SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis

Julia Tilburg; Danielle M. Coenen; Gaia Zirka; Sophie Dolleman; Annemarie M. van Oeveren-Rietdijk; Mieke F. A. Karel; Hetty C. de Boer; Judith M. E. M. Cosemans; Henri H. Versteeg; Pierre E. Morange; Bart J. M. van Vlijmen; Chrissta X. Maracle; Grace M. Thomas

doi:10.1111/jth.14835

SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis

Julia Tilburg, Danielle M. Coenen, Gaia Zirka, Sophie Dolleman, Annemarie M. van Oeveren-Rietdijk, Mieke F. A. Karel, Hetty C. de Boer, Judith M. E. M. Cosemans, Henri H. Versteeg, Pierre E. Morange, Bart J. M. van Vlijmen^*, Chrissta X. Maracle, Grace M. Thomas

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Background Genome wide association studies (GWAS) identified SLC44A2 as a novel susceptibility gene for venous thrombosis (VT) and previous work established that SLC44A2 contributed to clot formation upon vascular injury.

Objective To further investigate the role of SLC44A2 in VT by utilizing SLC44A2 deficient mice (Slc44a2(-)(/)(-)) in two representative disease models.

Methods Mice were included in a hypercoagulability model driven by siRNA-mediated hepatic gene silencing of anticoagulants Serpinc1 (antithrombin) and Proc (protein C) and a flow restriction (stenosis) model induced by partial ligation of the inferior vena cava.

Results In the hypercoagulability model, no effect in onset was observed in Slc44a2(-)(/)(-) animals; however, a drop in plasma fibrinogen and von Willebrand factor coinciding with an increase in blood neutrophils was recorded. In the neutrophil dependent stenosis model after 48 hours, Slc44a2(-)(/)(-) mice had significantly smaller thrombi both in length and weight with less platelet accumulation as a percentage of the total thrombus area. During the initiation of thrombosis at 6 hours post-stenosis, Slc44a2(-)(/)(-) mice also had smaller thrombi both in length and weight, with circulating platelets remaining elevated in Slc44a2(-)(/)(-) animals. Platelet activation and aggregation under both static- and venous and arterial shear conditions were normal for blood from Slc44a2(-)(/)(-) mice.

Conclusions These studies corroborate the original GWAS findings and establish a contributing role for SLC44A2 during the initiation of VT, with indications that this may be related to platelet-neutrophil interaction. The precise mechanism however remains elusive and warrants further investigation.

Original language	English
Pages (from-to)	1714-1727
Number of pages	14
Journal	Journal of Thrombosis and Haemostasis
Volume	18
Issue number	7
DOIs	https://doi.org/10.1111/jth.14835
Publication status	Published - Jul 2020

Keywords

animal models of human disease
basic science research
thrombosis
vascular biology
vascular diseases
VON-WILLEBRAND-FACTOR
DEEP-VEIN THROMBOSIS
TRANSPORTER-LIKE PROTEIN-2
FACTOR-BINDING
NEUTROPHILS
GLYCOPROTEIN
INDIVIDUALS
INITIATE
LOCI
LUNG

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Tilburg, J., Coenen, D. M., Zirka, G., Dolleman, S., van Oeveren-Rietdijk, A. M., Karel, M. F. A., de Boer, H. C., Cosemans, J. M. E. M., Versteeg, H. H., Morange, P. E., van Vlijmen, B. J. M., Maracle, C. X., & Thomas, G. M. (2020). SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis. Journal of Thrombosis and Haemostasis, 18(7), 1714-1727. https://doi.org/10.1111/jth.14835

@article{65410a1a458e4e26b74c59a5714ec1f9,

title = "SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis",

abstract = "Background Genome wide association studies (GWAS) identified SLC44A2 as a novel susceptibility gene for venous thrombosis (VT) and previous work established that SLC44A2 contributed to clot formation upon vascular injury.Objective To further investigate the role of SLC44A2 in VT by utilizing SLC44A2 deficient mice (Slc44a2(-)(/)(-)) in two representative disease models.Methods Mice were included in a hypercoagulability model driven by siRNA-mediated hepatic gene silencing of anticoagulants Serpinc1 (antithrombin) and Proc (protein C) and a flow restriction (stenosis) model induced by partial ligation of the inferior vena cava.Results In the hypercoagulability model, no effect in onset was observed in Slc44a2(-)(/)(-) animals; however, a drop in plasma fibrinogen and von Willebrand factor coinciding with an increase in blood neutrophils was recorded. In the neutrophil dependent stenosis model after 48 hours, Slc44a2(-)(/)(-) mice had significantly smaller thrombi both in length and weight with less platelet accumulation as a percentage of the total thrombus area. During the initiation of thrombosis at 6 hours post-stenosis, Slc44a2(-)(/)(-) mice also had smaller thrombi both in length and weight, with circulating platelets remaining elevated in Slc44a2(-)(/)(-) animals. Platelet activation and aggregation under both static- and venous and arterial shear conditions were normal for blood from Slc44a2(-)(/)(-) mice.Conclusions These studies corroborate the original GWAS findings and establish a contributing role for SLC44A2 during the initiation of VT, with indications that this may be related to platelet-neutrophil interaction. The precise mechanism however remains elusive and warrants further investigation.",

