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
View: Text | PDF
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

×

Figure 1

CRISPRa screen against authentic SARS-CoV-2.

Options: View larger image (or click on image) Download as PowerPoint
CRISPRa screen against authentic SARS-CoV-2. (A) Schematic showing whole...
(A) Schematic showing whole-genome CRISPRa screen in HEK293-ACE2 cells using the Weissman library (https://weissman.wi.mit.edu/crispr/). Cells were transduced with lentiviral pools encoding individual activation sgRNAs tiling the genome. Cells were then inoculated with authentic SARS-CoV-2 or parallel controls, and guides promoting SARS-CoV-2 were identified by sequencing. (N = 2.) (B) Top genes by local FDR (locFDR) (<0.4 considered significant) identified after selection. See also Supplemental Table 1. (C) Plot showing sgRNA enrichment (log fold change) versus gene ranking. (D and E) Top pathways identified using Ingenuity Pathway Analysis (IPA) (D) or KEGG pathways (E). (F and G) Independent validation of top hits including cytosolic (ARL4C, LRRC29, RPS16) or mitochondrial proteins (MRPS7) (F), and membrane proteins promoting SARS-CoV-2 resistance (48 hours after inoculation) (G). Significance was determined by 2-way ANOVA and Dunnett’s test. (N = 3.) (H) Diagram of SARS-CoV-2 FACS spike binding assay. (I) CRISPRa-driven expression of SELP/P selectin promotes binding to SARS-CoV-2 spike protein. Significance was obtained by 1-way ANOVA and Šidák’s test, *P < 0.05. (N = 3.)