Disturbed flow in radial-cephalic arteriovenous fistulae for haemodialysis: low and oscillating shear stress locates the sites of stenosis

Rationale: In diabetic animals as well as high glucose cell culture conditions, endothelial nitric oxide synthase (eNOS) is heavily O-GlcNAcylated, which inhibits its phosphorylation and nitric oxide (NO) production. It is unknown, however, whether varied blood flow conditions, which affect eNOS phosphorylation, modulate eNOS activity via O-GlcNAcylation-dependent mechanisms. Objective: The goal of this study was to test if steady laminar flow, but not oscillating disturbed flow, decreases eNOS O-GlcNAcylation, thereby elevating eNOS phosphorylation and NO production. Methods and Results: Human umbilical vein endothelial cells (HUVEC) were exposed to either laminar flow (20 dynes/cm2 shear stress) or oscillating disturbed flow (4{plus minus}6 dynes/cm2 shear stress) for 24 hours in a cone-and-plate device. eNOS O-GlcNAcylation was almost completely abolished in cells exposed to steady laminar but not oscillating disturbed flow. Interestingly, there was no change in protein level or activity of key O-GlcNAcylation enzymes (OGT, OGA, or GFAT). Instead, metabolomics data suggest that steady laminar flow decreases glycolysis and hexosamine biosynthetic pathway (HBP) activity, thereby reducing UDP-GlcNAc pool size and consequent O-GlcNAcylation. Inhibition of glycolysis via 2-deoxy-2-glucose (2-DG) in cells exposed to disturbed flow efficiently decreased eNOS O-GlcNAcylation, thereby increasing eNOS phosphorylation and NO production. Finally, we detected significantly higher O-GlcNAcylated proteins in endothelium of the inner aortic arch in mice, suggesting that disturbed flow increases protein O-GlcNAcylation in vivo. Conclusions: Our data demonstrate that steady laminar but not oscillating disturbed flow decreases eNOS O-GlcNAcylation by limiting glycolysis and UDP-GlcNAc substrate availability, thus enhancing eNOS phosphorylation and NO production. This research shows for the first time that O-GlcNAcylation is regulated by mechanical stimuli, relates flow-induced glycolytic reductions to macrovascular disease, and highlights targeting HBP metabolic enzymes in endothelial cells as a novel therapeutic strategy to restore eNOS activity and prevent EC dysfunction in cardiovascular disease.

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Abstract 363: Lim Domain Only 4 Is a Shear-Sensitive Protein, Playing a Critical Role in Endothelial Inflammation and Atherosclerosis

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.32.suppl_1.a363 ◽

2012 ◽

Vol 32 (suppl_1) ◽

Author(s):

