Inhibition of IP3 (Inositol 1,4,5-Trisphosphate) Receptors Retards SARS-CoV-2-Induced Endothelial von Willebrand Factor Secretion and Thrombosis

Xin-yi Yu; Xin-yu Jia; Ting-yu Wang; Yan-hong Zhang; Hao Song; Kan Li; Zhuo-zheng Chen; Yi Zhu; Liu Yao

doi:10.1055/a-2471-8767

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost
DOI: 10.1055/a-2471-8767

Coagulation and Fibrinolysis

Inhibition of IP3 (Inositol 1,4,5-Trisphosphate) Receptors Retards SARS-CoV-2-Induced Endothelial von Willebrand Factor Secretion and Thrombosis

Xin-yi Yu

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Xin-yu Jia

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Ting-yu Wang

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Yan-hong Zhang

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Hao Song

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Kan Li

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Zhuo-zheng Chen

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Yi Zhu

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

,

Liu Yao

¹The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China

› Author Affiliations

Abstract

Patients with coronavirus disease 2019 (COVID-19) are at high risk of developing a hypercoagulable state and thrombosis. The von Willebrand factor (vWF) produced by endothelial cells (ECs) is a critical thrombosis regulator. We previously found that cytoskeleton-associated protein 4 (CKAP4) is a novel receptor for the spike protein of severe acute respiratory syndrome coronavirus-2 and is involved in COVID-19-associated coagulopathy. However, the underlying mechanism involved remains unclear. This study aimed to explore the signaling pathways involved in spike protein-CKAP4-induced vWF secretion and thrombosis. Treatment of ECs with the spike protein significantly induced vWF secretion, coagulation factor VIII (FVIII)–vWF binding, and platelet adhesion to ECs, which were blocked by the selective intracellular calcium chelator, BAPTA-AM. Furthermore, using several calcium channel-blocking drugs and small-molecule inhibitors, we found that calcium released from the endoplasmic reticulum (ER) is involved in this process. IP3 (inositol 1,4,5-trisphosphate) receptors (IP3Rs) inhibition ameliorated spike protein-induced vWF secretion, FVIII–vWF binding affinity, and platelet adhesion to ECs. Specifically, the knockdown of IP3R1, a crucial type of IP3Rs, reversed spike protein-induced endothelial vWF secretion, and the procoagulant state. Moreover, KT-362, an investigational and clinically relevant antihypertensive drug targeting IP3Rs-mediated calcium release, repressed spike protein-induced endothelial vWF secretion. Conversely, the IP3Rs agonist promoted endothelial vWF secretion, which was not affected by CKAP4 knockdown. In vivo treatment of endothelial-specific human CKAP4 overexpression mice with KT-362 retarded spike protein-induced vWF secretion and thrombosis. Thus, IP3Rs mediated calcium release from the ER and contributed to spike protein-induced vWF secretion and thrombosis, making them potential therapeutic targets for COVID-19-associated coagulopathy.

Keywords

spike protein - endothelium - von Willebrand factor - IP3 (inositol 1,4,5-trisphosphate) receptors - thrombosis

