How To Test For Platelet Function

Transcription

1 Platelet function in thrombocytopenia Mark Roest Synapse Research Institute, Maastricht

2 No Disclosures

3 Trauma

4 1. Adhesion

5 2. Activation and Aggregation

6 3. Stable Aggregate Formation

7 4. Repair

8 Platelet Disorder Mucocutaneous bleeding low platelet count and/or dysfunction epistaxis Prolonged bleeding Heavy Bruising Petechiae Gastro-intestinal bleeding Oral bleeding Bleeding after tooth extraction Menorrhagia

9 Thrombocytopenia: Current guidelines Platelet counts < 10*10 9 platelets / L transfusion Platelet counts > 10*10 9 platelets / L no transfusion We need platelet function testing in addition to platelet counts Those patients don t need transfusion if the function of the remaining platelets is still adequate Those patients need transfusion if the function of the remaining platelets is not adequate

10 Platelet function monitoring in diagnostic settings Bleeding time PFA-100 ROTEM TEG verifynow multiplate LTA These tests are based on platelet aggregation or platelet deposition This is highly dependent on platelete concentration

11 Platelet function tests Bleeding Time and PFA: Only for VW diseases and platelet adhesion ROTEM: Only for prediction post surgery bleeding? VASP: Only for P2Y12 inhibitor monitoring VerifyNOW: Only for P2Y12 inhibitors and aspirin monitoring Multiplate: Potentially interesting no progression Aggregometry: Gold standard disappearing Whole blood platelet function flow cytometry

12 SSC Count Count Platelet Function Agonist Anti-P-selectin P-selectine αiibβ Anti-fibrinogeen FSC CD62P-PE CD62P-PE

13 Platelet Activation Test (PACT) TRAP ADP Thrombin receptor (PAR1) ADP receptor (P 2 Y 12 ) Blood Platelet Collagen receptor (GPVI) CRP P-selectin (MFI) Flow Cytometry ADP ( TRAP ( M) CRP(ng/mL) P-selectin (MFI) Master Mix P-selectin (MFI)

14 Platelet Activation Test (PACT) 1. Platelet function in stored concentrates 2. Platelet function in blood of patients with thrombocytopenia

15 Platelet Function in stored concentrates (1) P-selectin expression Day Time Days Middelburg RA, Roest M, Ham J, Coccoris M, Zwaginga JJ, van der Meer PF. Flow cytometric assessment of agonist-induced P-selectin expression as a measure of platelet quality in stored platelet concentrates. Transfusion Aug;53(8):

16 Platelet Function in stored concentrates (2) Time Days Cécile H. Kicken, Mark Roest, Yvonne M. Henskens, Bas de Laat, Dana Huskens. Monitoring platelet concentrates, submited

17 Platelet Function in stored concentrates (3) Time Days P-selectin expression (AU) SFLLRN Day a IIb b 3 activation (AU) SFLLRN Day P-selectin expression (AU) CRP Day a IIb b 3 activation (AU) CRP Day P-selectin expression (AU) ADP Day a IIb b 3 activation (AU) ADP Day P-selectin expression (AU) U Day a IIb b 3 activation (AU) U Day Roest M, van Holten TC, Fleurke GJ, Remijn JA. Platelet Activation Test in Unprocessed Blood (Pac-t-UB) to Monitor Platelet Concentrates and Whole Blood of Thrombocytopenic Patients. Transfus Med Hemother Apr;40(2):

18 Platelet Activation Test (PACT) 1. Platelet function in stored concentrates 2. Platelet function in blood of patients with thrombocytopenia

19 Platelet function in blood of patients with thrombocytopenia Healthy controls (n=10) Patients with thrombocytopenia (n=11) Bikker A, Bouman E, Sebastian S, Korporaal SJ, Urbanus RT, Fijnheer R, Boven LA, Roest M. Functional recovery of stored platelets after transfusion. Transfusion May;56(5):

20 P -s e le c tin e x p re s s io n (% ) P A R - 1 P A R - 4 C R P A D P P la te le t c o u n t (x /L ) Birol Batman, Esther R van Bladel, Mariska van Hamersveld, Pieternel CM Pasker-de Jong, Suzanne JA Korporaal, Rolf T Urbanus, Mark Roest, Leonie A Boven, Rob Fijnheer. Agonist induced platelet reactivity predicts bleeding in hemato-oncological patient, submited

21 Platelet function pre- and post-transfusion Bikker A, Bouman E, Sebastian S, Korporaal SJ, Urbanus RT, Fijnheer R, Boven LA, Roest M. Functional recovery of stored platelets after transfusion. Transfusion May;56(5):

22 M F I P -s e le c tin (u n its ) Platelet function, platelet count and bleeding risk Platelet function and bleeding Platelet count and bleeding P A R 1 -A P P A R 4 -A P * M F I P -s e le c tin (u n its ) ** 0 0 N o B le e d in g B le e d in g N o B le e d in g B le e d in g Birol Batman, Esther R van Bladel, Mariska van Hamersveld, Pieternel CM Pasker-de Jong, Suzanne JA Korporaal, Rolf T Urbanus, Mark Roest, Leonie A Boven, Rob Fijnheer. Agonist induced platelet reactivity predicts bleeding in hemato-oncological patient, submited

23 ITP Auto-antibodies to platelets Increased Platelet Consumption Low Platelet Counts ITP has a heterogeneous pathogenesis and the phenotype does not necessarily correlate with platelet counts Platelet Function Testing to customize the therapy?

24 Platelet function testing in ITP van Bladel, Laarhoven AG, van der Heijden LB, Heitink-Pollé KM, Porcelijn L, van der Schoot CE, de Haas M, Roest M, Vidarsson G, de Groot PG, Bruin MCFunctional platelet defects in children with severe chronic ITP as tested with 2 novel assays applicable for low platelet counts. Blood 2014;123:

25 Conclusion PACT can be used to monitor platelet function in concentrates PACT can be used to measure platelet function in thrombocytopenia Upcoming studies will show if this test can be used to customize the treatment of patients with thrombocytopenia

26 Mark Roest Philip de Groot Dana Huskens Bas de Laat Rob Fijnheer Leonie Boven Esther van Bladel Birol Batman Rolf Urbanus Suzanne Korporaal Albert Huisman Angela Bikker Jaap Jan Zwaginga Rutger Middelburg Jasper Remijn