Wakako Takabe ◽

Chih-Wen Ni ◽

Dong Ju Son ◽

Noah Alberts-Grill ◽

Hanjoong Jo

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Critical Role ◽

Oscillatory Shear ◽

Lim Domain ◽

Disturbed Flow ◽

Endothelial Inflammation ◽

And Migration ◽

Oscillatory Shear Stress ◽

Stable Flow

Recently, we have shown that disturbed flow, characterized by low and oscillatory shear stress, caused by a partial ligation of mouse left carotid artery (LCA) rapidly induces atherosclerosis. Using the partial ligation model and genome-wide microarray study with aortic endothelial RNAs obtained directly from the flow-disturbed carotid arteries, we previously identified mechanosensitive genes in mouse endothelial RNA including LIM domain only 4 ( lmo4 ). Here we report that LMO4 is a shear-sensitive protein that regulates endothelial inflammation. Lmo4 was up-regulated by disturbed flow in mouse LCA compared to the contralateral right CA (RCA) exposed to stable flow. At protein levels, LMO4 expression was significantly higher not only in LCA in our surgical model but also in the lesser curvature (flow-disturbed and athero-prone region of mouse aortic arch) compared to the greater curvature (stable-flow and ather-protected region). In addition, immunohistochemical staining of LMO4 in human coronary arteries revealed that its expression is detectable only in intimal endothelial cells, but not in medial cells. While LMO4 is known as a potential oncogene and associated with growth, migration and invasion of breast cancer cells, its role in cardiovascular system is not known to our knowledge. We tested a hypothesis that LMO4 is a mechanosensitive gene and plays a critical role in regulation of endothelial cell biology. LMO4 protein expression was robustly induced by oscillatory shear stress (OS) compared to laminar shear (LS) in human umbilical vein endothelial cells (HUVEC). Treatment of HUVEC with siRNA against LMO4 significantly inhibited OS-induced inflammation and migration, but not apoptosis and cell cycle progression. Further, LMO4 siRNA treatment significantly blunted expression of VCAM-1 and interleukin-8 induced by OS in endothelial cells. These results suggest that LMO4 is a shear-induced gene that plays a critical role in OS-induced endothelial inflammation and migration, and potentially in atherosclerosis.

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3-D Simulation for Blood Flow and Artery Compression in Asymmetric Stenotic Arteries With Axial Stretch

10.1115/imece2001/bed-23124 ◽

2001 ◽

Author(s):

Dalin Tang ◽

Chun Yang ◽

Shunnichi Kobayashi

Keyword(s):

Experimental Data ◽

Mechanical Properties ◽

Shear Stress ◽

Wall Stress ◽

Small Strain ◽

Rigid Wall ◽

Severe Stenosis ◽

Physiological Conditions ◽

Oscillating Shear Stress ◽

Stenotic Arteries

Abstract There has been increasing evidence that severe stenosis may cause artery compression and plaque cap rupture leading to heart attack and stroke. The physiological conditions under which that may occur and mechanisms involved are not well understood. It has been known that severe stenosis causes critical flow and wall mechanical conditions such as flow limitation, flow separation, low and oscillating shear stress distal to the stenosis, high shear stress and low or even negative flow pressure at the throat of stenosis, artery compression or even collapse. Those conditions are related to limitation of blood supply, intimal thickening and thrombosis formation, endothelism damage, platelet activation and aggregation, plaque cap rupture (for review, see [1,2]). Due to the complexity of the problem and lack of experimental data for mechanical properties of arteries under both expansion and compression, previous models were limited primarily to flow behaviors and with various limitations (axisymmetry, rigid wall, small strain, small pressure gradient). In this paper, experimental data for artery mechanical properties under physiological conditions were measured and a 3-d computational model is introduced to investigate flow behaviors and wall stress and strain distributions with fluid-structure interactions to better understand the mechanism involved in artery compression and plaque cap rupture.

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Effects of Caveolin-1-ERK1/2 pathway on endothelial cells and smooth muscle cells under shear stress

Experimental Biology and Medicine ◽

10.1177/1535370219892574 ◽

2019 ◽

Vol 245 (1) ◽

pp. 21-33 ◽

Cited By ~ 1

Author(s):

Lan Jia ◽

Lihua Wang ◽

Fang Wei ◽

Chen Li ◽

Zhe Wang ◽

...

