Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in context
Summary: Background: The COVID-19 pandemic is an infectious disease caused by SARS-CoV-2. The first step of SARS-CoV-2 infection is the recognition of angiotensin-converting enzyme 2 (ACE2) receptors by the receptor-binding domain (RBD) of the viral Spike (S) glycoprotein. Although the molecular an...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-01-01
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| Series: | EBioMedicine |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352396422005722 |
| _version_ | 1828084817494802432 |
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| author | Annarita Miluzio Alessandro Cuomo Chiara Cordiglieri Lorena Donnici Elisa Pesce Mauro Bombaci Matteo Conti Alessandra Fasciani Luigi Terracciano Lara Manganaro Mirco Toccafondi Alessandra Scagliola Stefania Oliveto Sara Ricciardi Renata Grifantini Raffaele De Francesco Sergio Abrignani Nicola Manfrini Stefano Biffo |
| author_facet | Annarita Miluzio Alessandro Cuomo Chiara Cordiglieri Lorena Donnici Elisa Pesce Mauro Bombaci Matteo Conti Alessandra Fasciani Luigi Terracciano Lara Manganaro Mirco Toccafondi Alessandra Scagliola Stefania Oliveto Sara Ricciardi Renata Grifantini Raffaele De Francesco Sergio Abrignani Nicola Manfrini Stefano Biffo |
| author_sort | Annarita Miluzio |
| collection | DOAJ |
| description | Summary: Background: The COVID-19 pandemic is an infectious disease caused by SARS-CoV-2. The first step of SARS-CoV-2 infection is the recognition of angiotensin-converting enzyme 2 (ACE2) receptors by the receptor-binding domain (RBD) of the viral Spike (S) glycoprotein. Although the molecular and structural bases of the SARS-CoV-2-RBD/hACE2 interaction have been thoroughly investigated in vitro, the relationship between hACE2 expression and in vivo infection is less understood. Methods: Here, we developed an efficient SARS-CoV-2-RBD binding assay suitable for super resolution microscopy and simultaneous hACE2 immunodetection and mapped the correlation between hACE2 receptor abundance and SARS-CoV-2-RBD binding, both in vitro and in human lung biopsies. Next, we explored the specific proteome of SARS-CoV-2-RBD/hACE2 through a comparative mass spectrometry approach. Findings: We found that only a minority of hACE2 positive spots are actually SARS-CoV-2-RBD binding sites, and that the relationship between SARS-CoV-2-RBD binding and hACE2 presence is variable, suggesting the existence of additional factors. Indeed, we found several interactors that are involved in receptor localization and viral entry and characterized one of them: SLC1A5, an amino acid transporter. High-resolution receptor-binding studies showed that co-expression of membrane-bound SLC1A5 with hACE2 predicted SARS-CoV-2 binding and entry better than hACE2 expression alone. SLC1A5 depletion reduces SARS-CoV-2 binding and entry. Notably, the Omicron variant is more efficient in binding hACE2 sites, but equally sensitive to SLC1A5 downregulation. Interpretation: We propose a method for mapping functional SARS-CoV-2 receptors in vivo. We confirm the existence of hACE2 co-factors that may contribute to differential sensitivity of cells to infection. Funding: This work was supported by an unrestricted grant from “Fondazione Romeo ed Enrica Invernizzi” to Stefano Biffo and by AIRC under MFAG 2021 - ID. 26178 project – P.I. Manfrini Nicola. |
| first_indexed | 2024-04-11T04:31:24Z |
| format | Article |
| id | doaj.art-607fc0df62fb49a49aba800f71f5b850 |
| institution | Directory Open Access Journal |
| issn | 2352-3964 |
| language | English |
| last_indexed | 2024-04-11T04:31:24Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | EBioMedicine |
| spelling | doaj.art-607fc0df62fb49a49aba800f71f5b8502022-12-29T04:13:19ZengElsevierEBioMedicine2352-39642023-01-0187104390Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in contextAnnarita Miluzio0Alessandro Cuomo1Chiara Cordiglieri2Lorena Donnici3Elisa Pesce4Mauro Bombaci5Matteo Conti6Alessandra Fasciani7Luigi Terracciano8Lara Manganaro9Mirco Toccafondi10Alessandra Scagliola11Stefania Oliveto12Sara Ricciardi13Renata Grifantini14Raffaele De Francesco15Sergio Abrignani16Nicola Manfrini17Stefano Biffo18National Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyDepartment of Experimental Oncology, IEO, European Institute of Oncology IRCCS, 20141, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyInstitute of