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Thromb Haemost 2022; 122(01): 080-091
DOI: 10.1055/a-1497-9573
Coagulation and Fibrinolysis

Iron-Driven Alterations on Red Blood Cell-Derived Microvesicles Amplify Coagulation during Hemolysis via the Intrinsic Tenase Complex

Laura Delvasto-Núñez
1   Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands
2   Department of Hematology, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands
,
Dorina Roem
1   Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands
,
Kamran Bakhtiari
3   Department of Molecular Hematology, Sanquin Research, Amsterdam, The Netherlands
,
Gerard van Mierlo
1   Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands
,
Joost C. M. Meijers
3   Department of Molecular Hematology, Sanquin Research, Amsterdam, The Netherlands
4   Department of Experimental Vascular Medicine, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands
,
Ilse Jongerius  ‡   *
1   Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands
5   Department of Pediatric Immunology, Rheumatology and Infectious Diseases, Emma Children's Hospital, Amsterdam UMC, Amsterdam, the Netherlands
,
Sacha S. Zeerleder‡   *
1   Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands
2   Department of Hematology, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands
6   Department of Hematology and Central Hematology Laboratory, Inselspital – Bern University Hospital, University of Bern, Bern, Switzerland
7   Department for BioMedical Research, University of Bern, Bern, Switzerland
› Author Affiliations Funding This study was supported by research funding from Trombosestichting Nederland to S.S.Z. (grant number 201604) and by a Product and Process Development grant obtained (in competition) from Sanquin Blood Supply Foundation (PPOC19–24/L2467) to I.J.
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Abstract

Hemolytic disorders characterized by complement-mediated intravascular hemolysis, such as autoimmune hemolytic anemia and paroxysmal nocturnal hemoglobinuria, are often complicated by life-threatening thromboembolic complications. Severe hemolytic episodes result in the release of red blood cell (RBC)-derived proinflammatory and oxidatively reactive mediators (e.g., extracellular hemoglobin, heme, and iron) into plasma. Here, we studied the role of these hemolytic mediators in coagulation activation by measuring factor Xa (FXa) and thrombin generation in the presence of RBC lysates. Our results show that hemolytic microvesicles (HMVs) formed during hemolysis stimulate thrombin generation through a mechanism involving FVIII and FIX, the so-called intrinsic tenase complex. Iron scavenging during hemolysis using deferoxamine decreased the ability of the HMVs to enhance thrombin generation. Furthermore, the addition of ferric chloride (FeCl3) to plasma propagated thrombin generation in a FVIII- and FIX-dependent manner suggesting that iron positively affects blood coagulation. Phosphatidylserine (PS) blockade using lactadherin and iron chelation using deferoxamine reduced intrinsic tenase activity in a purified system containing HMVs as source of phospholipids confirming that both PS and iron ions contribute to the procoagulant effect of the HMVs. Finally, the effects of FeCl3 and HMVs decreased in the presence of ascorbate and glutathione indicating that oxidative stress plays a role in hypercoagulability. Overall, our results provide evidence for the contribution of iron ions derived from hemolytic RBCs to thrombin generation. These findings add to our understanding of the pathogenesis of thrombosis in hemolytic diseases.

Keywords

coagulation - red blood cells - iron - hemolysis - intrinsic tenase complex

Author Contributions

L.D.N., J.C.M.M., I.J., and S.S.Z. conceptualized the study and contributed to the design and execution of the research. I.J. and S.S.Z. obtained funding for the study. L.D.N., D.R., and K.B. performed the experiments and analyzed the data; L.D.N., D.R., K.B., G.v.M., J.C.M.M., I.J., and S.S.Z. interpreted the data and designed the experiments. L.D.N., J.C.M.M., I.J., and S.S.Z. wrote the initial draft of the manuscript and all authors critically commented on the manuscript.


* These authors contributed equally.


Supplementary Material



Publication History

Received: 04 November 2020

Accepted: 26 April 2021

Accepted Manuscript online:
03 May 2021

Article published online:
27 September 2021

© 2021. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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