Location of qPCR amplicon covering region R1 is indicated. -, Semenza GL Targeting HIF-1 for cancer therapy. In endothelial cells (EC), the transcription factor NF-B is important in the switch from quiescence to activation, and is tightly controlled to avoid excessive inflammation and organ damage. Constitutive Wnt signaling activation caused by mutations in -catenin or genes that control -catenin stability has been associated with aberrant cell proliferation and subsequent cancer progression (reviewed in. The endothelium is a single layer of squamous endothelial cells that line the interior surface of blood vessels and lymphatic vessels. (F) Quantification of yolk sac vitelline vessel diameter. Common gene hits and accession numbers from GSEA analysis. Recently we have shown that inhibition of Erg expression using antisense (Genebloc or GB) up-regulates basal ICAM-1 mRNA and protein levels (12). 2020 Aug 1;319(2):H443-H455. Copyright 2022 Elsevier Inc. except certain content provided by third parties. No supershifted band was visible after the addition of the anti NF-B p65 antibody. In agreement with previous data, transduction with AdErg repressed ICAM-1 promoter activity to 38% of treatment with AdLacZ (Fig. Bioinformatic analysis using MatInspector shows that this region contains 16 EBS consensus sequences (Fig. This suggests that ERG may promote stabilization of angiogenesis in part through control of pericyte recruitment. Wnt targets CyclinD1 and Axin2, previously shown to be decreased in ERG-deficient endothelium (see. b: ERG exclusively, Figure 7.. EVI of different CNS lesions,. 3D Reconstruction of Neovessels inside Matrigel Plugs Supplemented with VEGF and Adenovirus Expressing Lacz, Related to Figure6, Movie S2. O-PROTAC offers straight-forward predictability, reprogrammability and superior stability. Disclaimer, National Library of Medicine ERG is a critical regulator of Wnt/LEF1 signaling in prostate cancer. erg (avian v-ets erythroblastosis virus e26 oncogene homolog), member of the ets family transcription factors, is constitutionally expressed in endothelial cells regulating angiogenesis and endothelial apoptosis. (D, panels iii and iv) ChIP was carried out on sheared chromatin from HUVEC treated with control or Erg siRNA, using an anti NF-B p65 or control IgG antibody. Inhibition of Erg in resting HUVEC stimulates leukocyte recruitment (13). Therefore both genes are candidates for regulation by both Erg and NF-B. Background: In quiescent endothelial cells, the transcription factor Erg regulates cell homeostasis by repressing expression of proinflammatory genes. The ERG and FLI1, induces EndMT coupled with dynamic . To investigate whether the same applies to the other target genes investigated here, we tested whether Erg inhibits binding of NF-B p65 to the IL-8 and cIAP2 promoters. Google Scholar. image, Download .pdf (5.91 Other ETS combinatorial transcription factor motifs include the serum response elements that bind ternary complex ETS factors and the serum response factor (45), and ETS:AP1 binding motifs (46). government site. is a recipient of a DOC-fFORTE fellowship of the Austrian Academy of Sciences at the London Research Institute. Takeuchi H, Hashimoto N, Kitai R, Kubota T, Kikuta K. J Neurosurg. HHS Vulnerability Disclosure, Help In conclusion, this set of experiments indicates that Erg-mediated repression of ICAM-1 involves EBS 118 and EBS 181, which is located within the NF-B consensus sequence, and requires a functional NF-B binding site. Bethesda, MD 20894, Web Policies Epub 2014 Oct 28. A recent paper has described a role for ETS factors (including ERG) in arterial specification and reported increased ERG expression in arterial-derived EC invitro (. Red cell distribution width . Oligonucleotides used in this study are described in supplemental Table S1. Typically only the nuclei are visible, at the boundary between the lumen and the wall of a vessel. From 2008 to 2016, a total of $36 946 764.00 USD has . In this study we identify a novel mechanism that controls endothelial quiescence through inhibition of NF-B activity. Endothelial Fli1 deficiency impairs vascular homeostasis: a role in scleroderma vasculopathy. Publishers note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. (B) ERG (magenta), VEC (red), -cat (green), and DAPI (blue) staining of control and ERG-deficient HUVEC transduced with GFP-tagged control (Ctrl-GFP) or VE-cadherin (VEC-GFP) adenovirus. Briefly, HUVEC (3 104 cells/well) seeded onto 1% gelatin-coated 24-well plates in EGM2 were transduced with 50 multiplicity of infection of AdErg or -galactosidase adenovirus (AdLacZ). # Discussion A hemangioma is a usually benign vascular tumor derived from blood vessel cell types. Briefly, cells were incubated with 1.5 l of GeneJuice, 250 ng of pGL4 ICAM-1 1.3, pGL4ICAM1-EBS mutants, or pGL4.10 empty vector and 250 ng of pGL4.73 Rluc/TK Renilla luciferase (Promega) for 24 h. Luciferase activity was measured using the Dual Luciferase Reporter Assay System (Promega) on a Syngergy HT microplate reader (BioTek, Winooski, VT) to calculate the dual luciferase ratio. Taken together, these data suggest that Erg repression involves two EBS, 118 and 181 bp upstream of the transcription start site and either of these sites is sufficient for Erg-mediated repression. Caught up in a Wnt storm: Wnt signaling in cancer. Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. 5C), indicating a trend toward a correlation. Endothelial cells form the barrier between vessels and tissue and control the flow of substances and fluid into and out of a tissue. After 39 h, cells were treated with BAY 11-7085 (5 m, Sigma) diluted in dimethyl sulfoxide and incubated for a further 24 h. Cells were treated with 10 ng/ml of TNF- for the final 6 h. ICAM-1 protein levels were measured by Western blot using an anti-ICAM-1 (clone 15.2, kind gift of Prof. Nancy Hogg) antibody, and normalized to GAPDH (MAB374 Millipore). B.G. In conclusion, the EMSA experiments indicate that Erg binds to both EBS 118 and 181 in the ICAM-1 promoter. 3D Reconstruction of Neovessels inside Matrigel Plugs Supplemented with VEGF and Adenovirus Expressing ERG, Related to Figure6. Chromatin immunoprecipitation (ChIP) was performed using ChIP-IT express (Active Motif, Rixensart, Belgium) as previously described (10). 2C). Accordingly, the expression of ERG in 22Rv1 and C4-2B cells co-cultured with HUVEC was determined, respectively. A. Before 8600 Rockville Pike (D and E) (D) Western blot and (E) qPCR analysis of -catenin expression in control (siCtrl) and ERG-deficient (siERG) HUVEC (n= 4). Moreover we showed that overexpression of Erg inhibited acute inflammation in mouse paws induced by TNF- (12). Interestingly, ERG overexpression in the invivo Matrigel plug model resulted in increased pericyte recruitment to vessels. 1 The transcriptional mechanisms regulating EC lineage identity and maintenance of endothelial homeostasis are also areas of immense interest for vascular regenerative therapies but remain poorly understood. Members of the transcription factor family nuclear factor (NF)-B are important mediators of proinflammatory responses in the vasculature. a: H & E demonstrates pseudopalisading cells surrounding areas of central necrosis,, Figure 3.. GBM, glomeruloid type. 2010 Jun 8;121(22):2407-18. doi: 10.1161/CIRCULATIONAHA.110.938217. 3B), in fact p65 does not bind to this same sequence in native chromatin from unstimulated HUVEC. Intriguingly, ERG knockdown in human umbilical vein endothelial cells (HUVECs) promoted the secretion of endothelin-1 (ET-1), which subsequently accelerated the proliferation, phenotypic transition, and collagen synthesis of cardiac fibroblasts in a paracrine manner. Molecular regulation of vessel maturation. DNA protein interactions were then investigated using the Lightshift chemiluminescent EMSA kit (Pierce). We have previously investigated the genome-wide targets of Erg by transcription profiling of HUVEC treated with Erg Genebloc (16), and found that Erg represses a number of genes involved in inflammatory processes. 2021 Sep;21(9):2950-2963. doi: 10.1111/ajt.16488. HUVEC were seeded onto 1% gelatin-coated plates and grown in EGM-2 medium (Lonza, Wokingham, United Kingdom). ENCODE ChIP-seq data profiles for H3K4me1, H3K27Ac, and RNA polymerase II indicate open chromatin and active transcription. Shifted protein-oligonucleotide complexes are indicated by an arrow and super-shifted complexes are indicated by arrowhead. Images are a single representation of at least 3 separate experiments. ERG can serve as a reliable marker of immature myeloid cells in cases of extramedullary hematopoiesis, bone marrow trephine biopsies and in adrenal myelolipomas ( Am J Surg Pathol 2011;35:432, Hum Pathol 2022;124:1 ) Reliably identifies the presence of ERG fusions in Ewing sarcomas ( Mod Pathol 2012;25:1378, Genes Chromosomes Cancer 2016;55:340 ) It is possible that some of the genes may be indirect targets of Erg; genomic ChIP sequencing of Erg DNA binding profiles will address this question. HHS Vulnerability Disclosure, Help In the latest study, researchers found that loss of function of this ERG master gene in endothelial cells caused damage and fibrosis in the livers of mice. Immunoprecipitated DNA was analyzed by qPCR for primers covering ICAM-1 promoter regions (R) 35 (C) or 4 (D) and negative control region. Although EndMT is observed in various diseases including cancer, and augments fibrosis and vascular defects, the mechanism of EndMT induction is not fully understood. Endothelial cells were selected by magnetic separation and plated in primary 96-well plates coated with fibronectin/gelatin in EGM-2 medium. Collectively, endothelial ERG alleviates cardiac fibrosis via blocking ET-1-dependent paracrine mechanism and it functions as a candidate for treating cardiac fibrosis. 