Selective Estrogen Receptor Modulator Idoxifene Inhibits Smooth Muscle Cell Proliferation, Enhances Reendothelialization, and Inhibits Neointimal Formation In Vivo After Vascular Injury

Circulation ◽  
2000 ◽  
Vol 102 (suppl_3) ◽  
Author(s):  
Tian-Li Yue ◽  
Lynne Vickery-Clark ◽  
Calvert S. Louden ◽  
Juan-Li Gu ◽  
Xin L. Ma ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Smooth Muscle ◽  
Vascular Injury ◽  
Von Willebrand Factor ◽  
Carotid Arteries ◽  
Treated Group ◽  
Vehicle Group ◽  
Receptor Modulator ◽  
Von Willebrand ◽  
Willebrand Factor

Background —Idoxifene (ID) is a tissue-selective estrogen receptor modulator (SERM). The pharmacological profile of ID in animal studies suggests that it behaves like an estrogen receptor (ER) agonist in bone and lipid metabolism while having negligible ER activity on the reproductive system. It is unknown whether ID retains the vascular protective effects of estrogen. Methods and Results —In cultured vascular smooth muscle cells (VSMCs), ID inhibited platelet-derived growth factor–induced DNA synthesis and mitogenesis with IC 50 values of 20.4 and 27.5 nmol/L, respectively. Treatment with ID resulted in S-phase cell cycle arrest in serum-stimulated VSMCs. ID 1 to 100 nmol/L significantly protected endothelial cells from tumor necrosis factor-α (TNF-α)–induced apoptosis in vitro. Virgin Sprague-Dawley rats ovariectomized 1 week before the study were treated with ID (1 mg · kg − 1 · d − 1 ) or vehicle by gavage for 3 days before balloon denudation in carotid artery. The SMC proliferation in injured vessels was determined by immunostaining for proliferating cell nuclear antigen (PCNA). The number of PCNA-positive SMCs was reduced by 69%, 82%, and 86% in the media at days 1, 3 and 7, respectively, and by 78% in the neointima at day 7 after injury in ID- versus vehicle-treated group ( P <0.01). ID significantly enhanced reendothelialization in the injured carotid arteries as determined by Evans blue stain and immunohistochemical analysis for von Willebrand factor. In the former assay, the reendothelialized area in injured vessels was 43% in ID-treated group versus 24% in the vehicle group ( P <0.05); in the latter assay, the numbers of von Willebrand factor–positive cells per cross section increased from 24.8 (vehicle) to 60.5 (ID) ( P <0.01) at day 14 after injury. In addition, the production of nitric oxide from excised carotid arteries was significantly higher in ID-treated than the vehicle group (8.5 versus 2.7 nmol/g, P <0.01). Finally, ID treatment reduced neointimal area and the ratio of intima to media by 45% and 40%, respectively ( P <0.01), at day 14 after balloon angioplasty. Conclusions —The results indicate that ID beneficially modulates the balloon denudation–induced vascular injury response. Inhibition of VSMC proliferation and acceleration of endothelial recovery likely mediate this protective effect of ID.

scholarly journals von Willebrand Factor Permeates Small Vessels in CADASIL and Inhibits Smooth Muscle Gene Expression

2011 ◽  
Vol 3 (1) ◽  
pp. 138-145 ◽  
Author(s):  
Xiaojie Zhang ◽  
He Meng ◽  
Mila Blaivas ◽  
Elisabeth J. Rushing ◽  
Brian E. Moore ◽  
...  
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Von Willebrand Factor ◽  
Small Vessels ◽  
Muscle Gene Expression ◽  
Muscle Gene ◽  
Von Willebrand ◽  
Willebrand Factor

scholarly journals The VWF/LRP4/αVβ3-axis represents a novel pathway regulating proliferation of human vascular smooth muscle cells

2021 ◽  
Author(s):  
Jérémy Lagrange ◽  
Morel E Worou ◽  
Jean-Baptiste Michel ◽  
Alexandre Raoul ◽  
Mélusine Didelot ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Von Willebrand Factor ◽  
Vessel Wall ◽  
Integrin Αvβ3 ◽  
Vsmc Proliferation ◽  
Atherosclerotic Lesions ◽  
Von Willebrand ◽  
A2 Domain ◽  
Willebrand Factor

