Factor VIIa releases phosphatidylserine-enriched extracellular vesicles from endothelial cells by activating acid sphingomyelinase.


Journal

Journal of thrombosis and haemostasis : JTH
ISSN: 1538-7836
Titre abrégé: J Thromb Haemost
Pays: England
ID NLM: 101170508

Informations de publication

Date de publication:
12 2023
Historique:
received: 16 03 2023
revised: 05 08 2023
accepted: 23 08 2023
medline: 27 11 2023
pubmed: 25 10 2023
entrez: 24 10 2023
Statut: ppublish

Résumé

Our recent studies showed that activated factor (F) VII (FVIIa) releases extracellular vesicles (EVs) from the endothelium. FVIIa-released EVs were found to be enriched with phosphatidylserine (PS) and contribute to the hemostatic effect of FVIIa in thrombocytopenia and hemophilia. To investigate mechanisms by which FVIIa induces EV biogenesis and enriches EVs with PS. FVIIa activation of acid sphingomyelinase (aSMase) was evaluated by its translocation to the cell surface. The role of aSMase in the biogenesis of FVIIa-induced EVs and their enrichment with PS was investigated using specific siRNAs and inhibitors of aSMase and its downstream metabolites. Wild-type and aSMase FVIIa activation of aSMase is responsible for both the externalization of PS and the release of EVs in endothelial cells. FVIIa-induced aSMase activation led to ceramide generation and de novo expression of transmembrane protein 16F. Inhibitors of ceramidases, sphingosine kinase, or sphingosine-1-phosphate receptor modulator blocked FVIIa-induced expression of transmembrane protein 16F and PS externalization without interfering with FVIIa release of EVs. In vivo, FVIIa release of EVs was markedly impaired in aSMase Our study identifies a novel mechanism by which FVIIa induces PS externalization and releases PS-enriched EVs.

Sections du résumé

BACKGROUND
Our recent studies showed that activated factor (F) VII (FVIIa) releases extracellular vesicles (EVs) from the endothelium. FVIIa-released EVs were found to be enriched with phosphatidylserine (PS) and contribute to the hemostatic effect of FVIIa in thrombocytopenia and hemophilia.
OBJECTIVE
To investigate mechanisms by which FVIIa induces EV biogenesis and enriches EVs with PS.
METHODS
FVIIa activation of acid sphingomyelinase (aSMase) was evaluated by its translocation to the cell surface. The role of aSMase in the biogenesis of FVIIa-induced EVs and their enrichment with PS was investigated using specific siRNAs and inhibitors of aSMase and its downstream metabolites. Wild-type and aSMase
RESULTS
FVIIa activation of aSMase is responsible for both the externalization of PS and the release of EVs in endothelial cells. FVIIa-induced aSMase activation led to ceramide generation and de novo expression of transmembrane protein 16F. Inhibitors of ceramidases, sphingosine kinase, or sphingosine-1-phosphate receptor modulator blocked FVIIa-induced expression of transmembrane protein 16F and PS externalization without interfering with FVIIa release of EVs. In vivo, FVIIa release of EVs was markedly impaired in aSMase
CONCLUSION
Our study identifies a novel mechanism by which FVIIa induces PS externalization and releases PS-enriched EVs.

Identifiants

pubmed: 37875382
pii: S1538-7836(23)00654-2
doi: 10.1016/j.jtha.2023.08.025
pii:
doi:

Substances chimiques

Factor VIIa EC 3.4.21.21
Hemostatics 0
Phosphatidylserines 0
Sphingomyelin Phosphodiesterase EC 3.1.4.12
ASMase, mouse EC 3.1.4.12

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

3414-3431

Subventions

Organisme : NHLBI NIH HHS
ID : R01 HL124055
Pays : United States

Informations de copyright

Copyright © 2023 International Society on Thrombosis and Haemostasis. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of competing interests There are no competing interests to disclose.

Auteurs

Kaushik Das (K)

Department of Cellular and Molecular Biology, UT Tyler School of Medicine, The University of Texas Health Science Center at Tyler, Tyler, TX, USA. Electronic address: Kaushik.Das@uthct.edu.

Shiva Keshava (S)

Department of Cellular and Molecular Biology, UT Tyler School of Medicine, The University of Texas Health Science Center at Tyler, Tyler, TX, USA.

Tanmoy Mukherjee (T)

Department of Cellular and Molecular Biology, UT Tyler School of Medicine, The University of Texas Health Science Center at Tyler, Tyler, TX, USA.

Jue Wang (J)

Department of Cellular and Molecular Biology, UT Tyler School of Medicine, The University of Texas Health Science Center at Tyler, Tyler, TX, USA.

Jhansi Magisetty (J)

Department of Cellular and Molecular Biology, UT Tyler School of Medicine, The University of Texas Health Science Center at Tyler, Tyler, TX, USA.

Richard Kolesnick (R)

Memorial Sloan Kettering Cancer Center, New York, NY, USA.

Usha R Pendurthi (UR)

Department of Cellular and Molecular Biology, UT Tyler School of Medicine, The University of Texas Health Science Center at Tyler, Tyler, TX, USA.

L Vijaya Mohan Rao (LVM)

Department of Cellular and Molecular Biology, UT Tyler School of Medicine, The University of Texas Health Science Center at Tyler, Tyler, TX, USA. Electronic address: Vijay.Rao@uthct.edu.

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Classifications MeSH