Inhibition of Hypoxia-Inducible Factor Prolyl-Hydroxylase Modulates Platelet Function

Wei Gu; Jiaqian Qi; Sixuan Zhang; Yangyang Ding; Jianlin Qiao; Yue Han

doi:10.1055/a-1837-7797

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2022; 122(10): 1693-1705
DOI: 10.1055/a-1837-7797

Cellular Haemostasis and Platelets

Inhibition of Hypoxia-Inducible Factor Prolyl-Hydroxylase Modulates Platelet Function

Wei Gu

¹National Clinical Research Center for Hematologic Diseases, The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou, China

,

Jiaqian Qi

¹National Clinical Research Center for Hematologic Diseases, The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou, China

²Collaborative Innovation Center of Hematology, Institute of Blood and Marrow Transplantation, Soochow University, Suzhou, China

³Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou, China

⁴State Key Laboratory of Radiation Medicine and Protection, Soochow University, Suzhou, China

,

Sixuan Zhang

⁵Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China

,

Yangyang Ding

⁵Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China

,

Jianlin Qiao

⁵Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China

,

Yue Han

¹National Clinical Research Center for Hematologic Diseases, The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou, China

²Collaborative Innovation Center of Hematology, Institute of Blood and Marrow Transplantation, Soochow University, Suzhou, China

³Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou, China

⁴State Key Laboratory of Radiation Medicine and Protection, Soochow University, Suzhou, China

› Author Affiliations

Abstract

Hypoxia-inducible factor-1α (HIF-1α) involves in redox reactions. Considering the role of reactive oxygen species (ROS) in platelet function, whether it regulates platelet function remains unclear. Using an inhibitor of HIF prolyl–hydroxylase, IOX-2, we intend to investigate its effect on platelet function. Human platelets were treated with IOX-2 (0, 10, 25, and 50 μM) followed by analysis of platelet aggregation, granule secretion, receptor expression, platelet spreading, or clot retraction. Additionally, IOX-2 (10 mg/kg) was injected intraperitoneally into mice to measure tail bleeding time and arterial thrombosis. IOX-2 significantly inhibited collagen-related peptide (CRP; 0.25 μg/mL) or thrombin (0.03 U/mL)-induced platelet aggregation and ATP release dose dependently without affecting P-selectin expression and the surface levels of glycoprotein (GP)Ibα, GPVI, or α_IIbβ₃. In addition, IOX-2-treated platelets presented significantly decreased spreading on fibrinogen or collagen and clot retraction. Moreover, IOX-2 administration into mice significantly impaired the in vivo hemostatic function of platelets and arterial thrombus formation without affecting the number of circulating platelets and coagulation factors (FVIII and FIX). Further, IOX-2 significantly upregulated HIF-1α in platelets, decreased ROS generation, and downregulated NOX1 expression. Finally, IOX-2 increased the phosphorylation level of VASP (Ser157/239), and inhibited the phosphorylation of p38 (Thr180/Tyr182), ERK1/2 (Thr202/Tyr204), AKT (Thr308/Ser473), and PKCδ (Thr505) in CRP- or thrombin-stimulated platelets. In conclusion, inhibition of HIF prolyl-hydroxylase modulates platelet function and arterial thrombus formation, possibly through upregulation of HIF-1α expression and subsequent inhibition of ROS generation, indicating that HIF-1α might be a novel target for the treatment of thrombotic disorders.

Keywords

HIF-1α - IOX-2 - platelet function - hemostasis - arterial thrombosis - ROS

Full Text

References

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Supplementary Material