Angiogenic Response of Endothelial Cells to Fibronectin

2012 ◽  
pp. 131-151 ◽  
Author(s):  
V. B. S. Kumar ◽  
R. I. Viji ◽  
M. S. Kiran ◽  
Perumana R. Sudhakaran
Keyword(s):  
Endothelial Cells ◽  
Angiogenic Response

A chromatin signature by the methyltransferase SETD7 regulates semaphorin-3G transcription and angiogenic response in diabetes: insights for personalized epigenetic therapies

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
S.A Mohammed ◽  
S Costantino ◽  
A Akhmedov ◽  
G Karsai ◽  
S Ambrosini ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Gastrocnemius Muscle ◽  
Tube Formation ◽  
Public Institution ◽  
Epigenetic Therapy ◽  
Great Promise ◽  
Funding Source ◽  
Rna Seq ◽  
Angiogenic Response ◽  
Pharmacological Inhibition

Abstract Background Despite advances in revascularization strategies, type 2 diabetic (T2D) patients with peripheral artery disease (PAD) continue to have a high risk of limb amputation. Modulation of blood vessel growth holds great promise for the treatment of PAD patients. Epigenetic modifications, namely histone post-translational modifications, have shown to regulate transcriptional programs implicated in the pathogenesis of cardiovascular disease. Aim To investigate the role of chromatin changes in regulating post-ischemic vascularization in experimental diabetes as well as in patients with T2D. Methods Experiments were performed in primary human aortic endothelial cells (HAECs), double-mutant leptin deficient mice (Lepdb/db) carrying a genetic deletion of the methyltransferase SETD7 (Setd7−/−Lepdb/db) as well as in gastrocnemius muscle samples from T2D patients with PAD and age-matched non-diabetic controls. Unbiased gene expression profiling was performed by RNA sequencing (RNA-seq) followed by Ingenuity Pathway Analysis (IPA). Pharmacological blockade of SETD7 was performed by using the selective inhibitor (R)-PFI-2. Scratch and tube formation assays were performed to investigate the impact of SETD7 on angiogenic response. Results RNA-seq in high glucose-treated HAECs revealed a profound upregulation of the methyltransferase SETD7 (fold change 2.8, p<0.001), an enzyme involved in mono-methylation of lysine 4 at histone 3 (H3K4me1). Both SETD7 gene silencing and pharmacological inhibition by (R)PFI-2 rescued hyperglycemia-induced impairment of HAECs migration and tube formation, while SETD7 overexpression blunted the angiogenic response. RNA-seq and Chromatin Immunoprecipitation (ChIP) assays showed that SETD7-dependent H3K4me1 regulates the transcription of the angiogenesis inhibitor semaphorin-3G (SEMA-3G). Increased SEMA-3G transcript was associated with enhanced secretion from HAECs. Co-immunofluorescence experiments showed that SEMA-3G blunts the angiogenic response by competing with VEGF receptors VEGFR/Neuropillin2. Moreover, SEMA-3G overexpression blunted migration and tube formation in SETD7-depleted HAECs. SETD7 and SEMA-3G were significantly upregulated in endothelial cells from Lepdb/db mice, whereas SEMA-3G transcription was blunted in Setd7−/−Lepdb/db animals. Consistently, endothelial sprouting was defective in aortas from Lepdb/db as compared to WT mice, whereas Setd7−/−Lepdb/db mice displayed a preserved angiogenic response. Of clinical relevance, SETD7/SEMA-3G axis was upregulated in gastrocnemius muscle specimens from T2D patients with PAD as compared with non-diabetic controls. Conclusion In HAECs, genetically modified mice and T2D patients we show that SETD7-dependent chromatin changes regulate SEMA-3G transcription and angiogenic response. Pharmacological inhibition of SETD7 may represent a novel epigenetic therapy to boost neovascularization in T2D patients with PAD. Funding Acknowledgement Type of funding source: Public Institution(s). Main funding source(s): University of Zurich/Universitätsspital Zürich

Urokinase receptor expression on human microvascular endothelial cells is increased by hypoxia: implications for capillary-like tube formation in a fibrin matrix

Blood ◽  
2000 ◽  
Vol 96 (8) ◽  
pp. 2775-2783 ◽  
Author(s):  
Marielle E. Kroon ◽  
Pieter Koolwijk ◽  
Bea van der Vecht ◽  
Victor W. M. van Hinsbergh
Keyword(s):  
Endothelial Cells ◽  
Tube Formation ◽  
Microvascular Endothelial Cells ◽  
Tubular Structures ◽  
Angiogenic Response ◽  
Fibrin Matrix ◽  
Hypoxic Conditions ◽  
Upa Receptor ◽  
Tnf Α ◽  
Blocking Antibodies

