Abstract

Thrombosis are severe complications of paroxysmal nocturnal hemoglobinuria (PNH), effectively reduced by eculizumab. Extracellular vesicles (EVs) may play a central role. The objective of this study was to assess the procoagulant activity of plasma isolated from PNH patients (treated or not by eculizumab) and to quantify their circulating EVs.We iteratively collected the platelet-free-plasma of 17 PNH patients and 16 matched healthy volunteers, quantified their circulating EVs by flow cytometry and evaluated their procoagulant activity by thrombin generation and STA-Procoag-procoagulant phospholipid (PPL) assays.A significant decrease of EVs from platelets (P = .024) and an increase of the STA-Procoag-PPL clotting time (P = .049) was observed after initiation of eculizumab and up to 11 weeks after. This reduction of prothrombotic biomarkers was not observed with the thrombin generation test due to a lack of sensitivity of this assay. Active hemolysis was observed in 90% of patients and elevated D-dimers in 41% of them. However, no significant difference was observed between patients and control subjects regarding the procoagulant activity, the EVs quantity, or the cellular origin. Lactate dehydrogenase (LDH) levels were lower in eculizumab-treated patients compared to nontreated patients (441 vs 2448 IU/L). D-dimers and LDH decreased after administration of eculizumab (mean decrease of 1307 ng/mL and 4159 IU/L, respectively).These observations suggest a decrease of the phospholipid-dependent procoagulant potential of EVs after eculizumab therapy in PNH patients. TRIAL REGISTRATION:: NUB: B039201214365.

Original languageEnglish
Pages (from-to)e16164
Number of pages8
Journal Medicine (Mumbai): analytical reviews of general medicine, neurology, psychiatry, dermatology and pediatrics
Volume98
Issue number27
DOIs
Publication statusPublished - 1 Jul 2019

Fingerprint

Paroxysmal Hemoglobinuria
Blood Platelets
Prospective Studies
Phospholipids
Thrombin
Hemolysis
Extracellular Vesicles
eculizumab
L-Lactate Dehydrogenase
Healthy Volunteers
Flow Cytometry
Thrombosis
Biomarkers

Keywords

  • Administration, Intravenous
  • Adult
  • Aged
  • Antibodies, Monoclonal, Humanized/administration & dosage
  • Case-Control Studies
  • Extracellular Vesicles/drug effects
  • Flow Cytometry
  • Hemoglobinuria, Paroxysmal/blood
  • Humans
  • Middle Aged
  • Prospective Studies
  • Thrombosis/etiology

Cite this

@article{8af8109a59ba46bea494cb01562e725a,
title = "Prospective and comparative study of paroxysmal nocturnal hemoglobinuria patients treated or not by eculizumab: Focus on platelet extracellular vesicles",
abstract = "Thrombosis are severe complications of paroxysmal nocturnal hemoglobinuria (PNH), effectively reduced by eculizumab. Extracellular vesicles (EVs) may play a central role. The objective of this study was to assess the procoagulant activity of plasma isolated from PNH patients (treated or not by eculizumab) and to quantify their circulating EVs.We iteratively collected the platelet-free-plasma of 17 PNH patients and 16 matched healthy volunteers, quantified their circulating EVs by flow cytometry and evaluated their procoagulant activity by thrombin generation and STA-Procoag-procoagulant phospholipid (PPL) assays.A significant decrease of EVs from platelets (P = .024) and an increase of the STA-Procoag-PPL clotting time (P = .049) was observed after initiation of eculizumab and up to 11 weeks after. This reduction of prothrombotic biomarkers was not observed with the thrombin generation test due to a lack of sensitivity of this assay. Active hemolysis was observed in 90{\%} of patients and elevated D-dimers in 41{\%} of them. However, no significant difference was observed between patients and control subjects regarding the procoagulant activity, the EVs quantity, or the cellular origin. Lactate dehydrogenase (LDH) levels were lower in eculizumab-treated patients compared to nontreated patients (441 vs 2448 IU/L). D-dimers and LDH decreased after administration of eculizumab (mean decrease of 1307 ng/mL and 4159 IU/L, respectively).These observations suggest a decrease of the phospholipid-dependent procoagulant potential of EVs after eculizumab therapy in PNH patients. TRIAL REGISTRATION:: NUB: B039201214365.",
keywords = "Administration, Intravenous, Adult, Aged, Antibodies, Monoclonal, Humanized/administration & dosage, Case-Control Studies, Extracellular Vesicles/drug effects, Flow Cytometry, Hemoglobinuria, Paroxysmal/blood, Humans, Middle Aged, Prospective Studies, Thrombosis/etiology",
author = "B{\'e}rang{\`e}re Devalet and Adeline Wannez and Nicolas Bailly and Lutfiye Alpan and Damien Gheldof and Jonathan Douxfils and Beno{\^i}t Bihin and Bernard Chatelain and Jean-Michel Dogn{\'e} and Christian Chatelain and Fran{\cc}ois Mullier",
year = "2019",
month = "7",
day = "1",
doi = "10.1097/MD.0000000000016164",
language = "English",
volume = "98",
pages = "e16164",
journal = "Medicine (Mumbai): analytical reviews of general medicine, neurology, psychiatry, dermatology and pediatrics",
issn = "0025-7974",
publisher = "Wolters Kluwer",
number = "27",

