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Thromb Haemost
DOI: 10.1055/a-2498-4849
Endothelium and Angiogenesis

Endothelial Damage in JAK2V617F Myeloproliferative Neoplasms with Splanchnic Vein Thrombosis

Blanca De Moner 
1   Hemostasis and Erythropathology Laboratory, Hematopathology, Pathology Department, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain
2   Josep Carreras Leukaemia Research Institute, University of Barcelona, Hospital Clínic de Barcelona, Barcelona, Spain
3   Barcelona Endothelium Team, Barcelona, Spain
,
Julia Martinez-Sanchez
1   Hemostasis and Erythropathology Laboratory, Hematopathology, Pathology Department, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain
3   Barcelona Endothelium Team, Barcelona, Spain
,
Marta Garrote
4   Hematopathology Section, Pathology Department, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, IDIBAPS, Spain
,
Alex Ramos
1   Hemostasis and Erythropathology Laboratory, Hematopathology, Pathology Department, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain
3   Barcelona Endothelium Team, Barcelona, Spain
,
Helena Ventosa-Capell
5   Medical Intensive Care Unit, Hospital Clínic de Barcelona, Barcelona, Spain
,
Ana Moreno-Castaño
1   Hemostasis and Erythropathology Laboratory, Hematopathology, Pathology Department, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain
3   Barcelona Endothelium Team, Barcelona, Spain
,
Meritxell Nomdedeu
6   Hemostasis and Hemotherapy Department, Institut del Càncer i Malalties de la Sang (ICAMS), Hospital Clínic de Barcelona, Barcelona, Spain
,
Asunción Ojeda
7   Liver Unit, Barcelona Hepatic Haemodynamic Laboratory, Hospital Clínic de Barcelona, IDIBAPS and CIBERehd, Barcelona, Spain
,
Gines Escolar
1   Hemostasis and Erythropathology Laboratory, Hematopathology, Pathology Department, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain
3   Barcelona Endothelium Team, Barcelona, Spain
,
Joan Carles Garcia-Pagan
7   Liver Unit, Barcelona Hepatic Haemodynamic Laboratory, Hospital Clínic de Barcelona, IDIBAPS and CIBERehd, Barcelona, Spain
,
Eduardo Arellano-Rodrigo 
6   Hemostasis and Hemotherapy Department, Institut del Càncer i Malalties de la Sang (ICAMS), Hospital Clínic de Barcelona, Barcelona, Spain
,
Enric Carreras
3   Barcelona Endothelium Team, Barcelona, Spain
8   Fundació Carreras contra la Leucèmia, Barcelona, Spain
,
Alberto Alvarez-Larran   *
9   Hematology Department, Institut del Càncer i Malalties de la Sang (ICAMS), Hospital Clínic de Barcelona, IDIBAPS, Barcelona, Spain
,
Maribel Díaz-Ricart   *
1   Hemostasis and Erythropathology Laboratory, Hematopathology, Pathology Department, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain
3   Barcelona Endothelium Team, Barcelona, Spain
› Author Affiliations Funding The study was partially supported by Instituto de Salud Carlos III (PI18/00205, PI19/00888) integrados en el Plan Nacional de I + D + I y cofinanciados por el ISCIII-Subdirección General de Evaluación y el Fondo Europeo de Desarrollo Regional (FEDER) “Una manera de hacer Europa” (Spain); Departament de Recerca i Universitats de la Generalitat de Catalunya (2021-SGR-01118); Contrato Clínico de investigación “Emili Letang-Josep Font”, Novartis Farmacéutica SA.
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Abstract

Background JAK2V617F-mutated myeloproliferative neoplasms (MPN) exhibit abnormal proliferation of bone marrow progenitors and increased risk of thrombosis, specifically in splanchnic veins (SVT). The contribution of the endothelium to the development of the prothrombotic phenotype was explored.

Material and Methods Plasma and serum samples from JAK2V617F MPN patients with (n=26) or without (n=7) thrombotic debut and different treatments, were obtained (n=33). Cultured endothelial cells (ECs) were exposed to serum samples from these patients and from healthy donors as controls. Changes in markers of inflammation (VCAM-1, ICAM-1), cell permeability (VE-cadherin), production of VWF, extracellular matrix (ECM) reactivity, and activation of intracellular signaling pathways related to stress, proliferation, inflammation (Akt, p44/42, IkBa), and JAK2/STAT3 pathway, were assessed by immunofluorescence, flow adhesion, SDS-PAGE and immunoblot. Additionally, circulating markers of endothelial activation and damage (VWF, sVCAM-1, sTNFRI, thrombomodulin, angiopoietin-2, a2-antiplasmin activity, PAI-1) were evaluated in Patients' plasma.

Results The in vitro studies showed that EC exposure to MPN thrombotic patients' sera resulted in increased VCAM-1 and ICAM-1, and reduced VE-cadherin expression (p<0.05) at the cell surface. Production and release of VWF to the ECM were higher (p<0.05), with increased platelet adhesion after perfusing whole blood, being more noticeable in response to sera from non-treated patients. Furthermore, intracellular activation of Akt, p44/42, IkBa and JAK2/STAT3 was observed. Moreover, plasma levels of VWF, TNF-R1, VCAM-1, thrombomodulin, and angiopoietin-2 were higher in JAK2V617F+ MPN patients with thrombosis.

Conclusion The present findings suggest that circulating factors in MPNs with SVT debut induce endothelial proinflammatory and prothrombotic phenotypes, which are modulated in vitro with MPN treatment.

Keywords

endothelial cells - myeloproliferative neoplasms - splanchnic vein thrombosis - JAK2V617F - inflammation

Authors' Contribution

BDM designed and performed the experiments, analyzed the results, designed the images, and wrote the manuscript. JM-S, MG, AR, and HV-C participated in the experiments and analysis of the results. MN, GE, JCG-P, and EC reviewed the results and manuscript. EA-R, AA-L, and MD-R designed, supervised, and reviewed the study, results, and manuscript.


* These authors contributed equally to this work.


Supplementary Material



Publication History

Received: 27 August 2024

Accepted: 07 December 2024

Article published online:
14 January 2025

© 2025. Thieme. All rights reserved.

Georg Thieme Verlag KG
Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany

 

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