Thromb Haemost 2020; 120(12): 1700-1715
DOI: 10.1055/s-0040-1721319
Blood Cells, Inflammation and Infection

Heparin Inhibits Cellular Invasion by SARS-CoV-2: Structural Dependence of the Interaction of the Spike S1 Receptor-Binding Domain with Heparin

Courtney J. Mycroft-West∗∗
1  Molecular and Structural Biosciences, School of Life Sciences, Keele University, Newcastle-Under-Lyme, Staffordshire, United Kingdom
,
Dunhao Su∗∗
2  Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom
,
Isabel Pagani∗∗
3  Viral Pathogenesis and Biosafety Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Milan, Italy
,
Timothy R. Rudd∗∗
4  Analytical and Biological Sciences Division, National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, United Kingdom
,
Stefano Elli∗∗
5  Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy
,
Neha S. Gandhi∗∗
6  School of Chemistry and Physics, Queensland University of Technology, Brisbane, Australia
7  Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
,
Scott E. Guimond∗∗
8  School of Medicine, Keele University, Newcastle-Under-Lyme, Staffordshire, United Kingdom
,
Gavin J. Miller
9  School of Chemical and Physical Sciences, Keele University, Newcastle-Under-Lyme, Staffordshire, United Kingdom
,
Maria C. Z. Meneghetti
10  Biochemistry Department, Federal University of São Paulo (UNIFESP), São Paulo, SP Brazil
,
Helena B. Nader
10  Biochemistry Department, Federal University of São Paulo (UNIFESP), São Paulo, SP Brazil
,
Yong Li
2  Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom
,
Quentin M. Nunes
11  Department of Molecular and Clinical Cancer Medicine, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom
,
Patricia Procter
1  Molecular and Structural Biosciences, School of Life Sciences, Keele University, Newcastle-Under-Lyme, Staffordshire, United Kingdom
,
Nicasio Mancini
12  Università Vita-Salute San Raffaele, Milan, Italy
,
Massimo Clementi
12  Università Vita-Salute San Raffaele, Milan, Italy
,
Antonella Bisio
5  Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy
,
Nicholas R. Forsyth
13  Guy Hilton Research Centre, School of Pharmacy and Bioengineering, Keele University, Hartshill, Stoke-on-Trent, Staffordshire, United Kingdom
,
Vito Ferro
14  School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, Australia
15  Australian Infectious Diseases Research Centre, The University of Queensland, Brisbane, Australia
,
Jeremy E. Turnbull
2  Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom
,
Marco Guerrini
5  Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy
,
David G. Fernig
2  Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom
,
Elisa Vicenzi
3  Viral Pathogenesis and Biosafety Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Milan, Italy
,
Edwin A. Yates
1  Molecular and Structural Biosciences, School of Life Sciences, Keele University, Newcastle-Under-Lyme, Staffordshire, United Kingdom
2  Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom
,
Marcelo A. Lima
1  Molecular and Structural Biosciences, School of Life Sciences, Keele University, Newcastle-Under-Lyme, Staffordshire, United Kingdom
,
1  Molecular and Structural Biosciences, School of Life Sciences, Keele University, Newcastle-Under-Lyme, Staffordshire, United Kingdom
2  Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom
› Author Affiliations
Funding This study was funded by the Biotechnology and Biological Sciences Research Council, UK (BB/L023717/1; BIV-HVB-2020/07/SKIDMORE; BB/S009787/1) and the Fundação de Amparo à Pesquisa do Estado de São Paulo, Brazil (2020/04899-1).

Abstract

The dependence of development and homeostasis in animals on the interaction of hundreds of extracellular regulatory proteins with the peri- and extracellular glycosaminoglycan heparan sulfate (HS) is exploited by many microbial pathogens as a means of adherence and invasion. Heparin, a widely used anticoagulant drug, is structurally similar to HS and is a common experimental proxy. Exogenous heparin prevents infection by a range of viruses, including S-associated coronavirus isolate HSR1. Here, we show that heparin inhibits severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) invasion of Vero cells by up to 80% at doses achievable through prophylaxis and, particularly relevant, within the range deliverable by nebulisation. Surface plasmon resonance and circular dichroism spectroscopy demonstrate that heparin and enoxaparin, a low-molecular-weight heparin which is a clinical anticoagulant, bind and induce a conformational change in the spike (S1) protein receptor-binding domain (S1 RBD) of SARS-CoV-2. A library of heparin derivatives and size-defined fragments were used to probe the structural basis of this interaction. Binding to the RBD is more strongly dependent on the presence of 2-O or 6-O sulfate groups than on N-sulfation and a hexasaccharide is the minimum size required for secondary structural changes to be induced in the RBD. It is likely that inhibition of viral infection arises from an overlap between the binding sites of heparin/HS on S1 RBD and that of the angiotensin-converting enzyme 2. The results suggest a route for the rapid development of a first-line therapeutic by repurposing heparin and its derivatives as antiviral agents against SARS-CoV-2 and other members of the Coronaviridae.

These authors are joint first authors.


∗∗ Senior authors.


Supplementary Material



Publication History

Received: 06 June 2020

Accepted: 13 October 2020

Publication Date:
23 December 2020 (online)

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