TY - JOUR
T1 - Nonredundant Roles of Platelet Glycoprotein VI and Integrin αIIbβ3 in Fibrin-Mediated Microthrombus Formation
AU - Perrella, Gina
AU - Huang, Jingnan
AU - Provenzale, Isabella
AU - Swieringa, Frauke
AU - Heubel-Moenen, Floor C. J.
AU - Farndale, Richard W.
AU - Roest, Mark
AU - van der Meijden, Paola E. J.
AU - Thomas, Mark
AU - Ariens, Robert A. S.
AU - Jandrot-Perrus, Martine
AU - Watson, Steve P.
AU - Heemskerk, Johan W. M.
N1 - Funding Information:
G. Perrella is supported by a joint PhD scholarship of Maastricht and Birmingham Universities. J. Huang and I. Provenzale are supported by the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No. 766118, and are registered in the PhD programs of Maastricht and Santiago da Compostela Universities (J. Huang) or Maastricht and Reading Universities (I. Provenzale). S.P. Watson and R.A.S. Ariëns are supported by a Wellcome Trust Joint Investigator Award (204951/Z/16/Z). S.P. Watson holds a British Heart Foundation Chair (CH/03/003).
Publisher Copyright:
© 2020 American Heart Association, Inc.
PY - 2021/2
Y1 - 2021/2
N2 - Objective:Fibrin is considered to strengthen thrombus formation via integrin alpha IIb beta 3, but recent findings indicate that fibrin can also act as ligand for platelet glycoprotein VI.Approach and Results:To investigate the thrombus-forming potential of fibrin and the roles of platelet receptors herein, we generated a range of immobilized fibrin surfaces, some of which were cross-linked with factor XIIIa and contained VWF-BP (von Willebrand factor-binding peptide). Multicolor microfluidics assays with whole-blood flowed at high shear rate (1000 s(-1)) indicated that the fibrin surfaces, regardless of the presence of factor XIIIa or VWF-BP, supported platelet adhesion and activation (P-selectin expression), but only microthrombi were formed consisting of bilayers of platelets. Fibrinogen surfaces produced similar microthrombi. Markedly, tiggering of coagulation with tissue factor or blocking of thrombin no more than moderately affected the fibrin-induced microthrombus formation. Absence of alpha IIb beta 3 in Glanzmann thrombasthenia annulled platelet adhesion. Blocking of glycoprotein VI with Fab 9O12 substantially, but incompletely reduced platelet secretion, Ca2+ signaling and aggregation, while inhibition of Syk further reduced these responses. In platelet suspension, glycoprotein VI blockage or Syk inhibition prevented fibrin-induced platelet aggregation. Microthrombi on fibrin surfaces triggered only minimal thrombin generation, in spite of thrombin binding to the fibrin fibers.Conclusions:Together, these results indicate that fibrin fibers, regardless of their way of formation, act as a consolidating surface in microthrombus formation via nonredundant roles of platelet glycoprotein VI and integrin alpha IIb beta 3 through signaling via Syk and low-level Ca2+ rises.
AB - Objective:Fibrin is considered to strengthen thrombus formation via integrin alpha IIb beta 3, but recent findings indicate that fibrin can also act as ligand for platelet glycoprotein VI.Approach and Results:To investigate the thrombus-forming potential of fibrin and the roles of platelet receptors herein, we generated a range of immobilized fibrin surfaces, some of which were cross-linked with factor XIIIa and contained VWF-BP (von Willebrand factor-binding peptide). Multicolor microfluidics assays with whole-blood flowed at high shear rate (1000 s(-1)) indicated that the fibrin surfaces, regardless of the presence of factor XIIIa or VWF-BP, supported platelet adhesion and activation (P-selectin expression), but only microthrombi were formed consisting of bilayers of platelets. Fibrinogen surfaces produced similar microthrombi. Markedly, tiggering of coagulation with tissue factor or blocking of thrombin no more than moderately affected the fibrin-induced microthrombus formation. Absence of alpha IIb beta 3 in Glanzmann thrombasthenia annulled platelet adhesion. Blocking of glycoprotein VI with Fab 9O12 substantially, but incompletely reduced platelet secretion, Ca2+ signaling and aggregation, while inhibition of Syk further reduced these responses. In platelet suspension, glycoprotein VI blockage or Syk inhibition prevented fibrin-induced platelet aggregation. Microthrombi on fibrin surfaces triggered only minimal thrombin generation, in spite of thrombin binding to the fibrin fibers.Conclusions:Together, these results indicate that fibrin fibers, regardless of their way of formation, act as a consolidating surface in microthrombus formation via nonredundant roles of platelet glycoprotein VI and integrin alpha IIb beta 3 through signaling via Syk and low-level Ca2+ rises.
KW - blood platelet
KW - fibrin
KW - microfluidics
KW - platelet aggregation
KW - thrombin
KW - VON-WILLEBRAND-FACTOR
KW - THROMBUS FORMATION
KW - PROCOAGULANT ACTIVITY
KW - TISSUE FACTOR
KW - KEY ROLE
KW - COLLAGEN
KW - KINASE
KW - RECEPTOR
KW - GPVI
KW - ADHESION
U2 - 10.1161/ATVBAHA.120.314641
DO - 10.1161/ATVBAHA.120.314641
M3 - Article
C2 - 33267658
SN - 1079-5642
VL - 41
SP - E97-E111
JO - Arteriosclerosis Thrombosis and Vascular Biology
JF - Arteriosclerosis Thrombosis and Vascular Biology
IS - 2
ER -