Abstract Hypoxia stimulates angiogenesis, the formation of new blood vessels. This study evaluates the direct effect of hypoxia (1% oxygen) on the angiogenic response of human microvascular endothelial cells (hMVECs) seeded on top of a 3-dimensional fibrin matrix. hMVECs stimulated with fibroblast growth factor–2 (FGF-2) or vascular endothelial growth factor (VEGF) together with tumor necrosis factor–α (TNF-α) formed 2- to 3-fold more tubular structures under hypoxic conditions than in normoxic (20% oxygen) conditions. In both conditions the in-growth of capillary-like tubular structures into fibrin required cell-bound urokinase-type plasminogen activator (uPA) and plasmin activities. The hypoxia-induced increase in tube formation was accompanied by a decrease in uPA accumulation in the conditioned medium. This decrease in uPA level was completely abolished by uPA receptor-blocking antibodies. During hypoxic culturing uPA receptor activity and messenger RNA (mRNA) were indeed increased. This increase and, as a consequence, an increase in plasmin formation contribute to the hypoxia-induced stimulation of tube formation. A possible contribution of VEGF-A to the increased formation under hypoxic conditions is unlikely because there was no increased VEGF-A expression detected under hypoxic conditions, and the hypoxia-induced tube formation by FGF-2 and TNF-α was not inhibited by soluble VEGFR-1 (sVEGFR-1), or by antibodies blocking VEGFR-2. Furthermore, although the αv-integrin subunit was enhanced by hypoxia, blocking antibodies against αvβ3- and αvβ5-integrins had no effect on hypoxia-induced tube formation. Hypoxia increases uPA association and the angiogenic response of human endothelial cells in a fibrin matrix; the increase in the uPA receptor is an important determinant in this process.

scholarly journals Induction of tumor inhibitory anti-angiogenic response through immunization with interferon Gamma primed placental endothelial cells: ValloVax™

2015 ◽  
Vol 13 (1) ◽  
pp. 90 ◽  
Author(s):  
Thomas E Ichim ◽  
Shuang Li ◽  
Hong Ma ◽  
Yuliya V Yurova ◽  
Julia S Szymanski ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Interferon Gamma ◽  
Angiogenic Response

Ultrasound-Induced Angiogenic Response in Endothelial Cells

2007 ◽  
Vol 33 (11) ◽  
pp. 1818-1829 ◽  
Author(s):  
Natalya Mizrahi ◽  
Dror Seliktar ◽  
Eitan Kimmel
Keyword(s):  
Endothelial Cells ◽  
Angiogenic Response

scholarly journals Hyperbaric oxygen induces a cytoprotective and angiogenic response in human microvascular endothelial cells

2009 ◽  
Vol 15 (4) ◽  
pp. 431-442 ◽  
Author(s):  
Cassandra A. Godman ◽  
Kousanee P. Chheda ◽  
Lawrence E. Hightower ◽  
George Perdrizet ◽  
Dong-Guk Shin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Hyperbaric Oxygen ◽  
Microvascular Endothelial Cells ◽  
Angiogenic Response ◽  
Microvascular Endothelial

scholarly journals ETS1, ELK1, and ETV4 Transcription Factors Regulate Angiopoietin-1 Signaling and the Angiogenic Response in Endothelial Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Sharon Harel ◽  
Veronica Sanchez ◽  
Alaa Moamer ◽  
Javier E. Sanchez-Galan ◽  
Mohammad N. Abid Hussein ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Transcription Factors ◽  
Endothelial Cell ◽  
Binding Sites ◽  
Umbilical Vein ◽  
Intracellular Localization ◽  
Mrna Levels ◽  
Mobility Shift ◽  
Angiogenic Response ◽  
Angiopoietin 1

BackgroundAngiopoietin-1 (Ang-1) is the main ligand of Tie-2 receptors. It promotes endothelial cell (EC) survival, migration, and differentiation. Little is known about the transcription factors (TFs) in ECs that are downstream from Tie-2 receptors.ObjectiveThe main objective of this study is to identify the roles of the ETS family of TFs in Ang-1 signaling and the angiogenic response.MethodsIn silico enrichment analyses that were designed to predict TF binding sites of the promotors of eighty-six Ang-1-upregulated genes showed significant enrichment of ETS1, ELK1, and ETV4 binding sites in ECs. Human umbilical vein endothelial cells (HUVECs) were exposed for different time periods to recombinant Ang-1 protein and mRNA levels of ETS1, ELK1, and ETV4 were measured with qPCR and intracellular localization of these transcription factors was assessed with immunofluorescence. Electrophoretic mobility shift assays and reporter assays were used to assess activation of ETS1, ELK1, and ETV4 in response to Ang-1 exposure. The functional roles of these TFs in Ang-1-induced endothelial cell survival, migration, differentiation, and gene regulation were evaluated by using a loss-of-function approach (transfection with siRNA oligos).ResultsAng-1 exposure increased ETS1 mRNA levels but had no effect on ELK1 or ETV4 levels. Immunostaining revealed that in control ECs, ETS1 has nuclear localization whereas ELK1 and ETV4 are localized to the nucleus and the cytosol. Ang-1 exposure increased nuclear intensity of ETS1 protein and enhanced nuclear mobilization of ELK1 and ETV4. Selective siRNA knockdown of ETS1, ELK1, and ETV4 showed that these TFs are required for Ang-1-induced EC survival and differentiation of cells, while ETS1 and ETV4 are required for Ang-1-induced EC migration. Moreover, ETS1, ELK1, and ETV4 knockdown inhibited Ang-1-induced upregulation of thirteen, eight, and nine pro-angiogenesis genes, respectively.ConclusionWe conclude that ETS1, ELK1, and ETV4 transcription factors play significant angiogenic roles in Ang-1 signaling in ECs.

