Deficiency of platelet adhesion molecule CD226 causes megakaryocyte development and platelet hyperactivity

Abstract

This study used constitutive CD226 gene knockout (KO) mice as a model to investigate the functions and mechanisms of CD226 in megakaryocyte (MK) maturation and platelet activation. Although CD226 deficiency did not cause MK polyploidization or platelet granule abnormalities, increased MK counts were detected in the femora bone marrow (BM) and spleen of CD226 KO mice. Particularly, CD226 KO mice have a more extensive membrane system in MKs and platelets than wild-type (WT) mice. We also demonstrated that CD226 KO mice displayed increased platelet counts, shortened bleeding time, and enhanced platelet aggregation. CD226 KO platelets had an increased mature platelet ratio compared to the control platelets. In addition, the observed reduction in bleeding time may be due to decreased nitric oxide (NO) production in the platelets. Platelet-specific CD226-deficient mice showed similar increased MK counts, shortened bleeding time, enhanced platelet aggregation, and decreased NO production in platelets. Furthermore, we performed middle cerebral artery occlusion-reperfusion surgery on WT and CD226 KO mice to explore the potential effect of CD226 on acute ischemia-reperfusion injury; the results revealed that CD226 deficiency led to significantly increased infarct area. Thus, CD226 is a promising candidate for the treatment of thrombotic disorders. K E Y W O R D Sactivation, CD226, megakaryocyte, nitric oxide, platelet 6872 | ZHANG et Al.

CD226 Is Required to Maintain Megakaryocytes/Platelets Homeostasis in the Treatment of Knee Osteoarthritis With Platelet-Rich Plasma in Mice

“…Further study indicated that the deletion of CD226 resulted in mild thrombocytosis and increased MK/platelet number. Platelet hyperactivity was partially due to the decreased platelet-derived nitroxide level in CD226 global knockout mice ( Zhang et al, 2020 ). These findings indicate that CD226 is a potential regulator for intravascular platelet activation to be regarded as antithrombotic agents.…”

“…PRP was separated at the upper layer from the blood samples by centrifugation at room temperature (RT) for 10 min at 160 g/min. For the preparation of washed platelets, PRP was centrifuged at 2,000 g/min for 10 min at RT, and the platelet pellet was resuspended ( Zhang et al, 2020 ).…”

“…Platelets stained with APC-CD41 antibodies (#17–0,411-82, eBioscience) were distinguished from other blood cells. Subsequently, blood cells were incubated with 1 μg/ml thiazole orange (TO; Sigma-Aldrich) dye for 15 min at RT to stain immature reticulated platelets and fixed with formaldehyde, followed by three-color flow cytometric analysis to determine the immature/mature percentage of platelets from day 1 to day 5 as previously reported ( Zhang et al, 2020 ).…”

Identification of lncRNA–mRNA Regulatory Network Associated With Isolated Systolic Hypertension And Atherosclerotic Cerebral Infarction

“…CD226 was expressed on the platelet membrane and was involved in platelet activation [11,12]. Platelets played a crucial role in the control of acute hemorrhage and thrombosis, and thus were involved in atherosclerosis and myocardial infarction [13]. CD226 may participate in atherosclerotic cerebral infarction.…”

Identification of lncRNA–mRNA Regulatory Network Associated With Isolated Systolic Hypertension and Atherosclerotic Cerebral Infarction

“…CD226 was expressed on the platelet membrane and was involved in platelet activation [11,12]. Platelets played a crucial role in the control of acute hemorrhage and thrombosis, and thus were involved in atherosclerosis and myocardial infarction [13]. CD226 may participate in atherosclerotic cerebral infarction.…”