P selectin promotes SARS-CoV-2 interactions with platelets and the endothelium
Cesar L. Moreno, Fernanda V.S. Castanheira, Alberto Ospina Stella, Felicity Chung, Anupriya Aggarwal, Alexander J. Cole, Lipin Loo, Alexander Dupuy, Yvonne X. Kong, Lejla Hagimola, Jemma Fenwick, Paul R. Coleman, Rebecca Carr, Tian Y. Du, Tim Ison, Michelle Newton, Maxwell P. Bui-Marinos, Scott B. Cohen, Jennifer A. Corcoran, Daniel Hesselson, Jennifer R. Gamble, Freda H. Passam, Stuart G. Turville, Paul Kubes, G. Gregory Neely
Cesar L. Moreno, Fernanda V.S. Castanheira, Alberto Ospina Stella, Felicity Chung, Anupriya Aggarwal, Alexander J. Cole, Lipin Loo, Alexander Dupuy, Yvonne X. Kong, Lejla Hagimola, Jemma Fenwick, Paul R. Coleman, Rebecca Carr, Tian Y. Du, Tim Ison, Michelle Newton, Maxwell P. Bui-Marinos, Scott B. Cohen, Jennifer A. Corcoran, Daniel Hesselson, Jennifer R. Gamble, Freda H. Passam, Stuart G. Turville, Paul Kubes, G. Gregory Neely
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Research Article Infectious disease Virology

P selectin promotes SARS-CoV-2 interactions with platelets and the endothelium

Abstract

The physiology of SARS-CoV-2 virus/host interactions is not well understood. To better understand host/virus interactions, we performed a CRISPR activation screen to identify host genes that confer resistance to authentic SARS-CoV-2. This highlighted 34 new candidate genes that may alter the course of infection. We validated that 7 of these genes can suppress authentic SARS-CoV-2 infection, including the innate immune receptor P selectin, which increases SARS-CoV-2 spike-dependent binding to cells, while protecting from infection. P selectin also promotes binding to SARS-CoV-2 variants, SARS-CoV-1, and Middle East respiratory syndrome spike proteins, suggesting a general role for P selectin in highly pathogenic coronavirus infections. Importantly, P selectin protein expression driven by synthetic mRNA can block SARS-CoV-2 infection. Naturally, P selectin is expressed on platelets, and we show that it promotes spike-mediated platelet aggregation. P selectin is also expressed on the endothelium, where SARS-CoV-2 spike interactions are also P selectin dependent. In vivo, SARS-CoV-2 uses P selectin to home to capillary beds where the virus interacts with platelets and endothelium, and blocking this interaction can clear vascular-associated pulmonary SARS-CoV-2.

Authors

Cesar L. Moreno, Fernanda V.S. Castanheira, Alberto Ospina Stella, Felicity Chung, Anupriya Aggarwal, Alexander J. Cole, Lipin Loo, Alexander Dupuy, Yvonne X. Kong, Lejla Hagimola, Jemma Fenwick, Paul R. Coleman, Rebecca Carr, Tian Y. Du, Tim Ison, Michelle Newton, Maxwell P. Bui-Marinos, Scott B. Cohen, Jennifer A. Corcoran, Daniel Hesselson, Jennifer R. Gamble, Freda H. Passam, Stuart G. Turville, Paul Kubes, G. Gregory Neely

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Figure 5

Endothelial cells use P selectin to bind SARS-CoV-2 spike.

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Endothelial cells use P selectin to bind SARS-CoV-2 spike. (A) Schematic...
(A) Schematic of microfluidic devices used to study SARS-CoV-2 pseudovirus binding to P selectin under shear conditions. (B) Representative images of capillary lined with cells expressing GFP or P selectin–GFP after 1 hour of flow with SARS-CoV-2 pseudovirus–containing medium. Scale bars: 100 μm. (C) Representative images detailing accumulation of labeled pseudovirus particles in GFP- or P selectin–GFP–expressing cells. Scale bars: 10 μm. (D) Automated quantification of total DiD area/cells within the capillary. Significance was determined by 1-way ANOVA and Dunnett’s test, *P < 0.05. (N = 3.) (E) CRISPR targeting of SELP in primary endothelial cells (HUVECs) reduces P selectin staining (scale bar: 200 μm), quantified by flow cytometry in F. Significance was determined by 1-way ANOVA and Dunnett’s test, *P < 0.05. (N = 4.) (G) CRISPR-targeted SELP-edited HUVECs bind less SARS-CoV-2 spike protein. Significance was determined by 1-way ANOVA and Dunnett’s test, *P < 0.05, **P < 0.01. (N = 4.)