}

TY - JOUR

T1 - Prospective and comparative study of paroxysmal nocturnal hemoglobinuria patients treated or not by eculizumab

T2 - Focus on platelet extracellular vesicles

AU - Devalet, Bérangère

AU - Wannez, Adeline

AU - Bailly, Nicolas

AU - Alpan, Lutfiye

AU - Gheldof, Damien

AU - Douxfils, Jonathan

AU - Bihin, Benoît

AU - Chatelain, Bernard

AU - Dogné, Jean-Michel

AU - Chatelain, Christian

AU - Mullier, François

PY - 2019/7/1

Y1 - 2019/7/1

N2 - Thrombosis are severe complications of paroxysmal nocturnal hemoglobinuria (PNH), effectively reduced by eculizumab. Extracellular vesicles (EVs) may play a central role. The objective of this study was to assess the procoagulant activity of plasma isolated from PNH patients (treated or not by eculizumab) and to quantify their circulating EVs.We iteratively collected the platelet-free-plasma of 17 PNH patients and 16 matched healthy volunteers, quantified their circulating EVs by flow cytometry and evaluated their procoagulant activity by thrombin generation and STA-Procoag-procoagulant phospholipid (PPL) assays.A significant decrease of EVs from platelets (P = .024) and an increase of the STA-Procoag-PPL clotting time (P = .049) was observed after initiation of eculizumab and up to 11 weeks after. This reduction of prothrombotic biomarkers was not observed with the thrombin generation test due to a lack of sensitivity of this assay. Active hemolysis was observed in 90% of patients and elevated D-dimers in 41% of them. However, no significant difference was observed between patients and control subjects regarding the procoagulant activity, the EVs quantity, or the cellular origin. Lactate dehydrogenase (LDH) levels were lower in eculizumab-treated patients compared to nontreated patients (441 vs 2448 IU/L). D-dimers and LDH decreased after administration of eculizumab (mean decrease of 1307 ng/mL and 4159 IU/L, respectively).These observations suggest a decrease of the phospholipid-dependent procoagulant potential of EVs after eculizumab therapy in PNH patients. TRIAL REGISTRATION:: NUB: B039201214365.

AB - Thrombosis are severe complications of paroxysmal nocturnal hemoglobinuria (PNH), effectively reduced by eculizumab. Extracellular vesicles (EVs) may play a central role. The objective of this study was to assess the procoagulant activity of plasma isolated from PNH patients (treated or not by eculizumab) and to quantify their circulating EVs.We iteratively collected the platelet-free-plasma of 17 PNH patients and 16 matched healthy volunteers, quantified their circulating EVs by flow cytometry and evaluated their procoagulant activity by thrombin generation and STA-Procoag-procoagulant phospholipid (PPL) assays.A significant decrease of EVs from platelets (P = .024) and an increase of the STA-Procoag-PPL clotting time (P = .049) was observed after initiation of eculizumab and up to 11 weeks after. This reduction of prothrombotic biomarkers was not observed with the thrombin generation test due to a lack of sensitivity of this assay. Active hemolysis was observed in 90% of patients and elevated D-dimers in 41% of them. However, no significant difference was observed between patients and control subjects regarding the procoagulant activity, the EVs quantity, or the cellular origin. Lactate dehydrogenase (LDH) levels were lower in eculizumab-treated patients compared to nontreated patients (441 vs 2448 IU/L). D-dimers and LDH decreased after administration of eculizumab (mean decrease of 1307 ng/mL and 4159 IU/L, respectively).These observations suggest a decrease of the phospholipid-dependent procoagulant potential of EVs after eculizumab therapy in PNH patients. TRIAL REGISTRATION:: NUB: B039201214365.

KW - Administration, Intravenous

KW - Adult

KW - Aged

KW - Antibodies, Monoclonal, Humanized/administration & dosage

KW - Case-Control Studies

KW - Extracellular Vesicles/drug effects

KW - Flow Cytometry

KW - Hemoglobinuria, Paroxysmal/blood

KW - Humans

KW - Middle Aged

KW - Prospective Studies

KW - Thrombosis/etiology

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U2 - 10.1097/MD.0000000000016164

DO - 10.1097/MD.0000000000016164

M3 - Article

C2 - 31277120

VL - 98

SP - e16164

JO - Medicine (Mumbai): analytical reviews of general medicine, neurology, psychiatry, dermatology and pediatrics

JF - Medicine (Mumbai): analytical reviews of general medicine, neurology, psychiatry, dermatology and pediatrics

SN - 0025-7974

IS - 27

ER -