keywords = "animal models of human disease, basic science research, thrombosis, vascular biology, vascular diseases, VON-WILLEBRAND-FACTOR, DEEP-VEIN THROMBOSIS, TRANSPORTER-LIKE PROTEIN-2, FACTOR-BINDING, NEUTROPHILS, GLYCOPROTEIN, INDIVIDUALS, INITIATE, LOCI, LUNG",

author = "Julia Tilburg and Coenen, {Danielle M.} and Gaia Zirka and Sophie Dolleman and {van Oeveren-Rietdijk}, {Annemarie M.} and Karel, {Mieke F. A.} and {de Boer}, {Hetty C.} and Cosemans, {Judith M. E. M.} and Versteeg, {Henri H.} and Morange, {Pierre E.} and {van Vlijmen}, {Bart J. M.} and Maracle, {Chrissta X.} and Thomas, {Grace M.}",

note = "Funding Information: This work was supported by Trombosestichting Nederland (#2015-4), the Landsteiner Foundation for Blood Transfusion Research (#1503), the Dutch Heart Foundation (2015T79), the Netherlands Organization for Scientific Research (NWO Vidi 91716421), a European Molecular Biology Organization Short-Term Fellowship (EMBO #7468), a European Society of Cardiology First Contact Initiative grant, and by a French ANR grant (ANR-17-CE14-0003-01). The authors thank Dr Pavan K. Kommareddi, Thankam S. Nair, and Dr Thomas E. Carey (Kresge Hearing Research Institute, Department of Otolaryngology/Head and Neck Surgery, University of Michigan, Ann Arbor, MI, USA) for the floxed Slc44a2 mouse embryos used to generate the mice on a C57BL/6J background and Dr Charles Esmon (University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA) for providing the 59D8 fibrin antibody. Also, we thank Lieke van den Heijkant for assisting with the hypercoagulability model. Funding Information: This work was supported by Trombosestichting Nederland (#2015‐4), the Landsteiner Foundation for Blood Transfusion Research (#1503), the Dutch Heart Foundation (2015T79), the Netherlands Organization for Scientific Research (NWO Vidi 91716421), a European Molecular Biology Organization Short‐Term Fellowship (EMBO #7468), a European Society of Cardiology First Contact Initiative grant, and by a French ANR grant (ANR‐17‐CE14‐0003‐01). Publisher Copyright: {\textcopyright} 2020 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis",

year = "2020",

month = jul,

doi = "10.1111/jth.14835",

language = "English",

volume = "18",

pages = "1714--1727",

journal = "Journal of Thrombosis and Haemostasis",

issn = "1538-7933",

publisher = "Elsevier B.V.",

number = "7",

}

Tilburg, J, Coenen, DM, Zirka, G, Dolleman, S, van Oeveren-Rietdijk, AM, Karel, MFA, de Boer, HC, Cosemans, JMEM, Versteeg, HH, Morange, PE, van Vlijmen, BJM, Maracle, CX & Thomas, GM 2020, 'SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis', Journal of Thrombosis and Haemostasis, vol. 18, no. 7, pp. 1714-1727. https://doi.org/10.1111/jth.14835

TY - JOUR

T1 - SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis

AU - Tilburg, Julia

AU - Coenen, Danielle M.

AU - Zirka, Gaia

AU - Dolleman, Sophie

AU - van Oeveren-Rietdijk, Annemarie M.

AU - Karel, Mieke F. A.

AU - de Boer, Hetty C.

AU - Cosemans, Judith M. E. M.

AU - Versteeg, Henri H.

AU - Morange, Pierre E.

AU - van Vlijmen, Bart J. M.

AU - Maracle, Chrissta X.

AU - Thomas, Grace M.

