scholarly journals DNA motif analysis of shear stress responsive endothelial enhancers reveals differential association of KLF and ETV/ETS binding sites with gained and lost enhancers

2021 ◽  
Author(s):  
Arndt F Siekmann ◽  
Roman Tsaryk ◽  
Nora Yucel ◽  
Zoltan Arany ◽  
Olga Bondareva ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Binding Sites ◽  
Regulatory Elements ◽  
Open Chromatin ◽  
Laminar Shear Stress ◽  
Rna Seq ◽  
Motif Analysis ◽  
Dna Motif ◽  
Laminar Shear

Endothelial cells (EC) lining blood vessels are exposed to mechanical forces, such as shear stress exerted by the flowing blood. These forces control many aspects of EC biology, including vascular tone, cell migration and proliferation in addition to cell size and shape. Despite a good understanding of the genes and signaling pathways responding to shear stress, our insights into the transcriptional regulation of these responses is much more limited. In particular, we do not know the different sets of regulatory elements (enhancers) that might control increases or decreases in gene expression. Here, we set out to study changes in the chromatin landscape of human umbilical vein endothelial cells (HUVEC) exposed to laminar shear stress. To do so, we performed ChIP-Seq for H3K27 acetylation, indicative of active enhancer elements and ATAC-Seq to mark regions of open chromatin in addition to RNA-Seq on HUVEC exposed to 6 hours of laminar shear stress. Our results show a correlation of gained and lost enhancers with up- and downregulated genes, respectively. DNA motif analysis revealed an over-representation of KLF transcription factor (TF) binding sites in gained enhancers, while lost enhancers contained more ETV/ETS motifs. We validated a subset of flow responsive enhancers using luciferase-based reporter constructs and CRISPR-Cas9 mediated genome editing. Lastly, we characterized shear stress responsive genes in ECs of zebrafish embryos using RNA-Seq. Together, our results reveal the presence of shear stress responsive DNA regulatory elements and lay the groundwork for the future exploration of these elements and the TFs binding to them in controlling EC biology.

scholarly journals Primary cilia of human endothelial cells disassemble under laminar shear stress

2004 ◽  
Vol 164 (6) ◽  
pp. 811-817 ◽  
Author(s):  
Carlo Iomini ◽  
Karla Tejada ◽  
Wenjun Mo ◽  
Heikki Vaananen ◽  
Gianni Piperno
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Primary Cilia ◽  
Umbilical Vein ◽  
Intraflagellar Transport ◽  
Mechanical Stimuli ◽  
Laminar Shear Stress ◽  
Human Endothelial Cells ◽  
Umbilical Vein Endothelial Cells ◽  
Laminar Shear

We identified primary cilia and centrosomes in cultured human umbilical vein endothelial cells (HUVEC) by antibodies to acetyl-α-tubulin and capillary morphogenesis gene-1 product (CMG-1), a human homologue of the intraflagellar transport (IFT) protein IFT-71 in Chlamydomonas. CMG-1 was present in particles along primary cilia of HUVEC at interphase and around the oldest basal body/centriole at interphase and mitosis. To study the response of primary cilia and centrosomes to mechanical stimuli, we exposed cultured HUVEC to laminar shear stress (LSS). Under LSS, all primary cilia disassembled, and centrosomes were deprived of CMG-1. We conclude that the exposure to LSS ends the IFT in cultured endothelial cells.

Identification of atheroprone shear stress responsive regulatory elements in endothelial cells

2019 ◽  
Vol 115 (10) ◽  
pp. 1487-1499 ◽  
Author(s):  
Olga Bondareva ◽  
Roman Tsaryk ◽  
Vesna Bojovic ◽  
Maria Odenthal-Schnittler ◽  
Arndt F Siekmann ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Umbilical Vein ◽  
Regulatory Element ◽  
Hippo Pathway ◽  
Regulatory Elements ◽  
Binding Motif ◽  
Motif Analysis ◽  
Static Conditions

Abstract Aims Oscillatory shear stress (OSS) is an atheroprone haemodynamic force that occurs in areas of vessel irregularities and is implicated in the pathogenesis of atherosclerosis. Changes in signalling and transcriptional programme in response to OSS have been vigorously studied; however, the underlying changes in the chromatin landscape controlling transcription remain to be elucidated. Here, we investigated the changes in the regulatory element (RE) landscape of endothelial cells under atheroprone OSS conditions in an in vitro model. Methods and results Analyses of H3K27ac chromatin immunoprecipitation-Seq enrichment and RNA-Seq in primary human umbilical vein endothelial cells 6 h after onset of OSS identified 2806 differential responsive REs and 33 differentially expressed genes compared with control cells kept under static conditions. Furthermore, gene ontology analyses of putative RE-associated genes uncovered enrichment of WNT/HIPPO pathway and cytoskeleton reorganization signatures. Transcription factor (TF) binding motif analysis within RE sequences identified over-representation of ETS, Zinc finger, and activator protein 1 TF families that regulate cell cycle, proliferation, and apoptosis, implicating them in the development of atherosclerosis. Importantly, we confirmed the activation of EGR1 as well as the YAP/TAZ complex early (6 h) after onset of OSS in both cultured human vein and artery endothelial cells and, by undertaking luciferase assays, functionally verified their role in RE activation in response to OSS. Conclusions Based on the identification and verification of specific responsive REs early upon OSS exposure, we propose an expanded mechanism of how OSS might contribute to the development of atherosclerosis.