2 erg has been recently studied immunohisto-chemically in prostate carcinoma, subsets of which have oncogenic tmprss2-erg gene fusions Immunoprecipitated DNA was analyzed by qPCR for primers covering ICAM-1 promoter regions 15 (D) or region 4 only (E) and negative control region. Thought to be due to vascular endothelial growth factor; . NF-B activity was inhibited using an adenovirus expressing a mutant super repressor version of IB (AdIBSR), which cannot be phosphorylated and degraded, resulting in sequestration of NF-B in the cytoplasm (14). Erg represses ICAM-1 and binds to the ICAM-1 promoter region 4 in resting EC. We thank Dr. Odile Dumont, Dr. Richard Starke, Professor Guido Franzoso, Dr. Paul Evans, Professor Malcolm Parker, and Dr. Irina Udalova (Imperial College London) for discussions. ETS-related gene (ERG) is a transcription factor that has been linked to angiogenesis. This is likely to be the result of endothelial activation by proinflammatory stimuli, because Erg levels have been shown to decrease upon LPS or TNF- stimulation (9, 13). Very little research has been done to assess ERG expression in central nervous system (CNS) tumors. ChIP was carried out on a confluent monolayer of quiescent or TNF--treated HUVEC. The cells can This subclass of tumour cells shows low AR expres- then remain dormant, interacting with native cells in sion and concomitant reduced PSA expression, which the niche, before proliferating to form a new tumour, frequently occurs in combination with loss of both which in turn has the . (G and H) Panels show endomucin staining of blood vessels in B16F0 tumors and the quantification of the number of endomucin-positive vessels, (n= 6), scale bar, 50m. The emerging role of leptin in obesity-associated cardiac fibrosis: evidence and mechanism. Comparison between the Erg dataset and the genes up-regulated in pancreatic cancer cells after TNF- treatment gave a significant normalized enrichment score of 1.67 (Fig. Immunoprecipitated DNA was analyzed by qPCR for primers covering the NF-B site and EBS in cIAP2 (D, panel iii), or IL-8 (panel iv) (see supplemental Fig. Nat Commun. Previously, using an ICAM-1 promoter luciferase construct containing the first 1.3 kb upstream of the ICAM-1 transcription start site, we have shown that Erg represses ICAM-1 promoter activity in resting EC (12). Under hyperglycemic conditions, the endothelial cells become dysfunctional. From the National Heart and Lung Institute (NHLI) Cardiovascular Sciences Unit, Hammersmith Hospital, Imperial College London, London W12 0NN and, the Department of Haematology, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, United Kingdom. Comparative genomic analysis of the Fzd4 promoter revealed the presence of three highly conserved ERG DNA binding motifs in the 800 base pair (bp) region upstream of the Fzd4 transcription start site (, Wnt/-catenin signaling can promote EC proliferation (. In endothelial cells (EC), the transcription factor NF-B is important in the switch from quiescence to activation, and is tightly controlled to avoid excessive inflammation and organ damage. (1996), Regulated expression of the ets-1 transcription factor in vascular smooth muscle cells, Cheng C., Tempel D., Den Dekker W. K., Haasdijk R., Chrifi I., Bos F. L., Wagtmans K., van de Kamp E. H., Blonden L., Biessen E. A., Moll F., Pasterkamp G., Serruys P. W., Schulte-Merker S., Duckers H. J. 176, 19831998 (2010). Retinas were collected at P6 and processed as described (. S3) and a negative control GAPDH promoter region. The content on this site is intended for healthcare professionals. -, Hardee ME Zagzag D Mechanisms of glioma-associated neovascularization. Dynamic regulation of canonical TGFbeta signalling by endothelial transcription factor ERG protects from liver fibrogenesis. Erg expression was inhibited using either Genebloc (Silence Therapeutics AG, Berlin, Germany) as previously described (10), or by RNA interference with short interfering RNA (siRNA). Mutations within more than one EBS were carried out sequentially. Focusing on intercellular adhesion molecule 1(ICAM)-1 as a model, we identify two ETS binding sites (EBS 118 and 181) within the ICAM-1 promoter required for Erg-mediated repression. Lithium inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells. However, loss of the major complex after addition of NF-B p65 antibody suggests that, at least in vitro, nuclear NF-B p65 is able to bind to the EBS 181 oligonucleotide in resting HUVEC as well as in activated HUVEC. . is a recipient of a National Lung and Heart Institute Foundation Studentship. He was discharged from our hospital on postoperative day 6. Common genes enriched for Erg GSEA among all three studies are in dark grey. To expand this analysis to the entire Erg dataset and find patterns that could support the biological data, we used GSEA (3638), and compared the Erg dataset with other relevant gene sets, to identify common biological functions. The above data suggest that Erg is repressing ICAM-1 through a mechanism that involves EBS 118 and EBS 181. However, a combination of ETS factors ETV2, FLI1, and ERG is a potent