Experimental validation of computerised models of clustering of platelet glycoprotein receptors that signal via tandem SH2 domain proteins

Maqsood, Zahra; Clark, Joanne C.; Martin, Eleyna M.; Cheung, Yam Fung Hilaire; Morán, Luis A.; Watson, Sean E. T.; Pike, Jeremy A.; Di, Ying; Poulter, Natalie S.; Slater, Alexandre; Lange, Bodo M. H.; Nieswandt, Bernhard; Eble, Johannes A.; Tomlinson, Mike G.; Owen, Dylan M.; Stegner, David; Bridge, Lloyd J.; Wierling, Christoph; Watson, Steve P.

Abstract

The clustering of platelet glycoprotein receptors with cytosolic YxxL and YxxM motifs, including GPVI, CLEC-2 and PEAR1, triggers activation via phosphorylation of the conserved tyrosine residues and recruitment of the tandem SH2 (Src homology 2) domain effector proteins, Syk and PI 3-kinase. We have modelled the clustering of these receptors with monovalent, divalent and tetravalent soluble ligands and with transmembrane ligands based on the law of mass action using ordinary differential equations and agent-based modelling. The models were experimentally evaluated in platelets and transfected cell lines using monovalent and multivalent ligands, including novel nanobody-based divalent and tetravalent ligands, by fluorescence correlation spectroscopy. Ligand valency, receptor number, receptor dimerisation, receptor phosphorylation and a cytosolic tandem SH2 domain protein act in synergy to drive receptor clustering. Threshold concentrations of a CLEC-2-blocking antibody and Syk inhibitor act in synergy to block platelet aggregation. This offers a strategy for countering the effect of avidity of multivalent ligands and in limiting off-target effects.

• Journal Article Type: Article
• Acceptance Date: Nov 4, 2022
• Online Publication Date: Nov 28, 2022
• Publication Date: Nov 28, 2022
• Deposit Date: Dec 9, 2022
• Publicly Available Date: Dec 12, 2022
• Journal: PLoS Computational Biology
• Print ISSN: 1553-734X
• Electronic ISSN: 1553-7358
• Publisher: Public Library of Science
• Peer Reviewed: Peer Reviewed
• Volume: 18
• Issue: 11
• Article Number: e1010708
• DOI: https://doi.org/10.1371/journal.pcbi.1010708
• Keywords: Computational Theory and Mathematics; Cellular and Molecular Neuroscience; Genetics; Molecular Biology; Ecology; Modeling and Simulation; Ecology, Evolution, Behavior and Systematics
• Public URL: https://uwe-repository.worktribe.com/output/10196043
• Publisher URL: https://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1010708