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Smooth Muscle ◽

Growth Factor ◽

Caveolin 1 ◽

Low Shear Stress ◽

And Migration ◽

Oscillating Shear Stress ◽

Low Shear ◽

Proliferation And Migration

Hemodynamic forces have an important role in venous intimal hyperplasia, which is the main cause of arteriovenous fistula dysfunction. Endothelial cells (ECs) constantly exposed to the shear stress of blood flow, converted the mechanical stimuli into intracellular signals, and interacted with the underlying vascular smooth muscle cells (VSMCs). Caveolin-1 is one of the important mechanoreceptors on cytomembrane, which is related to vascular abnormalities. Extracellular signal-regulated kinase1/2 (ERK1/2) pathway is involved in the process of VSMCs proliferation and migration. In the present study, we explore the effects of Caveolin-1-ERK1/2 pathway and uremia toxins on the endothelial cells and VSMCs following shear stress application. Different shear stress was simulated with a ECs/VSMCs cocultured parallel-plate flow chamber system. Low shear stress and oscillating shear stress up-regulated the expression of fibroblast growth factor-4, platelet-derived growth factor-BB, vascular endothelial growth factor-A, ERK1/2 phosphorylation in endothelial cells, and proliferation and migration of VSMCs but down-regulated the Caveolin-1 expression in endothelial cells. Uremia toxin induces the proliferation and migration of VSMCs but not in a Caveolin-1-dependent manner in the static environment. Low shear stress-induced proliferation and migration of VSMCs is inhibited by Caveolin-1 overexpression and ERK1/2 suppression. Shear stress-regulated VSMC proliferation and migration is an endothelial cells-dependent process. Low shear stress and oscillating shear stress exert atherosclerotic influences on endothelial cells and VSMCs. Low shear stress modulated proliferation and migration of VSMCs through Caveolin-1-ERK1/2 pathway, which suggested that Caveolin-1 and ERK1/2 can be used as a new therapeutic target for the treatment of arteriovenous fistula dysfunction. Impact statement Venous intimal hyperplasia is the leading cause of arteriovenous fistula (AVF) dysfunction. This article reports that shear stress-regulated vascular smooth muscle cells (VSMCs) proliferation and migration is an endothelial cell (EC)-dependent process. Low shear stress (LSS) and oscillating shear stress (OSS) exert atherosclerotic influences on the ECs and VSMCs. LSS-induced proliferation and migration of VSMCs is inhibited by Caveolin-1 overexpression and extracellular signal-regulated kinase1/2 (ERK1/2) suppression, which suggested that Caveolin-1 and ERK1/2 can be used as a new therapeutic target for the treatment of AVF dysfunction.

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Exploring High Frequency Temporal Fluctuations in the Terminal Aneurysm of the Basilar Bifurcation

Journal of Biomechanical Engineering ◽

10.1115/1.4007279 ◽

2012 ◽

Vol 134 (9) ◽

Cited By ~ 15

Author(s):

Matthew D. Ford ◽

Ugo Piomelli

Keyword(s):

Shear Stress ◽

Wall Shear Stress ◽

High Frequency ◽

Cerebral Aneurysms ◽

Patient Specific ◽

Disturbed Flow ◽

Frequency Fluctuations ◽

Wall Shear ◽

Inflow Jet

Cerebral aneurysms are a common cause of death and disability. Of all the cardiovascular diseases, aneurysms are perhaps the most strongly linked with the local fluid mechanic environment. Aside from early in vivo clinical work that hinted at the possibility of high-frequency intra-aneurysmal velocity oscillations, flow in cerebral aneurysms is most often assumed to be laminar. This work investigates, through the use of numerical simulations, the potential for disturbed flow to exist in the terminal aneurysm of the basilar bifurcation. The nature of the disturbed flow is explored using a series of four idealized basilar tip models, and the results supported by four patient specific terminal basilar tip aneurysms. All four idealized models demonstrated instability in the inflow jet through high frequency fluctuations in the velocity and the pressure at approximately 120 Hz. The instability arises through a breakdown of the inflow jet, which begins to oscillate upon entering the aneurysm. The wall shear stress undergoes similar high-frequency oscillations in both magnitude and direction. The neck and dome regions of the aneurysm present 180 deg changes in the direction of the wall shear stress, due to the formation of small recirculation zones near the shear layer of the jet (at the frequency of the inflow jet oscillation) and the oscillation of the impingement zone on the dome of the aneurysm, respectively. Similar results were observed in the patient-specific models, which showed high frequency fluctuations at approximately 112 Hz in two of the four models and oscillations in the magnitude and direction of the wall shear stress. These results demonstrate that there is potential for disturbed laminar unsteady flow in the terminal aneurysm of the basilar bifurcation. The instabilities appear similar to the first instability mode of a free round jet.

