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

Laura Delvasto-Núñez; Dorina Roem; Kamran Bakhtiari; Gerard van Mierlo; Joost C. M. Meijers; Ilse Jongerius; Sacha S. Zeerleder

doi:10.1055/a-1497-9573

Thrombosis and Haemostasis, Table of Contents

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

¹Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands

²Department of Hematology, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands

,

Dorina Roem

¹Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands

,

Kamran Bakhtiari

³Department of Molecular Hematology, Sanquin Research, Amsterdam, The Netherlands

,

Gerard van Mierlo

¹Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands

,

Joost C. M. Meijers

³Department of Molecular Hematology, Sanquin Research, Amsterdam, The Netherlands

⁴Department of Experimental Vascular Medicine, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands

,

Ilse Jongerius ‡^*

¹Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands

⁵Department of Pediatric Immunology, Rheumatology and Infectious Diseases, Emma Children's Hospital, Amsterdam UMC, Amsterdam, the Netherlands

,

Sacha S. Zeerleder‡^*

¹Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands

²Department of Hematology, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands

⁶Department of Hematology and Central Hematology Laboratory, Inselspital – Bern University Hospital, University of Bern, Bern, Switzerland

⁷Department for BioMedical Research, University of Bern, Bern, Switzerland

› Author Affiliations

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

Full Text

References

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Supplementary Material