Effect of Continuous Suppression of Heparan Sulfate during Flow on Arterial Endothelial Function
Ontology highlight
ABSTRACT:
ORGANISM(S): Sus scrofa
PROVIDER: GSE30843 | GEO | 2011/07/22
SECONDARY ACCESSION(S): PRJNA144155
REPOSITORIES: GEO
Similar Datasets
Project description:The arterial endothelium’s response to its flow environment is critical to vascular homeostasis. The endothelial glycocalyx has been shown to play a major role in mechanotransduction, but the extent to which the components of the glycocalyx affect the overall function of the endothelium remains unclear. The objective of this study was to further elucidate the role of heparan sulfate as a mechanosensor on the surface of the arterial endothelium, by (1) expanding the variety of shear waveforms investigated, (2) continuously suppressing heparan sulfate expression rather than using a pre-flow batch treatment, and (3) performing microarray analysis on post-flow samples. Porcine aortic endothelial cells were exposed to non-reversing, reversing, and oscillatory shear waveforms for 24 hours with or without continuous heparan sulfate suppression with heparinase. All shear waveforms significantly increased the amount of heparan sulfate on the surface of the endothelium. Suppression of heparan sulfate to less than 25% of control levels did not inhibit shear-induced cell alignment or nitric oxide production, or alter gene expression, for any of the shear waveforms investigated. We infer that heparan sulfate on the surface of porcine aortic endothelial cells is not the primary mechanosensor for many shear-responsive endothelial cell functions in this species. Porcine aortic endothelial cells were exposed to 3 different shear waveforms for 24 hours with or without the addition of 300 mU/ml heparinase III to the flow media. The shear waveforms inculded Non-reversing (15 ± 15 dyne/cm2, 1 Hz), Steady (15 dyne/cm2), or Oscillatory (0 ± 15 dyne/cm2, 1 Hz) shear. Four replicates of each condition were performed for a total of 24 experiments. Each experimental sample was hybridized to an oligonucleotide array along with a standard reference sample (static cells).
2011-07-21 | E-GEOD-30843 | biostudies-arrayexpress
Project description:Giantsos-Adams2013 - Growth of glycocalyx
under static conditions
This model is described in the article:
Heparan Sulfate Regrowth
Profiles Under Laminar Shear Flow Following Enzymatic
Degradation.
Giantsos-Adams KM, Koo AJ, Song S,
Sakai J, Sankaran J, Shin JH, Garcia-Cardena G, Dewey CF.
Cell Mol Bioeng 2013 Jun; 6(2):
160-174
Abstract:
The local hemodynamic shear stress waveforms present in an
artery dictate the endothelial cell phenotype. The observed
decrease of the apical glycocalyx layer on the endothelium in
atheroprone regions of the circulation suggests that the
glycocalyx may have a central role in determining
atherosclerotic plaque formation. However, the kinetics for the
cells' ability to adapt its glycocalyx to the environment have
not been quantitatively resolved. Here we report that the
heparan sulfate component of the glycocalyx of HUVECs increases
by 1.4-fold following the onset of high shear stress, compared
to static cultured cells, with a time constant of 19 h.
Cell morphology experiments show that 12 h are required
for the cells to elongate, but only after 36 h have the
cells reached maximal alignment to the flow vector. Our
findings demonstrate that following enzymatic degradation,
heparan sulfate is restored to the cell surface within
12 h under flow whereas the time required is 20 h
under static conditions. We also propose a model describing the
contribution of endocytosis and exocytosis to apical heparan
sulfate expression. The change in HS regrowth kinetics from
static to high-shear EC phenotype implies a differential in the
rate of endocytic and exocytic membrane turnover.
This model is hosted on
BioModels Database
and identified by:
MODEL1609100001.
To cite BioModels Database, please use:
BioModels Database:
An enhanced, curated and annotated resource for published
quantitative kinetic models.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
2016-09-28 | MODEL1609100001 | BioModels
Project description:Giantsos-Adams2013 - Growth of glycocalyx
under static conditions
This model is described in the article:
Heparan Sulfate Regrowth
Profiles Under Laminar Shear Flow Following Enzymatic
Degradation.
Giantsos-Adams KM, Koo AJ, Song S,
Sakai J, Sankaran J, Shin JH, Garcia-Cardena G, Dewey CF.
Cell Mol Bioeng 2013 Jun; 6(2):
160-174
Abstract:
The local hemodynamic shear stress waveforms present in an
artery dictate the endothelial cell phenotype. The observed
decrease of the apical glycocalyx layer on the endothelium in
atheroprone regions of the circulation suggests that the
glycocalyx may have a central role in determining
atherosclerotic plaque formation. However, the kinetics for the
cells' ability to adapt its glycocalyx to the environment have
not been quantitatively resolved. Here we report that the
heparan sulfate component of the glycocalyx of HUVECs increases
by 1.4-fold following the onset of high shear stress, compared
to static cultured cells, with a time constant of 19 h.
