Platelet Adhesion to Podoplanin Under Flow is Mediated by the Receptor CLEC-2 and Stabilised by Src/Syk-Dependent Platelet Signalling
Identifieur interne : 001987 ( Main/Exploration ); précédent : 001986; suivant : 001988Platelet Adhesion to Podoplanin Under Flow is Mediated by the Receptor CLEC-2 and Stabilised by Src/Syk-Dependent Platelet Signalling
Auteurs : Leyre Navarro-Nú Ez ; Alice Y. Pollitt ; Kate Lowe ; Arusa Latif ; Gerard B. Nash ; Steve P. WatsonSource :
- Thrombosis and Haemostasis [ 0340-6245 ] ; 2015.
Abstract
Platelet-specific deletion of CLEC-2, which signals through Src and Syk kinases, or global deletion of its ligand podoplanin results in blood-filled lymphatics during mouse development. Platelet-specific Syk deficiency phenocopies this defect, indicating that platelet activation is required for lymphatic development. In the present study, we investigated whether CLEC-2-podoplanin interactions could support platelet arrest from blood flow and whether platelet signalling is required for stable platelet adhesion to lymphatic endothelial cells (LECs) and recombinant podoplanin under flow. Perfusion of human or mouse blood over human LEC monolayers led to platelet adhesion and aggregation. Following αIIbβ3 blockade, individual platelets still adhered. Platelet binding occurred at venous but not arterial shear rates. There was no adhesion using CLEC-2-deficient blood or to vascular endothelial cells (which lack podoplanin). Perfusion of human blood over human Fc-podoplanin (hFcPDPN) in the presence of monoclonal antibody IV.3 to block FcγRIIA receptors led to platelet arrest at similar shear rates to those used on LECs. Src and Syk inhibitors significantly reduced global adhesion of human or mouse platelets to LECs and hFcPDPN. A similar result was seen using Syk-deficient mouse platelets. Reduced platelet adhesion was due to a decrease in the stability of binding. In conclusion, our data reveal that CLEC-2 is an adhesive receptor that supports platelet arrest to podoplanin under venous shear. Src/Syk-dependent signalling stabilises platelet adhesion to podoplanin, providing a possible molecular mechanism contributing to the lymphatic defects of Syk-deficient mice.
Url:
DOI: 10.1160/TH14-09-0762
PubMed: 25694214
PubMed Central: 4990172
Affiliations:
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Nash</name></author><author><name sortKey="Watson, Steve P" sort="Watson, Steve P" uniqKey="Watson S" first="Steve P." last="Watson">Steve P. 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Pollitt</name></author><author><name sortKey="Lowe, Kate" sort="Lowe, Kate" uniqKey="Lowe K" first="Kate" last="Lowe">Kate Lowe</name></author><author><name sortKey="Latif, Arusa" sort="Latif, Arusa" uniqKey="Latif A" first="Arusa" last="Latif">Arusa Latif</name></author><author><name sortKey="Nash, Gerard B" sort="Nash, Gerard B" uniqKey="Nash G" first="Gerard B." last="Nash">Gerard B. Nash</name></author><author><name sortKey="Watson, Steve P" sort="Watson, Steve P" uniqKey="Watson S" first="Steve P." last="Watson">Steve P. Watson</name></author></analytic><series><title level="j">Thrombosis and Haemostasis</title><idno type="ISSN">0340-6245</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>Summary</title><p>Platelet-specific deletion of CLEC-2, which signals through Src and Syk kinases, or global deletion of its ligand podoplanin results in blood-filled lymphatics during mouse development. Platelet-specific Syk deficiency phenocopies this defect, indicating that platelet activation is required for lymphatic development. In the present study, we investigated whether CLEC-2-podoplanin interactions could support platelet arrest from blood flow and whether platelet signalling is required for stable platelet adhesion to lymphatic endothelial cells (LECs) and recombinant podoplanin under flow. Perfusion of human or mouse blood over human LEC monolayers led to platelet adhesion and aggregation. Following αIIbβ3 blockade, individual platelets still adhered. Platelet binding occurred at venous but not arterial shear rates. There was no adhesion using CLEC-2-deficient blood or to vascular endothelial cells (which lack podoplanin). Perfusion of human blood over human Fc-podoplanin (hFcPDPN) in the presence of monoclonal antibody IV.3 to block FcγRIIA receptors led to platelet arrest at similar shear rates to those used on LECs. Src and Syk inhibitors significantly reduced global adhesion of human or mouse platelets to LECs and hFcPDPN. A similar result was seen using Syk-deficient mouse platelets. Reduced platelet adhesion was due to a decrease in the stability of binding. In conclusion, our data reveal that CLEC-2 is an adhesive receptor that supports platelet arrest to podoplanin under venous shear. Src/Syk-dependent signalling stabilises platelet adhesion to podoplanin, providing a possible molecular mechanism contributing to the lymphatic defects of Syk-deficient mice.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="H Gerling, R" uniqKey="H Gerling R">R Hägerling</name></author><author><name sortKey="Pollmann, C" uniqKey="Pollmann C">C Pollmann</name></author><author><name sortKey="Andreas, M" uniqKey="Andreas M">M Andreas</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yang, Y" uniqKey="Yang Y">Y Yang</name></author><author><name sortKey="Garcia Verdugo, Jm" uniqKey="Garcia Verdugo J">JM Garcia-Verdugo</name></author><author><name sortKey="Soriano Navarro, M" uniqKey="Soriano Navarro M">M Soriano-Navarro</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tober, J" uniqKey="Tober J">J Tober</name></author><author><name sortKey="Koniski, A" uniqKey="Koniski 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Verzeroli</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Konstantopoulos, K" uniqKey="Konstantopoulos K">K Konstantopoulos</name></author><author><name sortKey="Thomas, Sn" uniqKey="Thomas S">SN Thomas</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cueni, Ln" uniqKey="Cueni L">LN Cueni</name></author><author><name sortKey="Hegyi, I" uniqKey="Hegyi I">I Hegyi</name></author><author><name sortKey="Shin, Jw" uniqKey="Shin J">JW Shin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kunita, A" uniqKey="Kunita A">A Kunita</name></author><author><name sortKey="Kashima, Tg" uniqKey="Kashima T">TG Kashima</name></author><author><name sortKey="Morishita, Y" uniqKey="Morishita Y">Y Morishita</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kerrigan, Am" uniqKey="Kerrigan A">AM Kerrigan</name></author><author><name sortKey="Navarro Nu Ez, L" uniqKey="Navarro Nu Ez L">L Navarro-Núñez</name></author><author><name sortKey="Pyz, E" uniqKey="Pyz E">E Pyz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hatakeyama, K" uniqKey="Hatakeyama K">K Hatakeyama</name></author><author><name sortKey="Kaneko, Mk" uniqKey="Kaneko M">MK Kaneko</name></author><author><name sortKey="Kato, Y" uniqKey="Kato Y">Y Kato</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list></list><tree><noCountry><name sortKey="Latif, Arusa" sort="Latif, Arusa" uniqKey="Latif A" first="Arusa" last="Latif">Arusa Latif</name><name sortKey="Lowe, Kate" sort="Lowe, Kate" uniqKey="Lowe K" first="Kate" last="Lowe">Kate Lowe</name><name sortKey="Nash, Gerard B" sort="Nash, Gerard B" uniqKey="Nash G" first="Gerard B." last="Nash">Gerard B. 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