endothelial inducer from human amniotic cells . Acute myeloid leukemia with complex karyotypes and abnormal chromosome 21: amplification discloses overexpression of APP, ETS2, and ERG genes. (A) Staining for VE-cadherin (green), ERG (red), and isolectin B4 (IB4, blue) in, (B) Relative mRNA expression of Erg and VE-cadherin in primary. and transmitted securely. Our previous studies have shown that ERG expression is highly enriched in endothelial cells (EC) both in vitro and in vivo. 2015 Jan;68(1):44-50. doi: 10.1136/jclinpath-2014-202629. Accessibility Because EMSA studies have shown that Erg can bind to this site, we investigated the possibility that, in resting EC, Erg might block NF-B p65 binding to the ICAM-1 promoter at EBS 181. PMC endothelial junctions are crucial for the maintenance and regulation of vascular homeostasis and function and mediate a complex signaling network. Epub 2010 May 24. (2021). Xu P, Ge FH, Li WX, Xu Z, Wang XL, Shen JL, Xu AB, Hao RR. Circ. Analysis of the three studies together suggests a strong correlation, although not complete overlap, between genes repressed by Erg and genes transactivated by NF-B. Endothelial-to-mesenchymal transition (EndMT) is a phenomenon in which endothelial cells lose their characteristics and acquire mesenchymal-like properties. A, Erg siRNA or control siRNA-treated HUVEC were transduced with AdIBSR or AdLacZ. These results identify a role for Erg as a gatekeeper controlling vascular inflammation, thus providing an important barrier to protect against inappropriate endothelial activation. A number of these EBS have been shown to play a role in ICAM-1 expression, including two AP1-EBS repeats involved in ICAM-1 induction after H2O2 stimulation (24), and two EBS that are involved in ICAM-1 expression in non-endothelial cells (2527). 1A). Because Erg also binds to this region, we investigated whether the presence of Erg inhibits the binding of NF-B p65 to DNA, by performing ChIP for NF-B p65 on HUVEC treated with Erg siRNA. Common genes enriched for Erg GSEA with TNF--treated pancreatic cancer cells and TNF--treated EC studies are in light grey. Differentiated chondrocytes showing stem cell markers indicate the transit-amplifying nature of Studies have also demonstrated that the synovium these cells but their significance in cartilage regen-and infrapatellar fat pad contain mesenchymal eration is still unclear [Blanpain et al. 3B, lane 1, arrows), which were competed by excess unlabeled probe but not by a probe containing a mutation in the EBS 181 site (Fig. Claudin 4 identifies a wide spectrum of epithelial neoplasms and represents a very useful marker for carcinoma versus mesothelioma diagnosis in pleural and peritoneal biopsies and effusions There are various types of hemangiomas. Erg may similarly repress NF-B p65 binding by recruiting co-repressors to the promoters of NF-B target genes. More recently, ETV2 has been reported to transdifferentiate skeletal muscle into functional ECs in zebrafish . Healthy EC maintain homeostasis through a dynamic balance between expression of protective genes and repression of proinflammatory genes. (2006), The principal eosinophil peroxidase product, HOSCN, is a uniquely potent phagocyte oxidant inducer of endothelial cell tissue factor activity. Conserved and evolved features, Yang L., Xia L., Wu D. Y., Wang H., Chansky H. A., Schubach W. H., Hickstein D. D., Zhang Y. We suggest that in quiescent EC, Erg acts as a gatekeeper, to inhibit the activity of low constitutive levels of nuclear NF-B p65 and protect the endothelium from the up-regulation of proinflammatory genes, thus maintaining quiescence. (I) TCF reporter (TOP) activity in control and ERG-deficient HUVEC treated with rWnt3a. Dev. The double mutant EBS 96/118, shown previously to have a role in both transactivation and repression of ICAM-1 by ETS factors in rabbit kidney RK13 cells and HEK 293 cells (2527), also showed no increase in promoter activity after Erg inhibition; however, this is likely to occur through EBS 118, as mutation of EBS 96 alone was responsive to Erg inhibition. One of these sites (EBS 181) is located within the consensus binding site for NF-B. 2001;161:III-XIII, 1-151. doi: 10.1007/978-3-642-56553-3. Finally, mutation of the NF-B site at 188 resulted in loss of ICAM-1 up-regulation following Erg Genebloc, indicating that NF-B is involved in the Erg-dependent repression. Mol Cell Biochem. 2000; 88: 26062618. Regulation of expression and activation by interleukin-1 and tumor necrosis factor , Rudders S., Gaspar J., Madore R., Voland C., Grall F., Patel A., Pellacani A., Perrella M. A., Libermann T. A., Oettgen P. (2001), ESE-1 is a novel transcriptional mediator of inflammation that interacts with NF-B to regulate the inducible nitric-oxide synthase gene, Grall F., Gu X., Tan L., Cho J. Y., Inan M. S., Pettit A. R., Thamrongsak U., Choy B. K., Manning C., Akbarali Y., Zerbini L., Rudders S., Goldring S. R., Gravallese E. M., Oettgen P., Goldring M. B., Libermann T. A. Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. Dev. This article contains supplemental Table S1 and Figs. Cooperation of liver cells in health and disease. 2014, Received in revised form: 2017;8(1):895. To confirm the specificity for Erg binding at this site, ChIP was carried out on chromatin from HUVEC treated with Erg siRNA. Embryonic Pericytes Promote Microglial Homeostasis and Their Effects on Neural Progenitors in the Developing Cerebral Cortex. The morphology of the LNCaP-androgen independent (AI) cell line was observed. The ability of BAY-117085 to repress the NF-B pathway was confirmed in TNF- treated HUVEC (supplemental Fig. S1 and Ref. Data are expressed as fold-change compared with IgG normalized to input and control region (n = 8). government site. In this study, we use genetic lineage-specific mouse models and multiple invitro models to show that ERG promotes vascular growth and stability, through control of the canonical Wnt/-catenin pathway. Federal government websites often end in .gov or .mil. Am J Pathol. HUVEC were isolated and cultured in supplemented M199 media as previously described (10). Cooperative ETS transcription factors enforce adult endothelial cell fate and cardiovascular homeostasis. ERG; Endothelial cells; Endothelin-1; Fibroblasts; Paracrine. These data show that the endothelial TF ERG is essential to protect from spontaneous thrombosis in selected vascular beds. 5D, panel ii, and supplemental Fig. Because Erg inhibits ICAM-1 expression by repressing NF-B activity, we hypothesized that Erg may also inhibit the expression of other NF-B target genes in resting endothelium. ERG is abundant in murine hearts, especially in cardiac ECs, but decreased during fibrotic remodeling. EBS were mutated from GGAA to CCAA. official website and that any information you provide is encrypted In this study we investigate the mechanisms used by Erg to repress inflammatory gene expression in quiescent EC, focusing on ICAM-1 as a model gene. All graphical data are SEM, Wnt ligands bind to receptors of the Fzd family to inhibit the -catenin degradation complex and activate Wnt signaling (. The 3D invitro model of angiogenesis was performed as described previously (. This study aims at investigating the potential role and molecular basis of ERG in fibrotic remodeling within the adult heart. 2022 Jul 25;13(1):4170. doi: 10.1038/s41467-022-31890-4. 2022 Feb;102(2):204-211. doi: 10.1038/s41374-021-00698-z. Chromatin was immunoprecipitated with 2 g antibody to Erg (sc-353, Santa Cruz Biotechnology, Inc), NF-B p65 (ab7970, AbCam, Cambridge, United Kingdom), or negative control rabbit IgG (PP64, Chemicon, Millipore). The site is secure. ERG silence in HUVECs promotes the secretion of endothelin-1, which in turn activates cardiac fibroblasts in a paracrine manner. To confirm that ERG controls angiogenesis and vascular development in a Wnt/-catenin-dependent manner invivo, we carried out a rescue experiment by pharmacological stabilization of Wnt/-catenin signaling (. Bookshelf granulocyte-macrophage colony-stimulating factor (gm-csf) is produced in diverse cns cells under pathological conditions, 18 - 20 and it can cross the bbb. n = 4 (C), n = 6 (D); *, p < 0.05. S1S3. https://doi.org/10.1038/s44161-022-00144-3. However, Erg overexpression repressed the EBS 118 and 181 constructs when these EBS were mutated individually (Fig. MeSH An official website of the United States government. Suppressing ET-1 with either a neutralizing antibody or a receptor blocker abolished ERG knockdown-mediated deleterious effect in vivo and in vitro. B, ICAM-1 promoter activity of EBS mutants after Erg Genebloc treatment. In resting endothelial cells (EC),2 NF-B is sequestered in the cytoplasm by proteins of the inhibitor of NF-B (IB) family. The ICAM-1 promoter EBS mutants 96, 153, 358, 834, and 907 also showed significantly increased promoter activity after Erg inhibition. The above data suggest that Erg is repressing ICAM-1 expression by inhibiting NF-B p65 binding to the ICAM-1 promoter. C and D, ChIP was carried out on sheared chromatin from confluent resting HUVEC TNF (10 ng/ml for 30 min) (C), or HUVEC treated with Erg or control siRNA (D) using an anti-NF-B p65 or control IgG antibody. Therefore we speculated that there may be an inhibitory element in the native chromatin structure of the ICAM-1 promoter that inhibits the binding of p65 to this site. 5D, panels iii and iv, respectively). a: Original image of a GBM stained with, Figure 2.. GBM. To obtain These exceptions notwithstanding, ERG is the most sensitive and specific endothelial marker available. Nrarp coordinates endothelial Notch and Wnt signaling to control vessel density in angiogenesis. Erg is a member of the ETS family of transcription factors, characterized by a conserved ETS DNA binding domain that binds to a core consensus motif of GGA(A/T) (6). Using the inducible endothelial specific Cdh5(PAC)-iCreERT2 line, we show that ERG is required for angiogenesis in the developing retina of newborn mice and for tumor blood vessel growth in adult mice. and S.T.K. Res. 2003; 3: 721732. Therefore Erg represses expression of multiple NF-B target genes in resting EC. These results indicate that, as with ICAM-1, Erg inhibits constitutive binding of nuclear NF-B p65 to the promoters of IL-8 and cIAP2. 