Laminar shear stress promotes mitochondrial homeostasis in endothelial cells

2018 ◽  
Vol 233 (6) ◽  
pp. 5058-5069 ◽  
Author(s):  
Li-Hong Wu ◽  
Hao-Chun Chang ◽  
Pei-Ching Ting ◽  
Danny L. Wang
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Laminar Shear Stress ◽  
Mitochondrial Homeostasis ◽  
Laminar Shear

A Promoter Polymorphism Regulates NFKB1 Gene Transactivity in Human Endothelial Cells under Laminar Shear Stress

2006 ◽  
Vol 38 (Supplement) ◽  
pp. S4
Author(s):  
Joon Y. Park ◽  
Iain K. Farrance ◽  
Hanjoong Jo ◽  
Steven R. Brant ◽  
Stephen M. Roth ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Promoter Polymorphism ◽  
Laminar Shear Stress ◽  
Human Endothelial Cells ◽  
Laminar Shear

AXL ACTIVATION MEDIATES LAMINAR SHEAR STRESS ANTIAPOPTOTIC EFFECTS IN HUMAN ENDOTHELIAL CELLS

2004 ◽  
Vol 13 (3) ◽  
pp. 194
Author(s):  
Daniela D'Arcangelo ◽  
Valeria Ambrosino ◽  
Gianluca Ragone ◽  
Maria Giannuzzo ◽  
Maurizio C Capogrossi ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Laminar Shear Stress ◽  
Human Endothelial Cells ◽  
Laminar Shear

Laminar shear stress inhibits CXCR4 expression on endothelial cells: functional consequences for atherogenesis

2005 ◽  
Vol 19 (6) ◽  
pp. 1-25 ◽  
Author(s):  
Roberta Melchionna ◽  
Daniele Porcelli ◽  
Antonella Mangoni ◽  
Daniele Carlini ◽  
Giovanna Liuzzo ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Cxcr4 Expression ◽  
Laminar Shear Stress ◽  
Functional Consequences ◽  
Laminar Shear

Recruitment of endothelial caveolae into mechanotransduction pathways by flow conditioning in vitro

2003 ◽  
Vol 285 (4) ◽  
pp. H1720-H1729 ◽  
Author(s):  
Victor Rizzo ◽  
Christine Morton ◽  
Natacha DePaola ◽  
Jan E. Schnitzer ◽  
Peter F. Davies
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Cell Surface ◽  
Tyrosine Phosphorylation ◽  
Surface Proteins ◽  
Luminal Surface ◽  
Laminar Shear Stress ◽  
Caveolin 1 ◽  
Cultured Endothelial Cells ◽  
Laminar Shear

The luminal surface of rat lung microvascular endothelial cells in situ is sensitive to changing hemodynamic parameters. Acute mechanosignaling events initiated in response to flow changes in perfused lung microvessels are localized within specialized invaginated microdomains called caveolae. Here we report that chronic exposure to shear stress alters caveolin expression and distribution, increases caveolae density, and leads to enhanced mechanosensitivity to subsequent changes in hemodynamic forces within cultured endothelial cells. Flow-preconditioned cells expressed a fivefold increase in caveolin (and other caveolar-residing proteins) at the luminal surface compared with no-flow controls. The density of morphologically identifiable caveolae was enhanced sixfold at the luminal cell surface of flow-conditioned cells. Laminar shear stress applied to static endothelial cultures (flow step of 5 dyn/cm2), enhanced the tyrosine phosphorylation of luminal surface proteins by 1.7-fold, including caveolin-1 by 1.3-fold, increased Ser1179 phosphorylation of endothelial nitric oxide synthase (eNOS) by 2.6-fold, and induced a 1.4-fold activation of mitogen-activated protein kinases (ERK1/2) over no-flow controls. The same shear step applied to endothelial cells preconditioned under 10 dyn/cm2 of laminar shear stress for 6 h and induced a sevenfold increase of total phosphotyrosine signal at the luminal endothelial cell surface enhanced caveolin-1 tyrosine phosphorylation 5.8-fold and eNOS phosphorylation by 3.3-fold over static control values. In addition, phosphorylated caveolin-1 and eNOS proteins were preferentially localized to caveolar microdomains. In contrast, ERK1/2 activation was not detected in conditioned cells after acute shear challenge. These data suggest that cultured endothelial cells respond to a sustained flow environment by directing caveolae to the cell surface where they serve to mediate, at least in part, mechanotransduction responses.

Sign in / Sign up

Export Citation Format

Share Document