Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic mice

Background: Elevated von Willebrand factor (VWF) levels correlate with higher risk of atherosclerosis-related arterial thrombosis (atherothrombosis). Silencing the VWF gene via small-interfering RNAs (siRNAs) could mitigate this risk. Previous studies successfully delivered siRNA to the endothelium...

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Main Authors: Yvonne K. Jongejan, Richard J. Dirven, Elisa Schrader Echeverri, Anke J.L. de Jong, Amanda C.M. Pronk, Sander Kooijman, Patrick C.N. Rensen, James E. Dahlman, Jeroen C.J. Eikenboom, Bart J.M. van Vlijmen
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Language:English
Published: Elsevier 2025-01-01
Series:Research and Practice in Thrombosis and Haemostasis
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Online Access:http://www.sciencedirect.com/science/article/pii/S2475037925000238
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author Yvonne K. Jongejan
Richard J. Dirven
Elisa Schrader Echeverri
Anke J.L. de Jong
Amanda C.M. Pronk
Sander Kooijman
Patrick C.N. Rensen
James E. Dahlman
Jeroen C.J. Eikenboom
Bart J.M. van Vlijmen
author_facet Yvonne K. Jongejan
Richard J. Dirven
Elisa Schrader Echeverri
Anke J.L. de Jong
Amanda C.M. Pronk
Sander Kooijman
Patrick C.N. Rensen
James E. Dahlman
Jeroen C.J. Eikenboom
Bart J.M. van Vlijmen
author_sort Yvonne K. Jongejan
collection DOAJ
description Background: Elevated von Willebrand factor (VWF) levels correlate with higher risk of atherosclerosis-related arterial thrombosis (atherothrombosis). Silencing the VWF gene via small-interfering RNAs (siRNAs) could mitigate this risk. Previous studies successfully delivered siRNA to the endothelium of healthy, wild-type (WT) mice using lipid nanoparticles (LNPs). Objectives: This study aimed to investigate whether the LNP-siRNA strategy could achieve endothelium-specific Vwf-silencing under diseased conditions of prolonged hypercholesterolemia and atherothrombosis-prone vasculature. Methods: Female transgenic mice expressing a variant of human APOE∗3 (ie, APOE∗3-Leiden) and human cholesteryl ester transfer protein (CETP), fed a cholesterol-enriched diet for 18 weeks, received an intravenous injection of LNP-encapsulated siRNA targeting Vwf (siVwf) or scrambled control siRNA at 1.5 mg siRNA/kg. For comparison, the same LNP-siRNAs were administered to young, chow-fed WT mice. Plasma VWF and Vwf mRNA levels were measured 96 hours after injection, with immunofluorescence analysis of lungs and heart aortic root to assess VWF protein expression. Results: APOE∗3-Leiden.CETP mice exhibited elevated plasma VWF levels compared with WT mice, alongside hypercholesterolemia and aortic atherosclerosis. siVwf administration led to over 85% reduction in plasma VWF in both strains, with a strong reduction in lung Vwf mRNA and VWF protein in the pulmonary endothelium. Similarly, siVwf treatment resulted in the virtual absence of VWF protein in the endothelial lining of the aortic root of both nondiseased (WT mice) and atherosclerotic (APOE∗3-Leiden.CETP mice) vessel walls. Conclusion: The LNP-siRNA targeting Vwf strongly reduced plasma and endothelial VWF in mice with hypercholesterolemia and advanced atherosclerosis, indicating feasibility to target endothelial VWF under proatherothrombotic conditions.