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Oscillating shear stress mediates mesenchymal transdifferentiation of EPCs by the Kir2.1 channel

Heart and Vessels ◽

10.1007/s00380-020-01625-w ◽

2020 ◽

Vol 35 (10) ◽

pp. 1473-1482 ◽

Cited By ~ 1

Author(s):

Jifeng Li ◽

Yanting He ◽

Hongnan Bu ◽

Meiyue Wang ◽

Jie Yu ◽

...

Keyword(s):

Shear Stress ◽

Oscillating Shear Stress ◽

Kir2.1 Channel

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The Adaptor Protein Nck1, but not Nck2, Mediates Shear Stress-Induced Endothelial Permeability

10.1101/651687 ◽

2019 ◽

Author(s):

Mabruka Alfaidi ◽

Umesh Bhattarai ◽

Elizabeth D Cockerham ◽

A.W. Orr

Keyword(s):

Shear Stress ◽

Vascular Permeability ◽

Pore Formation ◽

Critical Role ◽

Endothelial Permeability ◽

Adaptor Protein ◽

Adaptor Proteins ◽

Disturbed Flow

AbstractAlteration in hemodynamic shear stress at atheroprone sites promotes endothelial paracellular pore formation and permeability. Previously, we have reported that a peptide inhibitor to Nck prevented shear stress-induced p21 activated kinase (PAK) activation and endothelial permeability. However, the specificity of this peptide is unclear, and the role of individual Nck isoforms remain unknown. Here, we show that genetic deletion of Nck1/2 adaptor proteins significantly ameliorates shear stress induced permeability, and selective isoform depletion suggests distinct signaling mechanisms. Only Nck1 deletion significantly reduces flow-induced paracellular pore formation and permeability, whereas Nck2 depletion has no significant effects. Additionally, Nck1 reexpression, but not Nck2, restores shear stress-induced permeability in Nck1/2 knockout cells, confirming the non-compensating roles. In vivo, using the partial carotid ligation model of disturbed flow, Nck1 knockout prevented the increase in vascular permeability, as assessed by both Evans blue extravasation and leakage of plasma fibrinogen into the vessel wall. Domain swap experiments mixing SH2 (phosphotyrosine binding) and SH3 (proline rich binding) domains between Nck1 and Nck2 showed a dispensable role for SH2 domains but a critical role for the Nck1 SH3 domains in rescuing shear stress-induced endothelial permeability. Consistent with this, both Nck1 and Nck2 bind to PECAM-1 (SH2 dependent) in response to shear stress, but only Nck1 ablation interferes with shear stress-induced PAK2 activation (SH3 dependent). This work provides the first evidence that Nck1 and Nck2 play distinct roles in flow-induced vascular permeability.New and NoteworthyThe present study shows a specific role for Nck1 in endothelial permeability in response to shear stress. Using in vitro and in vivo models, we demonstrate improvement in endothelial barrier integrity in cells subjected to disturbed flow only following Nck1 but not Nck2 deletion. Selective Nck1 inhibition may limit endothelial permeability at sites of disturbed flow to reduce atherosclerosis without affecting angiogenesis, which requires both Nck1 and Nck2 inhibition.

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Shear Stress Alterations Activate BMP4/pSMAD5 Signaling and Induce Endothelial Mesenchymal Transition in Varicose Veins

Cells ◽

10.3390/cells10123563 ◽

2021 ◽

Vol 10 (12) ◽

pp. 3563

Author(s):

Karthika Chandran Latha ◽

Ahalya Sreekumar ◽

Vyshna Beena ◽

Binil Raj S.S. ◽

RaviKumar B. Lakkappa ◽

...