2022 Oct 10. doi: 10.1007/s11010-022-04562-6. Smad-dependent and Smad-independent pathways in TGF-beta family signalling. MicroRNA-147a Targets SLC40A1 to Induce Ferroptosis in Human Glioblastoma. (H) mRNA expression of Erg, -catenin, and its target genes Cyclin D1, Axin-2, and TCF-1 in primary, (I) qPCR analysis of total brain mRNA from control and, (J) GSEA shows enrichment and significant correlation (normalized enrichment score, 2.46; p< 0.001) between genes downregulated in -catenin siRNA-treated HPAEC (green curve) (. Unable to load your collection due to an error, Unable to load your delegates due to an error. Addition of IgG antibodies had no effect on the band pattern (Fig. This consensus sequence appears in the ICAM-1 promoter five times, at 1239, 1136, 773, 118, and 75 relative to the transcription start site (Fig. Functionally, knockdown of ERG and FLI1 induced cell monolayer permeability with a potency similar to that of vascular endothelial growth factor. The graphical outputs show enrichment (green curve) of genes up-regulated in TNF--treated pancreatic cancer cells (A), endothelial cells (B), or genes bound by NF-B p65 in macrophages after LPS stimulation (C), along a ranked list of genes up- or down-regulated by 48 h of Erg inhibition. The ICAM-1 promoter construct pGL4 ICAM-1 1.3 containing the first 1.3 kb upstream from the transcription start site as described in Ref. J Pers Med. (BD) (B) Representative images of EC sprouts on fibrin gel beads using siCtrl or siERG-treated HUVEC in the presence or absence of LiCl; (C) quantification of numbers of sprouts; and (D) tube length (n= 20). 2022 Dec;58:102513. doi: 10.1016/j.redox.2022.102513. Immunohistochemistry analysis revealed endothelial cells stained positive for ERG, D2-40 (podoplanin) and negative for WT1, calretinin. The chromatin-remodeling enzyme BRG1 modulates vascular Wnt signaling at two levels. Endothelial Cells. Dynamic Phosphorylation of CENP-A at Ser68 Orchestrates Its Cell-Cycle-Dependent Deposition at Centromeres, Reverse Genetic Screening Reveals Poor Correlation between Morpholino-Induced and Mutant Phenotypes in Zebrafish, Endothelial ERG Is Required for Vascular Development, Angiogenesis, and Tumor Growth, ERG Is Required for Vascular Development, Physiological Postnatal Angiogenesis, and Pathological Tumor Angiogenesis, ERG Controls Vascular Stability and Pericyte Coverage, ERG Controls -Catenin Stability and Signaling through VE-Cadherin- and Wnt-Dependent Mechanisms, Endothelial Canonical Wnt Signaling and -Catenin Stability Are Regulated by ERG, ERG Controls -Catenin Stability through VE-Cadherin- and Wnt-Dependent Mechanisms, Expression of the Wnt Receptor Frizzled-4 Is Regulated byERG, Baylor College of Medicine Human Genome Sequencing Center, Washington University Genome Sequencing Center, Childrens Hospital Oakland Research Institute, ERG Controls Angiogenesis through Wnt Signaling, ERG Regulates Angiogenesis through Wnt/-Catenin Signaling, ERG Overexpression Stabilizes VEGF-Induced Blood Vessels and Promotes Angiogenesis InVivo, Isolation of Mouse Lung Endothelial Cells, Plasmid Transfections and Reporter Assays, eyJraWQiOiI4ZjUxYWNhY2IzYjhiNjNlNzFlYmIzYWFmYTU5NmZmYyIsImFsZyI6IlJTMjU2In0.eyJzdWIiOiI0MTE4MjEwN2VhMzAwMjI5Y2M1YTE0ZWY1ZDllNjA4OCIsImtpZCI6IjhmNTFhY2FjYjNiOGI2M2U3MWViYjNhYWZhNTk2ZmZjIiwiZXhwIjoxNjcwODIyNzE3fQ.n2t_EEIadOWDuBvYBM3qxbgnXUkqeUfJTf3R687qKZj_pnMqh3eIgs7tPeGSFW6wtKoPQsvfeQ2e5TaMM76aisnz580wH5QVG1dLFDmHCQpgnnDOct2_VHnSBX1CGX83q7xhXMGqUOUE-HNIfQxZ53P5GwfcnHh1lLJU74cT2FkyUg4J_mvbMyU9e2FhBmwbjsBfMQeAEIM0gKLuvF4lAhAd_IXnZlTR5ly4rlairqiIeoKF1qr6Q53jhqOvogcfAuJtwEenI0cy2VsXzNimB5zlXFGUZGkjHhhrf0_KiHdfZfw9C0ffYD0fg93UcxK4QXJzlX7g01j7zDxB91xtIw, eyJraWQiOiI4ZjUxYWNhY2IzYjhiNjNlNzFlYmIzYWFmYTU5NmZmYyIsImFsZyI6IlJTMjU2In0.eyJzdWIiOiI4ZjMzNWYwNjIyYjJmYmIzZmM0Y2M4ZGYwYzY5MzJmNCIsImtpZCI6IjhmNTFhY2FjYjNiOGI2M2U3MWViYjNhYWZhNTk2ZmZjIiwiZXhwIjoxNjcwODIyNzE3fQ.idar2V0x9cfoVI0p0kPMyjqj1i5F1H25HkaKd9fNUWIKAtgAtdUoD6q69na4nkTzLP-H2cJzVfx2bpwJOyx-S1IpEqKf0-YyNA0Nea9nSSHrbqGDpCi6GHWiUSM-V27KBzit9SsbsWal6a1aQMSbDQL1S_Y9POeP40CfrC_s4Pe6PAz-Gagozm5_MCh4FenMF6DVpiL0Ts3Vvz85NVmt0how8309XLFsJL_IrMuonIQjU6L71zYGkwTTdoU4D1JNyfLkSAxnoKAAPNkfZM61-fW0sLo7omTLvNNhAJQS7kv7ofGuPMeCa6yNbR7Q6Qa4ENhb7HQuQTRdq5xZKyhFew, https://doi.org/10.1016/j.devcel.2014.11.016, The Endothelial Transcription Factor ERG Promotes Vascular Stability and Growth through Wnt/-Catenin Signaling, View Large Dapper 1 antagonizes Wnt signaling by promoting dishevelled degradation. the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in FEV contains an ETS DNA binding domain, which is highly homologous to Erg and Fli-1 (48); however, its restricted pattern of expression suggests that FEV is not likely to play a role in transcriptional repression in endothelial cells. The more common types include capillary, cavernous, combined, and lobular capillary types. Epub 2021 Feb 2. Unable to load your collection due to an error, Unable to load your delegates due to an error. A.V.S. 3B, lane 11 compared with lane 1). To identify which EBS are involved in Erg-mediated repression, we generated ICAM-1 promoter constructs with mutations within single EBS, or two EBS together, if they had previously been shown to have a cooperative role (24, 25). ERG and FLI1 protein expression in epithelioid sarcoma. 