N1 - Funding Information: This work was supported by Trombosestichting Nederland (#2015-4), the Landsteiner Foundation for Blood Transfusion Research (#1503), the Dutch Heart Foundation (2015T79), the Netherlands Organization for Scientific Research (NWO Vidi 91716421), a European Molecular Biology Organization Short-Term Fellowship (EMBO #7468), a European Society of Cardiology First Contact Initiative grant, and by a French ANR grant (ANR-17-CE14-0003-01). The authors thank Dr Pavan K. Kommareddi, Thankam S. Nair, and Dr Thomas E. Carey (Kresge Hearing Research Institute, Department of Otolaryngology/Head and Neck Surgery, University of Michigan, Ann Arbor, MI, USA) for the floxed Slc44a2 mouse embryos used to generate the mice on a C57BL/6J background and Dr Charles Esmon (University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA) for providing the 59D8 fibrin antibody. Also, we thank Lieke van den Heijkant for assisting with the hypercoagulability model. Funding Information: This work was supported by Trombosestichting Nederland (#2015‐4), the Landsteiner Foundation for Blood Transfusion Research (#1503), the Dutch Heart Foundation (2015T79), the Netherlands Organization for Scientific Research (NWO Vidi 91716421), a European Molecular Biology Organization Short‐Term Fellowship (EMBO #7468), a European Society of Cardiology First Contact Initiative grant, and by a French ANR grant (ANR‐17‐CE14‐0003‐01). Publisher Copyright: © 2020 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis

PY - 2020/7

Y1 - 2020/7

N2 - Background Genome wide association studies (GWAS) identified SLC44A2 as a novel susceptibility gene for venous thrombosis (VT) and previous work established that SLC44A2 contributed to clot formation upon vascular injury.Objective To further investigate the role of SLC44A2 in VT by utilizing SLC44A2 deficient mice (Slc44a2(-)(/)(-)) in two representative disease models.Methods Mice were included in a hypercoagulability model driven by siRNA-mediated hepatic gene silencing of anticoagulants Serpinc1 (antithrombin) and Proc (protein C) and a flow restriction (stenosis) model induced by partial ligation of the inferior vena cava.Results In the hypercoagulability model, no effect in onset was observed in Slc44a2(-)(/)(-) animals; however, a drop in plasma fibrinogen and von Willebrand factor coinciding with an increase in blood neutrophils was recorded. In the neutrophil dependent stenosis model after 48 hours, Slc44a2(-)(/)(-) mice had significantly smaller thrombi both in length and weight with less platelet accumulation as a percentage of the total thrombus area. During the initiation of thrombosis at 6 hours post-stenosis, Slc44a2(-)(/)(-) mice also had smaller thrombi both in length and weight, with circulating platelets remaining elevated in Slc44a2(-)(/)(-) animals. Platelet activation and aggregation under both static- and venous and arterial shear conditions were normal for blood from Slc44a2(-)(/)(-) mice.Conclusions These studies corroborate the original GWAS findings and establish a contributing role for SLC44A2 during the initiation of VT, with indications that this may be related to platelet-neutrophil interaction. The precise mechanism however remains elusive and warrants further investigation.

AB - Background Genome wide association studies (GWAS) identified SLC44A2 as a novel susceptibility gene for venous thrombosis (VT) and previous work established that SLC44A2 contributed to clot formation upon vascular injury.Objective To further investigate the role of SLC44A2 in VT by utilizing SLC44A2 deficient mice (Slc44a2(-)(/)(-)) in two representative disease models.Methods Mice were included in a hypercoagulability model driven by siRNA-mediated hepatic gene silencing of anticoagulants Serpinc1 (antithrombin) and Proc (protein C) and a flow restriction (stenosis) model induced by partial ligation of the inferior vena cava.Results In the hypercoagulability model, no effect in onset was observed in Slc44a2(-)(/)(-) animals; however, a drop in plasma fibrinogen and von Willebrand factor coinciding with an increase in blood neutrophils was recorded. In the neutrophil dependent stenosis model after 48 hours, Slc44a2(-)(/)(-) mice had significantly smaller thrombi both in length and weight with less platelet accumulation as a percentage of the total thrombus area. During the initiation of thrombosis at 6 hours post-stenosis, Slc44a2(-)(/)(-) mice also had smaller thrombi both in length and weight, with circulating platelets remaining elevated in Slc44a2(-)(/)(-) animals. Platelet activation and aggregation under both static- and venous and arterial shear conditions were normal for blood from Slc44a2(-)(/)(-) mice.Conclusions These studies corroborate the original GWAS findings and establish a contributing role for SLC44A2 during the initiation of VT, with indications that this may be related to platelet-neutrophil interaction. The precise mechanism however remains elusive and warrants further investigation.

KW - animal models of human disease

KW - basic science research

KW - thrombosis

KW - vascular biology

KW - vascular diseases

KW - VON-WILLEBRAND-FACTOR

KW - DEEP-VEIN THROMBOSIS

KW - TRANSPORTER-LIKE PROTEIN-2

KW - FACTOR-BINDING

KW - NEUTROPHILS

KW - GLYCOPROTEIN

KW - INDIVIDUALS

KW - INITIATE

KW - LOCI

KW - LUNG

U2 - 10.1111/jth.14835

DO - 10.1111/jth.14835

M3 - Article

C2 - 32297475

SN - 1538-7933

VL - 18

SP - 1714

EP - 1727

JO - Journal of Thrombosis and Haemostasis

JF - Journal of Thrombosis and Haemostasis

IS - 7

ER -

SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis

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