Cell morphology experiments show that 12 h are required
for the cells to elongate, but only after 36 h have the
cells reached maximal alignment to the flow vector. Our
findings demonstrate that following enzymatic degradation,
heparan sulfate is restored to the cell surface within
12 h under flow whereas the time required is 20 h
under static conditions. We also propose a model describing the
contribution of endocytosis and exocytosis to apical heparan
sulfate expression. The change in HS regrowth kinetics from
static to high-shear EC phenotype implies a differential in the
rate of endocytic and exocytic membrane turnover.
This model is hosted on
BioModels Database
and identified by:
MODEL1609100001.
To cite BioModels Database, please use:
BioModels Database:
An enhanced, curated and annotated resource for published
quantitative kinetic models.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
2019-10-09 | BIOMD0000000830 | BioModels
Project description:Sepsis patients are at increased risk for hospital-acquired pulmonary infections, potentially due to post-septic immunosuppression known as the compensatory anti-inflammatory response syndrome (CARS). CARS has been attributed to leukocyte dysfunction, with an unclear role for endothelial cells. The pulmonary circulation is lined by an endothelial glycocalyx, a heparan sulfate-rich layer essential to pulmonary homeostasis. Heparan sulfate degradation occurs early in sepsis, leading to lung injury. Endothelial synthesis of new heparan sulfates subsequently allows for glycocalyx reconstitution and endothelial recovery. We hypothesized that remodeling of the reconstituted endothelial glycocalyx, mediated by alterations in the endothelial machinery responsible for heparan sulfate synthesis, contributes to CARS. Our experimental animal model of CARS recapitulated post-septic immunosuppression, coincidentally with structural remodeling of endothelial glycocalyx heparan sulfate. We used microarray to identify which heparan sulfate modifying enzyme is responsible for the remodeling of post-septic reconstituted glycocalyx, characterized with enrichment of heparan sulfate disaccharides sulfated at the 6-O position of glucosamine.
2019-04-14 | GSE129775 | GEO
Project description:Human induced pluripotent stem cells (hiPSCs) are used to study organogenesis and model disease as well as being developed for regenerative medicine. Endothelial cells are among the many cell types differentiated from hiPSC, but their maturation and stabilization fall short of that in adult endothelium. We examined whether shear stress alone or in combination with pericyte co-culture would induce flow alignment and maturation of hiPSC-derived endothelial cells (hiPSC-ECs) but found no effects comparable to those in primary microvascular EC. In addition, hiPSC-ECs lacked a luminal glycocalyx, critical for vasculature homeostasis, shear stress sensing and signalling. We noted however, that hiPSC-EC have dysfunctional mitochondrial permeability transition pores, resulting in reduced mitochondrial function and increased reactive oxygen species (ROS). Closure of these pores by cyclosporine-A improved EC mitochondrial function but also restored the glycocalyx such that alignment to flow took place. These indicated that mitochondrial maturation is required for proper hiPSC-EC functionality.
2019-12-14 | E-MTAB-8392 | biostudies-arrayexpress
Project description:Vasculature permeate our entire body and involved in homeostasis and progression of various disease. Endothelium is a backbone of cardiovascular system and endothelial disfunction is associated with most forms of cardiovascular disease from atherosclerosis to chronic heart failure. Decrease in shear stress, particularly due to disturbance of the blood flow by bends or vascular obstruction, lead to atherosclerosis and endothelial disfunction. Notch signaling may act as mechanosensor and assumed to be one of the central signal pathways associated with endothelial disfunction. Association of shear stress, Notch and endothelial dysfunction is well known, molecular mechanisms connected these factors still poorly understood. One might assume that long-term shear stress and dysregulation of Notch in endothelium cause changes in epigenomic profile. While epigenomic changes in shear stressed environment are known to be involved in endothelial disfunction, possible connection of Notch and epigenetic changes in endothelium has not yet been tested. Therefore, here we analyzed how activation of Notch signaling influence on histone code and secretome of endothelial cells in vitro. We found that activation of Notch increase level of N-acetylated forms of Histone 1: H1-0, H1-3, H1-4, H1-5, H1-10. These changes were also associated with changes in secretome profile of endothelium and might have broad biomedical significance in case of endothelial dysfunction.