The Integrin Co-Activator Kindlin-2 Plays a Critical Role In Angiogenesis and Blood Vessel Integrity

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4-4
Author(s):  
Elzbieta Pluskota ◽  
James J Dowling ◽  
Natalie Gordon ◽  
Jeffrey A Golden ◽  
Dorota Szpak ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Blood Vessels ◽  
Vascular Permeability ◽  
Conflicts Of Interest ◽  
Critical Role ◽  
Dorsal Skin ◽  
Integrin Activation ◽  
Angiogenic Response ◽  
Vascular Area

Abstract Abstract 4 Kindlin-2, a widely distributed cytoskeletal protein, has been implicated in integrin activation, and its absence is embryonically lethal in mice and causes severe developmental defects in zebrafish. Our previous study demonstrated that reduction of kindlin-2 levels in human endothelial cells results in defective adhesive and migratory responses, suggesting that kindlin-2 may be implicated in angiogenesis. We tested this hypothesis in the kindlin-2+/− mice utilizing murine RM1 prostate tumor and Matrigel implant models. Staining of tumor sections for EC with CD31 showed shorter and thinner blood vessels and reduced vascular area by 3.5-fold in tumors grown in kindlin-2+/− mice compared to WT mice (P=0.0186, n=6). The vessels that did form in the kindlin-2+/− mice were immature as they lacked smooth muscle cells and pericytes, had thinner basement membrane, and were leaky as evidenced by increased by 2-fold area for plasma-derived fibrin in tumor sections (P=0.0006, n=5). Consistent with the blunted angiogenic response and vascular leakiness in the kindlin-2+/− mice, the tumors grown in kindlin-2+/− animals had 2-fold larger necrotic core and were 2.5-fold smaller than those derived in WT mice (P=0.042, n=7). Also, the permeability of preexisting blood vessels in ear and dorsal skin of WT and kindlin-2+/− mice was compared after injection of Evans blue dye. Baseline permeability of vasculature in ear and dorsal skin was enhanced by ∼70–100 % in kindlin-2+/− mice as compared WT mice (P<0.01, n=6). Application of mustard oil, a proinflammatory stimulus, on ear skin or VEGF on dorsal skin, enhanced permeability in WT mice by ∼75–80 % (P<0.05, n=6) while it had a negligible effect in the kindlin-2+/− mice (P=0.159, n=6). The impaired angiogenic response in the kindlin-2+/− mice was confirmed in a Matrigel model where VEGF-induced angiogenesis was abnormal. CD31 staining showed that in contrast to the matrigel plugs derived from WT mice, those obtained from the kindlin-2+/− mice contained shorter and thinner vessels, and the few thicker CD31+ structures that did form in the kindlin-2+/− mice were disorganized. As a consequence of blood leakage from abnormal vessels the Matrigel implants collected from the kindlin-2+/− mice exhibited a 3.5-fold increase in hemoglobin content as compared to WT mice (P=0.0149, n=5).Bone marrow transplant experiments were performed and showed that bone marrow-derived cells did not significantly contribute to abnormal angiogenesis and enhanced vascular permeability observed in the kindlin-2+/− mice. Tumor vascular area, plasma-derived fibrin content, tumor growth and vascular permeability in ear and dorsal skin were similar in WT recipients, which received bone marrow from kindlin-2+/− or WT donors. Aortic endothelial cells isolated from WT and kindlin-2+/− mice exhibited similar expression levels of the β1 and β3 integrins as well as VEGFR2. However, FACS analysis of soluble ligand binding showed significantly reduced activation of the β3 integrins in kindlin-2+/− ECs as compared to WT cells, whereas the β1 integrin activation was similar in both genotypes. Additionally, PMA or VEGF enhanced integrin activation in WT ECs, but failed to do so in the kindlin-2+/− cells. Accordingly, integrin-dependent cellular functions: adhesion, migration and spreading of the EC derived from the kindlin-2+/− mice were significantly blunted on the β3 integrin substrates but not on the β1 substrates. In control experiments, overexpression of kindlin-2 in the kindlin-2+/− EC restored defective spreading on vitronectin while a kindlin-2 that does not bind integrins, did not, indicating specificity. Also, tube formation by kindlin-2+/− ECs was significantly impaired compared to those developed by WT ECs. Taken together, kindlin-2 plays an important role in angiogenesis as well as in maintenance of vascular integrity. Disclosures: No relevant conflicts of interest to declare.

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