Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue.

Autor:	Aguiar JA; Dept of Biology, University of Waterloo, Waterloo, ON, Canada., Tremblay BJ; Dept of Biology, University of Waterloo, Waterloo, ON, Canada., Mansfield MJ; Genomics and Regulatory Systems Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Japan., Woody O; Faculty of Mathematics, University of Waterloo, Waterloo, ON, Canada., Lobb B; Dept of Biology, University of Waterloo, Waterloo, ON, Canada., Banerjee A; McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada., Chandiramohan A; Firestone Institute for Respiratory Health - Division of Respirology, Dept of Medicine, McMaster University, Hamilton, ON, Canada., Tiessen N; Firestone Institute for Respiratory Health - Division of Respirology, Dept of Medicine, McMaster University, Hamilton, ON, Canada., Cao Q; Firestone Institute for Respiratory Health - Division of Respirology, Dept of Medicine, McMaster University, Hamilton, ON, Canada., Dvorkin-Gheva A; McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada., Revill S; Firestone Institute for Respiratory Health - Division of Respirology, Dept of Medicine, McMaster University, Hamilton, ON, Canada., Miller MS; McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada.; Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON, Canada.; Dept of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada., Carlsten C; Division of Respiratory Medicine, Dept of Medicine, University of British Columbia, Vancouver, BC, Canada., Organ L; Nottingham NIHR Biomedical Research Centre, Respiratory Research Unit, University of Nottingham, Nottingham, UK., Joseph C; Nottingham NIHR Biomedical Research Centre, Respiratory Research Unit, University of Nottingham, Nottingham, UK., John A; Nottingham NIHR Biomedical Research Centre, Respiratory Research Unit, University of Nottingham, Nottingham, UK., Hanson P; Centre for Heart Lung Innovation, University of British Columbia, Vancouver, BC, Canada., Austin RC; Division of Nephrology, Dept of Medicine, McMaster University, Hamilton, ON, Canada., McManus BM; Centre for Heart Lung Innovation, University of British Columbia, Vancouver, BC, Canada., Jenkins G; Nottingham NIHR Biomedical Research Centre, Respiratory Research Unit, University of Nottingham, Nottingham, UK., Mossman K; McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada., Ask K; McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada.; Firestone Institute for Respiratory Health - Division of Respirology, Dept of Medicine, McMaster University, Hamilton, ON, Canada., Doxey AC; Dept of Biology, University of Waterloo, Waterloo, ON, Canada.; Firestone Institute for Respiratory Health - Division of Respirology, Dept of Medicine, McMaster University, Hamilton, ON, Canada.; A.C. Doxey and J.A. Hirota contributed equally to this article as lead authors and supervised the work., Hirota JA; Dept of Biology, University of Waterloo, Waterloo, ON, Canada hirotaja@mcmaster.ca.; McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada.; Firestone Institute for Respiratory Health - Division of Respirology, Dept of Medicine, McMaster University, Hamilton, ON, Canada.; Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON, Canada.; A.C. Doxey and J.A. Hirota contributed equally to this article as lead authors and supervised the work.
Jazyk:	angličtina
Zdroj:	The European respiratory journal [Eur Respir J] 2020 Sep 03; Vol. 56 (3). Date of Electronic Publication: 2020 Sep 03 (Print Publication: 2020).
DOI:	10.1183/13993003.01123-2020
Abstrakt:	In December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged, causing the coronavirus disease 2019 (COVID-19) pandemic. SARS-CoV, the agent responsible for the 2003 SARS outbreak, utilises angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) host molecules for viral entry. ACE2 and TMPRSS2 have recently been implicated in SARS-CoV-2 viral infection. Additional host molecules including ADAM17, cathepsin L, CD147 and GRP78 may also function as receptors for SARS-CoV-2.To determine the expression and in situ localisation of candidate SARS-CoV-2 receptors in the respiratory mucosa, we analysed gene expression datasets from airway epithelial cells of 515 healthy subjects, gene promoter activity analysis using the FANTOM5 dataset containing 120 distinct sample types, single cell RNA sequencing (scRNAseq) of 10 healthy subjects, proteomic datasets, immunoblots on multiple airway epithelial cell types, and immunohistochemistry on 98 human lung samples.We demonstrate absent to low ACE2 promoter activity in a variety of lung epithelial cell samples and low ACE2 gene expression in both microarray and scRNAseq datasets of epithelial cell populations. Consistent with gene expression, rare ACE2 protein expression was observed in the airway epithelium and alveoli of human lung, confirmed with proteomics. We present confirmatory evidence for the presence of TMPRSS2, CD147 and GRP78 protein in vitro in airway epithelial cells and confirm broad in situ protein expression of CD147 and GRP78 in the respiratory mucosa.Collectively, our data suggest the presence of a mechanism dynamically regulating ACE2 expression in human lung, perhaps in periods of SARS-CoV-2 infection, and also suggest that alternative receptors for SARS-CoV-2 exist to facilitate initial host cell infection. Competing Interests: Conflict of interest: J.A. Aguiar has nothing to disclose. Conflict of interest: B.J-M. Tremblay has nothing to disclose. Conflict of interest: M.J. Mansfield has nothing to disclose. Conflict of interest: O. Woody has nothing to disclose. Conflict of interest: B. Lobb has nothing to disclose. Conflict of interest: A. Banerjee has nothing to disclose. Conflict of interest: A. Chandiramohan has nothing to disclose. Conflict of interest: N. Tiessen has nothing to disclose. Conflict of interest: Q. Cao has nothing to disclose. Conflict of interest: A. Dvorkin-Gheva has nothing to disclose. Conflict of interest: S. Revill has nothing to disclose. Conflict of interest: M.S. Miller has nothing to disclose. Conflict of interest: C. Carlsten has nothing to disclose. Conflict of interest: L. Organ has nothing to disclose. Conflict of interest: C. Joseph has nothing to disclose. Conflict of interest: A. John has nothing to disclose. Conflict of interest: P. Hanson has nothing to disclose. Conflict of interest: R. Austin has a patent US7524826B2 issued to McMaster University and Hamilton Health Sciences Corp., McMaster University. Conflict of interest: B.M. McManus has nothing to disclose. Conflict of interest: G. Jenkins reports grants from AstraZeneca, Biogen, Galecto and GlaxoSmithKline; personal fees from Boehringer Ingelheim, Daewoong, Galapagos, Heptares, Promedior and Roche; grants and personal fees from Pliant; non-financial support from Redx and NuMedii; and is trustee for Action for Pulmonary Fibrosis, outside the submitted work. Conflict of interest: K. Mossman has nothing to disclose. Conflict of interest: K. Ask has nothing to disclose. Conflict of interest: A.C. Doxey has nothing to disclose. Conflict of interest: J.A. Hirota has nothing to disclose. (Copyright ©ERS 2020.)
Databáze:	MEDLINE
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