2008;111(7):3498506. Acta Neuropathol. In combination with the current therapy, SSA had potential to improve treatment effectiveness and to prevent cancer recurrence. Mouse melanoma B16F0 tumors were grown in tamoxifen-treated adult. Endothelial cell (EC) plasticity in pathological settings has recently been recognized as a driver of disease progression. Before Vascular morphogenesis: a Wnt for every vessel?. S3). ERG deficiency results in vessel regression and reduced pericyte recruitment, demonstrating that ERG controls vascular stability. E26 transformation-specific (ETS)-related gene (ERG) belongs to the ETS transcriptional factor family and is required for endothelial cells (ECs) homeostasis and cardiac . Numerous studies have shown that the new vasculature induced by VEGF invivo, to promote revascularization in ischemic diseases (. (F) Quantification of pericyte coverage, pixel intensity (n= 8). More importantly, we proved that endothelial ERG overexpression notably prevented pressure overload-induced cardiac fibrosis. Accepted: Endothelial Forkhead Box Transcription Factor P1 Regulates Pathological Cardiac Remodeling Through Transforming Growth Factor-1-Endothelin-1 Signal Pathway. 2022 Sep 16;23(18):10848. doi: 10.3390/ijms231810848. 3A, solid arrow), indicating that Erg binds to the EBS 118 site. 1C). Federal government websites often end in .gov or .mil. B-cell lymphoblastoid cell lines, encoding 16 different DPB1 alleles, were studied. Interestingly, inhibition of Erg expression in quiescent EC results in increased NF-B-dependent ICAM-1 expression, indicating that Erg represses basal NF-B activity. Here, we applied CoBATCH for single-cell epigenomic tracing of dynamic EC lineage histories in five mouse organs from development to ageing. -, Louis DN Ohgaki H Wiestler OD Cavenee WK Burger PC Jouvet A Scheithauer BW Kleihues P The 2007 WHO classification of tumours of the central nervous system. The ETS transcription factor family is implicated in vascular development and angiogenesis (reviewed in. PLOS Genetics, 14 (11), e1007826 | 10.1371/journal.pgen.1007826 sci hub to open science save Nagai, N., Ohguchi, H., Nakaki, R., Matsumura, Y., Kanki, Y., Sakai, J., Minami, T. (2018). Co-staining for isolectin B4 and the basement matrix component collagen IV showed a greater number of empty collagen IV sleeves in the capillary plexus (, (A) Collagen IV (green) and isolectin B4 (IB4, red) staining of, (B) NG2-positive pericytes (green) associated with isolectin B4 labeled retinal vessels (red) from, (C) Sections from B16F0 tumors grown on adult, Invitro studies have shown that ERG is essential to maintain theintegrity of endothelial junctions, by driving expression of VE-cadherin (. S3) and negative control GAPDH promoter region. Of the top 9 studies with the most significant overlaps in regulated genes, two involved the response to treatment with the NF-B activating cytokine TNF-. Cardiovas. Image, Download Hi-res This, in conjunction with our present study of the basal repression of proinflammatory genes, suggests that inhibition of Erg is required to induce a NF-B-mediated inflammatory response in the endothelium, and that Erg can control NF-B activity at multiple levels. We show that constitutive endothelial deletion of ERG (, Angiogenesis is essential during embryogenesis and is a critical component of many diseases. We show that in quiescent EC Erg prevents NF-B p65 binding to DNA, suggesting that Erg may compete with p65 for DNA binding. Endothelial cell-derived non-canonical Wnt ligands control vascular pruning in angiogenesis. We propose that Erg may provide a checkpoint to pass before induction of NF-B target genes occurs in EC. It has been recently shown that the ETS factor ERG interacts with endothelial-specific genes, including vWF . (B) Vessel permeability was quantified by measuring the amount of dextran-TRITC present outside of the dextran-FITC positive vessels, arbitrary units (n= 3). Epub 2022 Oct 22. The transcription factor Erg inhibits vascular inflammation by repressing NF-kappaB activation and proinflammatory gene expression in endothelial cells. Endothelial cytoplasm is inconspicuous in routine light microscopy. Cell 32, 82-96 (2015). Proliferation of vascular smooth muscle cells in glioblastoma multiforme. Please enter a term before submitting your search. Stockman DL, Hornick JL, Deavers MT, Lev DC, Lazar