Selective inhibition of the platelet phosphoinositide 3-kinase p110beta as promising new strategy for platelet protection during extracorporeal circulation.
Extracorporeal circulation (ECC) is used in cardiac surgery for cardiopulmonary bypass as well as in ventricular assist devices and for extracorporeal membrane oxygenation. Blood contact with the artificial surface and shear stress of ECC activates platelets and leukocytes resulting in a coagulopathy and proinflammatory events. Blockers of the platelet glycoprotein (GP) IIb/IIIa (CD41/CD61) can protect platelet function during ECC, a phenomenon called "platelet anaesthesia", but may be involved in post-ECC bleeding. We hypothesized that the new selective phosphoinositide 3-kinase p110beta inhibitor TGX-221 that inhibits shear-induced platelet activation without prolonging the bleeding time in vivo may also protect platelet function during ECC. Heparinized blood of healthy volunteers (n = 6) was treated in vitro with either the GP IIb/IIIa blocker tirofiban, TGX-221 or as control and circulated in an ECC model. Before and after 30 minutes circulation CD41 expression on the ECC-tubing as measure for platelet-ECC binding and generation of the platelet activation marker beta-thromboglobulin were determined using ELISA. Platelet aggregation and platelet-granulocyte binding were analysed in flow cytometry. After log-transforming the data statistical evaluation was performed using multifactor ANOVA in combination with Tukey's HSD test (global alpha = 5%). Tirofiban and TGX-221 inhibited platelet-ECC interaction, platelet aggregation and platelet-granulocyte binding. Tirofiban also inhibited ECC-induced beta-thromboglobulin release. The observed inhibition of platelet-ECC interaction and platelet activation by tirofiban contributes to explain the mechanism of "platelet anaesthesia". TGX-221 represents a promising alternative to GP IIb/IIIa blockade and should be further investigated for use during ECC in vivo.
Straub
Andreas
A
Dept. of Thoracic, Cardiac and Vascular Surgery, University of Tübingen, Germany. Andreas.Straub@baker.edu.au
Wendel
Hans Peter
HP
Dietz
Klaus
K
Schiebold
Daniela
D
Peter
Karlheinz
K
Schoenwaelder
Simone M
SM
Ziemer
Gerhard
G
D000990:Antithrombin III / Q000378:metabolism; D001792:Blood Platelets / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D058534:Class I Phosphatidylinositol 3-Kinases; D005112:Extracorporeal Circulation / Q000009:adverse effects* / Q000295:instrumentation; D005340:Fibrinogen / Q000378:metabolism; D006098:Granulocytes / Q000187:drug effects / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D008297:Male; D008855:Microscopy, Electron, Scanning; D009025:Morpholines / Q000494:pharmacology*; D010447:Peptide Hydrolases / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D015539:Platelet Activation / Q000187:drug effects*; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000037:antagonists & inhibitors / Q000378:metabolism; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011744:Pyrimidinones / Q000494:pharmacology*; D012107:Research Design; D013314:Stress, Mechanical; D013997:Time Factors; D000077466:Tirofiban; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology; D001620:beta-Thromboglobulin / Q000378:metabolism
D000990:Antithrombin III; D058534:Class I Phosphatidylinositol 3-Kinases; D005340:Fibrinogen; D006454:Hemoglobins; D009025:Morpholines; D010447:Peptide Hydrolases; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010975:Platelet Aggregation Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D040201:Platelet Membrane Glycoprotein IIb; D047428:Protein Kinase Inhibitors; D011744:Pyrimidinones; D000077466:Tirofiban; D014443:Tyrosine; D001620:beta-Thromboglobulin; C504718:TGX 221; C046193:antithrombin III-protease complex; C497132:PIK3CB protein, human