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spelling doaj-art-8f43e381eba447d782d3c31a552e14bd2025-08-20T03:04:38ZengElsevierResearch and Practice in Thrombosis and Haemostasis2475-03792025-01-019110269910.1016/j.rpth.2025.102699Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic miceYvonne K. Jongejan0Richard J. Dirven1Elisa Schrader Echeverri2Anke J.L. de Jong3Amanda C.M. Pronk4Sander Kooijman5Patrick C.N. Rensen6James E. Dahlman7Jeroen C.J. Eikenboom8Bart J.M. van Vlijmen9Division of Thrombosis and Hemostasis, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, NetherlandsDivision of Thrombosis and Hemostasis, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, NetherlandsWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, Georgia, USADivision of Thrombosis and Hemostasis, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, NetherlandsDivision of Endocrinology, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, NetherlandsDivision of Endocrinology, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, NetherlandsDivision of Endocrinology, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, NetherlandsWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, Georgia, USADivision of Thrombosis and Hemostasis, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, NetherlandsDivision of Thrombosis and Hemostasis, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, Netherlands; Correspondence Bart J. M. van Vlijmen, Division of Thrombosis and Haemostasis, Department of Internal Medicine, Leiden University Medical Center, Einthoven Laboratory for Vascular and Regenerative Medicine, P.O. Box 9600, 2300 RC Leiden, Netherlands.Background: Elevated von Willebrand factor (VWF) levels correlate with higher risk of atherosclerosis-related arterial thrombosis (atherothrombosis). Silencing the VWF gene via small-interfering RNAs (siRNAs) could mitigate this risk. Previous studies successfully delivered siRNA to the endothelium of healthy, wild-type (WT) mice using lipid nanoparticles (LNPs). Objectives: This study aimed to investigate whether the LNP-siRNA strategy could achieve endothelium-specific Vwf-silencing under diseased conditions of prolonged hypercholesterolemia and atherothrombosis-prone vasculature. Methods: Female transgenic mice expressing a variant of human APOE∗3 (ie, APOE∗3-Leiden) and human cholesteryl ester transfer protein (CETP), fed a cholesterol-enriched diet for 18 weeks, received an intravenous injection of LNP-encapsulated siRNA targeting Vwf (siVwf) or scrambled control siRNA at 1.5 mg siRNA/kg. For comparison, the same LNP-siRNAs were administered to young, chow-fed WT mice. Plasma VWF and Vwf mRNA levels were measured 96 hours after injection, with immunofluorescence analysis of lungs and heart aortic root to assess VWF protein expression. Results: APOE∗3-Leiden.CETP mice exhibited elevated plasma VWF levels compared with WT mice, alongside hypercholesterolemia and aortic atherosclerosis. siVwf administration led to over 85% reduction in plasma VWF in both strains, with a strong reduction in lung Vwf mRNA and VWF protein in the pulmonary endothelium. Similarly, siVwf treatment resulted in the virtual absence of VWF protein in the endothelial lining of the aortic root of both nondiseased (WT mice) and atherosclerotic (APOE∗3-Leiden.CETP mice) vessel walls. Conclusion: The LNP-siRNA targeting Vwf strongly reduced plasma and endothelial VWF in mice with hypercholesterolemia and advanced atherosclerosis, indicating feasibility to target endothelial VWF under proatherothrombotic conditions.http://www.sciencedirect.com/science/article/pii/S2475037925000238atherothrombosishypercholesterolemiamouse modelsiRNAvon Willebrand factor
spellingShingle Yvonne K. Jongejan
Richard J. Dirven
Elisa Schrader Echeverri
Anke J.L. de Jong
Amanda C.M. Pronk
Sander Kooijman
Patrick C.N. Rensen
James E. Dahlman
Jeroen C.J. Eikenboom
Bart J.M. van Vlijmen
Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic mice
Research and Practice in Thrombosis and Haemostasis
atherothrombosis
hypercholesterolemia
mouse model
siRNA
von Willebrand factor
title Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic mice
title_full Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic mice
title_fullStr Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic mice
title_full_unstemmed Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic mice
title_short Silencing of the von Willebrand factor gene in proatherothrombotic APOE∗3-Leiden.CETP transgenic mice
title_sort silencing of the von willebrand factor gene in proatherothrombotic apoe∗3 leiden cetp transgenic mice
topic atherothrombosis
hypercholesterolemia
mouse model
siRNA
von Willebrand factor
url http://www.sciencedirect.com/science/article/pii/S2475037925000238
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