Abstract Aims Von Willebrand factor (VWF) is a plasma glycoprotein involved in primary hemostasis, while also having additional roles beyond hemostasis namely in cancer, inflammation, angiogenesis and potentially in vascular smooth muscle cell (VSMC) proliferation. Here, we addressed how VWF modulates VSMC proliferation and investigated the underlying molecular pathways and the in vivo pathophysiological relevance. Methods and results VWF induced proliferation of human aortic VSMCs and also promoted VSMC migration. Treatment of cells with a siRNA against αv integrin or the RGT-peptide blocking αvβ3 signaling abolished proliferation. However, VWF did not bind to αvβ3 on VSMCs through its RGD-motif. Rather, we identified the VWF A2 domain as the region mediating binding to the cells. We hypothesized the involvement of a member of the LDL-related receptor protein (LRP) family due to their known ability to act as co-receptors. Using the universal LRP-inhibitor receptor-associated protein, we confirmed LRP-mediated VSMC proliferation. siRNA experiments and confocal fluorescence microscopy identified LRP4 as the VWF-counterreceptor on VSMCs. Also co-localization between αvβ3 and LRP4 was observed via proximity ligation analysis and immuno-precipitation experiments. The pathophysiological relevance of our data was supported by VWF-deficient mice having significant reduced, if any, hyperplasia in carotid artery ligation and artery femoral denudation models. In wild-type mice, infiltration of VWF in intimal regions enriched in proliferating VSMCs was found. Interestingly, also analysis of human atherosclerotic lesions showed abundant VWF accumulation in VSMC-proliferating rich intimal areas. Conclusions VWF mediates VSMC proliferation through a mechanism involving A2 domain binding to the LRP4 receptor and integrin αvβ3 signaling. Our findings provide new insights into the mechanisms that drive physiological repair and pathological hyperplasia of the arterial vessel wall. In addition, the VWF/LRP4-axis may represent a novel therapeutic target to modulate VSMC proliferation. Translational perspective The mechanisms that drive physiological repair and pathological hyperplasia of the arterial vessel wall are complex and only partially understood. Specifically, the role of subendothelial-matrix proteins remains unclear. Here, we show that the hemostatic protein von Willebrand factor (VWF) accumulates in the vascular wall of atherosclerotic lesions and localizes to areas of vascular smooth muscle cell (VSMC) proliferation. VWF was found to use its A2-domain for binding to the VSMC-receptor LRP4, which in turn triggered outside-in signaling via integrin αVβ3, thereby inducing VSMC proliferation. Interfering with A2-domain/LRP4 interactions might offer innovative and additional therapeutical approaches to limit pathological hyperplasia.

Abstract 187: Reversible Aptamer Inhibition of Von Willebrand Factor is a Potent Thrombolytic and Ameliorates Stroke Burden Following Vascular Injury Compared to Recombinant Tissue Plasminogen Activator

Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
David L Dornbos ◽  
Hallie Harris ◽  
Surya Gnyawali ◽  
Debra G Wheeler ◽  
Allyson Huttinger ◽  
...  
Keyword(s):  
Carotid Artery ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Vascular Injury ◽  
Von Willebrand Factor ◽  
Recombinant Tissue Plasminogen Activator ◽  
Tissue Plasminogen ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Carotid Flow

Introduction: While recombinant tissue plasminogen activator (rTPA) is the mainstay of ischemic stroke treatment, few patients are eligible for treatment, and recanalization is only seen in 25-50%. Von Willebrand Factor (VWF) inhibition may play a role in thrombolysis. Hypothesis: VWF inhibition with an RNA aptamer lyses arterial thrombus and decreases ischemic injury. Furthermore, aptamer reversal with an antidote oligonucleotide ameliorates intracranial hemorrhage (ICH). Methods: Adult wild-type (C57BL/6J) mice were anesthetized, and the right carotid artery was exposed. Baseline carotid flow was obtained using a Doppler flow probe, and thrombotic occlusion was induced with a ferric chloride patch. After clot stabilization, mice were administered vehicle (platelet binding buffer, n=11), no infusion (n=8), rTPA (n=5) or VWF aptamer (n=5). Carotid flow was monitored for an additional 100 minutes. In a second cohort of mice, a 6-0 nylon suture was advanced within the carotid artery to generate vascular injury and ICH. Mice were given vehicle (n=16), rTPA (n=11), VWF aptamer (n=9) or aptamer/antidote (n=8). An MRI was obtained after 90 minutes to assess stroke and ICH volumes. Results: VWF aptamer successfully restored carotid blood flow 45 minutes following carotid occlusion (Figure 1) compared to controls (p<0.01*) and rTPA (p<0.05 + ). Stroke volume was significantly decreased in mice treated with VWF aptamer (23.03 ± 6.81 mm 3 ) and aptamer/antidote (12.48 ± 5.68 mm 3 ) compared to vehicle (45.25 ± 4.14 mm 3 , p<0.01). ICH volumes in mice treated with rTPA (2.64 ± 0.84 mm 3 ) were trending higher than vehicle (1.51 ± 0.17 mm 3 ), VWF aptamer (1.92 ± 0.22 mm 3 ) or aptamer/antidote (1.31 ± 0.35 mm 3 ). Conclusions: Aptamer inhibition of VWF is a potent thrombolytic agent with greater efficacy compared to rTPA. VWF inhibition appears safe with a trend toward lower ICH volumes in animals treated with aptamer and aptamer/antidote compared to rTPA.

Two Repressor Elements Inhibit Expression of the von Willebrand Factor Gene Promoter In Vitro

1998 ◽  
Vol 80 (09) ◽  
pp. 488-494 ◽  
Author(s):  
Ali Ardekani ◽  
Joel Greenberger ◽  
Nadia Jahroudi
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Von Willebrand Factor ◽  
Specific Activity ◽  
Regulatory Element ◽  
Gene Promoter ◽  
Muscle Cells ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryA fragment of the human von Willebrand factor (VWF) gene promoter corresponding to sequences –487 to +247 bp functions as an endothelial specific promoter in cell culture. We have previously reported that a GATA transcription factor functions as an activator and an NF1 like protein functions as a repressor of this promoter fragment. We have now identified a second negative regulatory element in the VWF promoter that interacts with nuclear factor(s) (designated R) in both bovine aortic endothelial and smooth muscle cells. Inhibition of either the NF1 or the R repressor alone is not sufficient to activate the VWF promoter in smooth muscle cells. The present studies reveal that simultaneous inhibition of both repressors activates the VWF promoter in smooth muscle cells. The data support a model of selective derepression to explain the endothelial cell specific activity of the –487 to +247 fragment of the VWF promoter in vitro.

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