Full Text

References

References
1 Burn E, Duarte-Salles T, Fernandez-Bertolin S. et al. Venous or arterial thrombosis and deaths among COVID-19 cases: a European network cohort study. Lancet Infect Dis 2022; 22 (08) 1142-1152
2 Malas MB, Naazie IN, Elsayed N, Mathlouthi A, Marmor R, Clary B. Thromboembolism risk of COVID-19 is high and associated with a higher risk of mortality: a systematic review and meta-analysis. EClinicalMedicine 2020; 29: 100639
3 Neubauer K, Zieger B. Endothelial cells and coagulation. Cell Tissue Res 2022; 387 (03) 391-398
4 Li K, Yao L, Wang J. et al. SARS-CoV-2 spike protein promotes vWF secretion and thrombosis via endothelial cytoskeleton-associated protein 4 (CKAP4). Signal Transduct Target Ther 2022; 7 (01) 332
5 Ackermann M, Verleden SE, Kuehnel M. et al. Pulmonary vascular endothelialitis, thrombosis, and angiogenesis in COVID-19. N Engl J Med 2020; 383 (02) 120-128
6 Goshua G, Pine AB, Meizlish ML. et al. Endotheliopathy in COVID-19-associated coagulopathy: evidence from a single-centre, cross-sectional study. Lancet Haematol 2020; 7 (08) e575-e582
7 Anderson JR, Li J, Springer TA, Brown A. Structures of VWF tubules before and after concatemerization reveal a mechanism of disulfide bond exchange. Blood 2022; 140 (12) 1419-1430
8 Haberichter SL. von Willebrand factor propeptide: biology and clinical utility. Blood 2015; 126 (15) 1753-1761
9 Philippe A, Chocron R, Gendron N. et al. Circulating von Willebrand factor and high molecular weight multimers as markers of endothelial injury predict COVID-19 in-hospital mortality. Angiogenesis 2021; 24 (03) 505-517
10 Rondaij MG, Bierings R, Kragt A, van Mourik JA, Voorberg J. Dynamics and plasticity of Weibel-Palade bodies in endothelial cells. Arterioscler Thromb Vasc Biol 2006; 26 (05) 1002-1007
11 Chen X, Cao R, Zhong W. Host calcium channels and pumps in viral infections. Cells 2019; 9 (01) 9
12 Xu J, Yang Y, Hou Z, Jia H, Wang Y. TRPV2-spike protein interaction mediates the entry of SARS-CoV-2 into macrophages in febrile conditions. Theranostics 2021; 11 (15) 7379-7390
13 Yang K, Liu S, Yan H. et al. SARS-CoV-2 spike protein receptor-binding domain perturbates intracellular calcium homeostasis and impairs pulmonary vascular endothelial cells. Signal Transduct Target Ther 2023; 8 (01) 276
14 Yao L, He J, Li B. et al. Regulation of YAP by mammalian target of rapamycin complex 1 in endothelial cells controls blood pressure through COX-2/mPGES-1/PGE(2) cascade. Hypertension 2019; 74 (04) 936-946
15 Bendetowicz AV, Morris JA, Wise RJ, Gilbert GE, Kaufman RJ. Binding of factor VIII to von Willebrand factor is enabled by cleavage of the von Willebrand factor propeptide and enhanced by formation of disulfide-linked multimers. Blood 1998; 92 (02) 529-538
16 Han X, Li P, Yang Z. et al. Zyxin regulates endothelial von Willebrand factor secretion by reorganizing actin filaments around exocytic granules. Nat Commun 2017; 8: 14639
17 Rayner SG, Scholl Z, Mandrycky CJ. et al. Endothelial-derived von Willebrand factor accelerates fibrin clotting within engineered microvessels. J Thromb Haemost 2022; 20 (07) 1627-1637
18 Ham SJ, Yoo H, Woo D, Lee DH, Park KS, Chung J. PINK1 and Parkin regulate IP₃R-mediated ER calcium release. Nat Commun 2023; 14 (01) 5202
19 Woll KA, Van Petegem F. Calcium-release channels: structure and function of IP₃ receptors and ryanodine receptors. Physiol Rev 2022; 102 (01) 209-268
20 Lin Q, Zhao L, Jing R. et al. Inositol 1,4,5-trisphosphate receptors in endothelial cells play an essential role in vasodilation and blood pressure regulation. J Am Heart Assoc 2019; 8 (04) e011704
21 Eid AH, El-Yazbi AF, Zouein F. et al. Inositol 1,4,5-trisphosphate receptors in hypertension. Front Physiol 2018; 9: 1018
22 Gupta A, Madhavan MV, Sehgal K. et al. Extrapulmonary manifestations of COVID-19. Nat Med 2020; 26 (07) 1017-1032
23 Turner S, Khan MA, Putrino D, Woodcock A, Kell DB, Pretorius E. Long COVID: pathophysiological factors and abnormalities of coagulation. Trends Endocrinol Metab 2023; 34 (06) 321-344
24 Li P, Wei G, Cao Y. et al. Myosin IIa is critical for cAMP-mediated endothelial secretion of von Willebrand factor. Blood 2018; 131 (06) 686-698
25 Xiong Y, Hu Z, Han X. et al. Hypertensive stretch regulates endothelial exocytosis of Weibel-Palade bodies through VEGF receptor 2 signaling pathways. Cell Res 2013; 23 (06) 820-834
26 Dalal PJ, Muller WA, Sullivan DP. Endothelial cell calcium signaling during barrier function and inflammation. Am J Pathol 2020; 190 (03) 535-542
27 Schmitz EA, Takahashi H, Karakas E. Structural basis for activation and gating of IP₃ receptors. Nat Commun 2022; 13 (01) 1408
28 Hester RK, Shibata S. KT-362 related effects on intracellular calcium release and associated clinical potential: arrhythmias, myocardial ischemia, and hypertension. Cardiovasc Drugs Ther 1990; 4 (05) 1345-1354
29 Kodama I, Shibata S. Effects of KT-362, a new antiarrhythmic agent with vasodilating action on intracellular calcium mobilization of atrial muscle. J Pharmacol Exp Ther 1991; 258 (01) 332-338

Supplementary Material

Supplementary Material