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Venous Insufficiency ◽

Varicose Veins ◽

Disturbed Flow ◽

Mesenchymal Transition ◽

Venous Diseases ◽

Low Shear Stress ◽

Low Shear

Chronic venous diseases, including varicose veins, are characterized by hemodynamic disturbances due to valve defects, venous insufficiency, and orthostatism. Veins are physiologically low shear stress systems, and how altered hemodynamics drives focal endothelial dysfunction and causes venous remodeling is unknown. Here we demonstrate the occurrence of endothelial to mesenchymal transition (EndMT) in human varicose veins. Moreover, the BMP4-pSMAD5 pathway was robustly upregulated in varicose veins. In vitro flow-based assays using human vein, endothelial cells cultured in microfluidic chambers show that even minimal disturbances in shear stress as may occur in early stages of venous insufficiency induce BMP4-pSMAD5-based phenotype switching. Furthermore, low shear stress at uniform laminar pattern does not induce EndMT in venous endothelial cells. Targeting the BMP4-pSMAD5 pathway with small molecule inhibitor LDN193189 reduced SNAI1/2 expression in venous endothelial cells exposed to disturbed flow. TGFβ inhibitor SB505124 was less efficient in inhibiting EndMT in venous endothelial cells exposed to disturbed flow. We conclude that disturbed shear stress, even in the absence of any oscillatory flow, induces EndMT in varicose veins via activation of BMP4/pSMAD5-SNAI1/2 signaling. The present findings serve as a rationale for the possible use of small molecular mechanotherapeutics in the management of varicose veins.

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Abstract 515: Comparison of Aging and Disturbed Flow Effects on Arterial Wall Biomechanics

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.36.suppl_1.515 ◽

2016 ◽

Vol 36 (suppl_1) ◽

Author(s):

Anastassia Pokutta-Paskaleva ◽

Rudolph L Gleason ◽

Luke P Brewster

Keyword(s):

Shear Stress ◽

Carotid Artery ◽

Arterial Stiffness ◽

Arterial Compliance ◽

Confocal Imaging ◽

Disturbed Flow ◽

Arterial Stiffening ◽

Post Surgery ◽

Age Related ◽

Old Mice

Introduction: Arterial stiffness is an age related disease that doubles ones cardiovascular mortality and functions as an intermediary step in the formation of atherosclerosis. We have recently demonstrated that low and oscillatory wall shear stress, termed disturbed flow (d-flow), leads to arterial stiffness in otherwise normal arteries. Since d-flow has been linked to atherosclerotic plaque formation, our objective is to compare the carotid artery mechanics of 12 week arteries exposed to 4 weeks of d-flow to that of untreated 80 week old s129 mice. We hypothesize that low and oscillatory shear governs the remodeling of the arteries in a way that mimics the effect of arterial stiffening associated with aging. Materials and Methods: We used a partial carotid ligation model in 12-week-old s129 male (n=3) and female (n=3) mice. 4 weeks post surgery we euthanized the animals and performed biaxial extension-inflation biomechanical testing, confocal imaging, histological studies and opening angle studies on the left (partially ligated) carotid artery (LCA) and right (non-ligated) carotid artery (RCA) in order to characterize their biomechanical behavior. The same testing was performed on s129 80-week-old male animals (n=4). Pressure-diameter (P-d) data were collected from cyclic pressurization ramps from 0 to 160 mmHg at constant axial extension levels ranging from 1.3 to 1.9. Compliance was calculated at the in-vivo axial stretch level of 1.7 for both age groups. Intima media thickness was analyzed from H&E stained histological slides. Results were compared with the unpaired, two-tailed t-test and significance was taken at p<0.05. Results and Conclusions: Partial ligation for 4 weeks drastically decreased the compliance of the 12-week-old left carotid in a way that exactly follows the stiffening of the arteries in the 80-week-old mice. The compliance of the non-manipulated right carotid arteries reflected the age related difference in the stiffness of young vs old arteries with statistical difference in the 30 - 90 mmHg range. We have established that low shear stress and oscillatory flow promote the increase in arterial stiffening in the partially ligated young mice in a way that mimics the age-related decrease in arterial compliance in old mice.

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