Pathology, University Hospital Basel, 4031, Basel, SwitzerlandNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, Italy; Department of Biosciences, University of Milan, 20133, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, Italy; Department of Pharmacological and Biomolecular Sciences, University of Milan, 20133, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, Italy; Department of Clinical Sciences and Community Health, University of Milan, 20122, Milan, ItalyNational Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, Italy; Department of Biosciences, University of Milan, 20133, Milan, Italy; Corresponding author. National Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, Italy.National Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, Italy; Department of Biosciences, University of Milan, 20133, Milan, Italy; Corresponding author. National Institute of Molecular Genetics, ''Fondazione Romeo ed Enrica Invernizzi'', INGM, 20122, Milan, Italy.Summary: Background: The COVID-19 pandemic is an infectious disease caused by SARS-CoV-2. The first step of SARS-CoV-2 infection is the recognition of angiotensin-converting enzyme 2 (ACE2) receptors by the receptor-binding domain (RBD) of the viral Spike (S) glycoprotein. Although the molecular and structural bases of the SARS-CoV-2-RBD/hACE2 interaction have been thoroughly investigated in vitro, the relationship between hACE2 expression and in vivo infection is less understood. Methods: Here, we developed an efficient SARS-CoV-2-RBD binding assay suitable for super resolution microscopy and simultaneous hACE2 immunodetection and mapped the correlation between hACE2 receptor abundance and SARS-CoV-2-RBD binding, both in vitro and in human lung biopsies. Next, we explored the specific proteome of SARS-CoV-2-RBD/hACE2 through a comparative mass spectrometry approach. Findings: We found that only a minority of hACE2 positive spots are actually SARS-CoV-2-RBD binding sites, and that the relationship between SARS-CoV-2-RBD binding and hACE2 presence is variable, suggesting the existence of additional factors. Indeed, we found several interactors that are involved in receptor localization and viral entry and characterized one of them: SLC1A5, an amino acid transporter. High-resolution receptor-binding studies showed that co-expression of membrane-bound SLC1A5 with hACE2 predicted SARS-CoV-2 binding and entry better than hACE2 expression alone. SLC1A5 depletion reduces SARS-CoV-2 binding and entry. Notably, the Omicron variant is more efficient in binding hACE2 sites, but equally sensitive to SLC1A5 downregulation. Interpretation: We propose a method for mapping functional SARS-CoV-2 receptors in vivo. We confirm the existence of hACE2 co-factors that may contribute to differential sensitivity of cells to infection. Funding: This work was supported by an unrestricted grant from “Fondazione Romeo ed Enrica Invernizzi” to Stefano Biffo and by AIRC under MFAG 2021 - ID. 26178 project – P.I. Manfrini Nicola.http://www.sciencedirect.com/science/article/pii/S2352396422005722SpikeSuper resolution microscopyACE2 co-factorsRBDViral bindingViral entry |
| spellingShingle | Annarita Miluzio Alessandro Cuomo Chiara Cordiglieri Lorena Donnici Elisa Pesce Mauro Bombaci Matteo Conti Alessandra Fasciani Luigi Terracciano Lara Manganaro Mirco Toccafondi Alessandra Scagliola Stefania Oliveto Sara Ricciardi Renata Grifantini Raffaele De Francesco Sergio Abrignani Nicola Manfrini Stefano Biffo Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in context EBioMedicine Spike Super resolution microscopy ACE2 co-factors RBD Viral binding Viral entry |
| title | Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in context |
| title_full | Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in context |
| title_fullStr | Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in context |
| title_full_unstemmed | Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in context |
| title_short | Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2Research in context |
| title_sort | mapping of functional sars cov 2 receptors in human lungs establishes differences in variant binding and slc1a5 as a viral entry modulator of hace2research in context |
| topic | Spike Super resolution microscopy ACE2 co-factors RBD Viral binding Viral entry |
| url | http://www.sciencedirect.com/science/article/pii/S2352396422005722 |
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