UR - https://www.unboundmedicine.com/medline/citation/33174447/In_well_differentiated_primary_human_bronchial_epithelial_cells_TGF_β1_and_TGF_β2_induce_expression_of_furin_ DB - PRIME DP - Unbound Medicine ER -