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 2

P selectin interaction with pathogenic coronavirus spike proteins.

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P selectin interaction with pathogenic coronavirus spike proteins. (A) E...
(A) Ectopic ACE2 or P selectin expression strategy. Scale bars: 60 µm. (B) Representative flow cytometry plots showing binding of P selectin to SARS-CoV-2 spike protein. (C) Quantification of B. Significance was determined by paired 2-tailed t test, **P < 0.01. (N = 3.) (D) Schematic of DiD-labeled pseudolentivirus assay used in E and F. (E and F) Flow cytometry histograms (E) and quantification of binding intensity (F) presented for pseudovirus expressing MERS, SARS-CoV-1, and SARS-CoV-2 A2.2 and Delta variants. MFI, mean fluorescence intensity. Significance was determined by 1-way ANOVA and Dunnett’s test, *P < 0.05, **P < 0.01. (N = 3.) (G) Binding sites (red) for blocking antibodies AZD1061 (cilgavimab), AZD8895 (tixagevimab), and S309 (sotrovimab) and predicted binding site for P selectin (blue) on SARS-CoV-2 spike glycoprotein (Protein Data Bank: 7WK2) (64). (H) Key predicted interactive regions between P selectin and spike RBD. C-type lectin-like domain, CTLD. (I) Binding of antibody-blocked spike protein in HEK293T cells expressing P selectin. Significance was determined by 1-way ANOVA and Dunnett’s test, **P < 0.01, ***P < 0.001. (N = 3).