TY - JOUR
T1 - Platelet-primed interactions of coagulation and anticoagulation pathways in flow-dependent thrombus formation
AU - Brouns, Sanne L N
AU - van Geffen, Johanna P
AU - Campello, Elena
AU - Swieringa, Frauke
AU - Spiezia, Luca
AU - van Oerle, René
AU - Provenzale, Isabella
AU - Verdoold, Remco
AU - Farndale, Richard W
AU - Clemetson, Kenneth J
AU - Spronk, Henri M H
AU - van der Meijden, Paola E J
AU - Cavill, Rachel
AU - Kuijpers, Marijke J E
AU - Castoldi, Elisabetta
AU - Simioni, Paolo
AU - Heemskerk, Johan W M
N1 - Publisher Copyright:
© 2020, The Author(s).
PY - 2020/7/17
Y1 - 2020/7/17
N2 - In haemostasis and thrombosis, platelet, coagulation and anticoagulation pathways act together to produce fibrin-containing thrombi. We developed a microspot-based technique, in which we assessed platelet adhesion, platelet activation, thrombus structure and fibrin clot formation in real time using flowing whole blood. Microspots were made from distinct platelet-adhesive surfaces in the absence or presence of tissue factor, thrombomodulin or activated protein C. Kinetics of platelet activation, thrombus structure and fibrin formation were assessed by fluorescence microscopy. This work revealed: (1) a priming role of platelet adhesion in thrombus contraction and subsequent fibrin formation; (2) a surface-independent role of tissue factor, independent of the shear rate; (3) a mechanism of tissue factor-enhanced activation of the intrinsic coagulation pathway; (4) a local, suppressive role of the anticoagulant thrombomodulin/protein C pathway under flow. Multiparameter analysis using blood samples from patients with (anti)coagulation disorders indicated characteristic defects in thrombus formation, in cases of factor V, XI or XII deficiency; and in contrast, thrombogenic effects in patients with factor V-Leiden. Taken together, this integrative phenotyping approach of platelet-fibrin thrombus formation has revealed interaction mechanisms of platelet-primed key haemostatic pathways with alterations in patients with (anti)coagulation defects. It can help as an important functional add-on whole-blood phenotyping.
AB - In haemostasis and thrombosis, platelet, coagulation and anticoagulation pathways act together to produce fibrin-containing thrombi. We developed a microspot-based technique, in which we assessed platelet adhesion, platelet activation, thrombus structure and fibrin clot formation in real time using flowing whole blood. Microspots were made from distinct platelet-adhesive surfaces in the absence or presence of tissue factor, thrombomodulin or activated protein C. Kinetics of platelet activation, thrombus structure and fibrin formation were assessed by fluorescence microscopy. This work revealed: (1) a priming role of platelet adhesion in thrombus contraction and subsequent fibrin formation; (2) a surface-independent role of tissue factor, independent of the shear rate; (3) a mechanism of tissue factor-enhanced activation of the intrinsic coagulation pathway; (4) a local, suppressive role of the anticoagulant thrombomodulin/protein C pathway under flow. Multiparameter analysis using blood samples from patients with (anti)coagulation disorders indicated characteristic defects in thrombus formation, in cases of factor V, XI or XII deficiency; and in contrast, thrombogenic effects in patients with factor V-Leiden. Taken together, this integrative phenotyping approach of platelet-fibrin thrombus formation has revealed interaction mechanisms of platelet-primed key haemostatic pathways with alterations in patients with (anti)coagulation defects. It can help as an important functional add-on whole-blood phenotyping.
KW - ACTIVATION
KW - BLOOD-COAGULATION
KW - COLLAGEN
KW - CONCENTRATE
KW - FIBRIN
KW - GENERATION
KW - HEMOSTASIS
KW - PLASMA
KW - PROTEIN-C
KW - RESISTANCE
U2 - 10.1038/s41598-020-68438-9
DO - 10.1038/s41598-020-68438-9
M3 - Article
C2 - 32680988
SN - 2045-2322
VL - 10
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 11910
ER -