2023-02-15 | PXD032978 | Pride
Project description:The highly negatively charged endothelial surface glycocalyx (ESG) functions as mechano-sensor detecting shear forces generated by the blood flow on the luminal side of brain endothelial cells (ECs) and contributes to the physical barrier of the blood-brain barrier (BBB). Despite the importance of ESG in the regulation of BBB permeability in physiological conditions and in diseases, this is an underresearched area. Microfluidic lab-on-a-chip (LOC) devices allow the study of BBB properties in dynamic conditions. We studied a BBB model, human endothelial cells derived from hematopoetic stem cells in co-culture with brain pericytes, in an LOC device to understand the role of fluid flow in the regulation of ESG-related genes and surface charge. The MACE gene sequencing study showed differentially expressed core protein genes of the ESG after fluid flow, as well as enriched pathways for the extracellular matrix molecules. We observed increased barrier properties, a higher intensity glycocalyx staining and a more negative surface charge of human brain ECs in dynamic conditions. Our study is the first to provide data on ESG of human ECs in an LOC device under dynamic conditions and confirm the importance of fluid flow for BBB culture models.
2021-09-01 | GSE155671 | GEO
Project description:The lymphatic system removes fluid from the interstitial space and returns it to the blood with a tremendous capacity: during inflammation, lymph flow rates can increase dramatically; however, during chronic lymphedema, there is little or no flow. The ability of lymphatic endothelium to sense and actively regulate this function is unknown, and shear stress is likely a key indicator of lymph flow. We profiled gene expression in human dermal microvascular lymphatic endothelial cells exposed to 0, 2 and 20 dyn/cm2 shear stress as representative of chronic lymphedema, normal, and acute inflammatory conditions, respectively. We found important adaptive responses correlated to multiple aspects of lymphatic function. Importantly, shear stress upregulated intracellular water and solute transporters while decreasing cell-cell adhesion and basement membrane components and increasing cell-matrix interactions. This data indicate that during high loading conditions, both passive and active drainage function increases, while conversely when fluid drainage is blocked, transport function is diminished in the lymphatic endothelium. These data demonstrate the first functional-adaptive response of lymphatic endothelium to flow conditions, thus indicating that the lymphatic endothelium plays an active role in regulating their function. Keywords: Shear stress, dose response, cell type comparison Lymphatic endothelial cells were subjected to 0, 2, or 20 dyn/cm2 shear stress; blood endothelial cells were subjected to 0 or 20 dyn/cm2 shear stress. Four samples were used for each cell type/shear level group for a total of 20 samples. Each sample was independently compared to human universal reference RNA via two-color microarray analysis for a total of 20 arrays. In all cases, the experimental samples were labeled with Cy5 dye while the reference RNA was labeled with Cy3.
2009-11-30 | E-GEOD-8852 | biostudies-arrayexpress
Project description:Infiltration of peripheral immune cells after blood-brain barrier (BBB) dysfunction causes severe inflammation after a stroke. Although the endothelial glycocalyx functions as a barrier to circulating cells, the relationship between stroke severity and glycocalyx dysfunction remains unclear. In this study, glycosaminoglycans (GAGs), a component of the endothelial glycocalyx, in ischemic stroke were studied using a photochemically induced thrombosis (PIT) mouse model. As results, decreased levels of heparan sulfate (HS) and chondroitin sulfate (CS) and increased activity of hyaluronidase 1 and heparanase (HPSE) were observed in ischemic brain tissues. HPSE expression in cerebral vessels increased after stroke onset and infarct volume greatly decreased after co-administration of N-acetylcysteine (NAC)+GAG oligosaccharides as compared to NAC administration alone. These results suggest that the endothelial glycocalyx was injured after the onset of stroke. Interestingly, scission activity of proHPSE produced by HBMEC/ciβ and HEK293 cells transfected with hHPSE1 cDNA were activated by acrolein exposure. We identified the ACR modified amino acid residues of proHPSE using nano LC-MS/MS, suggesting that ACR modification of Lys139 (6-kDa linker), and Lys107 and Lys161, located in the immediate vicinity of the 6-kDa linker, at least in part, is attributed to the activation of proHPSE. Since proHPSE, but not HPSE, localizes outside cells by binding with HS proteoglycans, ACR-modified proHPSE represents a promising target to protect the endothelial glycocalyx.
2020-11-16 | PXD021040 | Pride
Project description:Hutchinson-Gilford Progeria Syndrome (HGPS) is a premature aging disease in children that leads to early death. Smooth muscle cells (SMCs) are the most affected cells in HGPS patients, although the reason for such vulnerability remains poorly understood. In this work, we developed a chip formed from HGPS-SMCs that were generated from induced pluripotent stem cells (iPSCs) to study their vulnerability to flow shear stress. HGPS-iPSC SMCs cultured under arterial flow conditions detached from the chip after a few days of culture; this process was mediated by the up-regulation of metalloprotease 13 (MMP13). Importantly, double mutant LmnaG609G/G609GMmp13-/- mice or LmnaG609G/G609GMmp13+/+ mice treated with a MMP inhibitor showed lower SMC loss in the aortic arch than controls. MMP13 up-regulation appears to be mediated by the up-regulation of heparan sulfate, a glycocalyx component. Our results offer a new platform for developing treatments for HGPS patients that may complement previous pre-clinical and clinical treatments.
2020-08-24 | PXD011652 | Pride