AJ, Wang WL. Cultured endothelial cells display endogenous activation of the canonical Wnt signaling pathway and express multiple ligands, receptors, and secreted modulators of Wnt signaling. Statistical significance was determined by using unpaired two-tailed Students t test. This smooth anticoagulant surface functions as a selective filter to regulate the passage of gases, fluid, immune cells, and various molecules. (D and E) (D) qPCR and (E) western blot analysis of Fzd4 expression in control and ERG-deficient cells (n= 3). In this study, we investigated the miRNA expression changes . Overlapping and divergent signaling pathways of N-cadherin and VE-cadherin in endothelial cells. ETS factors regulate specific target genes through combinatorial promoter motifs and interaction with other transcription factors. The endothelial transcription factor ERG promotes vascular stability and growth through Wnt/-catenin signaling. Federal government websites often end in .gov or .mil. Previous studies have demonstrated that Erg binds to an EBS in the IL-8 promoter (13). Chen R, Cao C, Liu H, Jiang W, Pan R, He H, Ding K, Meng Q. Redox Biol. Hypoxia is implicated in loss of retinal ganglion cells (RGCs) occurring in such conditions. Nature. In support of the specificity of the interaction of Erg with this site, addition of control IgG antibodies had no effect on the band pattern (Fig. The https:// ensures that you are connecting to the (1997), A new member of the ETS family fused to EWS in Ewing tumors, Carrre S., Verger A., Flourens A., Stehelin D., Duterque-Coquillaud M. (1998), Erg proteins, transcription factors of the Ets family, form homo-, heterodimers, and ternary complexes via two distinct domains, Gottschalk L. R., Giannola D. M., Emerson S. G. (1993), Molecular regulation of the human IL-3 gene. Luscinskas (Harvard Medical School, Boston, MA) for providing the VE-cadherin GFP adenovirus; L. Lawrence (Imperial College London, UK) for technical assistance; G. Cossu (University of Manchester, UK) for critical reading of the manuscript; and A. Taddei (Cancer Research UK, London, UK), M. Johns (Imperial College London, UK), and D. Haskard (Imperial College London, UK) for helpful discussions. (2010), Genome-wide analysis of ETS family DNA-binding, Wilson N. K., Foster S. D., Wang X., Knezevic K., Schtte J., Kaimakis P., Chilarska P. M., Kinston S., Ouwehand W. H., Dzierzak E., Pimanda J. E., de Bruijn M. F., Gttgens B. (2002), Molecular cloning of ESET, a novel histone H3-specific methyltransferase that interacts with ERG transcription factor, Yang L., Mei Q., Zielinska-Kwiatkowska A., Matsui Y., Blackburn M. L., Benedetti D., Krumm A. B., Tiruppathi C. (2009), NF-B regulates thrombin-induced ICAM-1 gene expression in cooperation with NFAT by binding to the intronic NF-B site in the ICAM-1 gene, Pierce J. W., Schoenleber R., Jesmok G., Best J., Moore S. A., Collins T., Gerritsen M. E. (1997), Novel inhibitors of cytokine-induced IB phosphorylation and endothelial cell adhesion molecule expression show anti-inflammatory effects, Subramanian A., Tamayo P., Mootha V. K., Mukherjee S., Ebert B. L., Gillette M. A., Paulovich A., Pomeroy S. L., Golub T. R., Lander E. S., Mesirov J. P. (2005), Gene set enrichment analysis. Article To investigate whether Erg binding to the ICAM-1 promoter is localized around these sites, ChIP was carried out by scanning the EBS in the first 1.3 kb of the ICAM-1 promoter, using quantitative PCR (qPCR) with primers for regions (R) named 1, 2, 3, 4, and 5 (Fig. will also be available for a limited time. Am. 2004;101(11):391520. Canonical Wnt signaling promotes EC survival, junction stabilization, proliferation, and pericyte recruitment and is essential for vessel stability (. The new PMC design is here! Tight control of NF-B activation is crucial to cellular homeostasis, and several mechanisms, both at the transcriptional and non-transcriptional levels, have been identified. Get the most important science stories of the day, free in your inbox. Additionally, we mutated the NF-B site at 188, previously identified as important for cytokine-mediated up-regulation of ICAM-1 (15) (Fig. Bethesda, MD 20894, Web Policies This is a preview of subscription content, access via your institution, Get immediate online access to Nature and 55 other Nature journal. 2005 Mar;38(1):36-42. doi: 10.1007/s00795-004-0273-0. The remaining genes were commonly enriched between genes bound by NF-B p65 after LPS treatment, and either TNF--treated pancreatic cancer cells or TNF--treated EC. We first tested whether NF-B p65 could bind to the region containing the NF-B site and EBS 181 within the native chromatin structure of resting HUVEC, using ChIP. 2B). A role for the beta-catenin/T-cell factor signaling cascade in vascular remodeling. performed bioinformatic analysis and interpretation; and L.R. Finally, in this study, we explore the potential for ERG in promoting vascular stability during VEGF-induced angiogenesis. RGC death occurs by apoptosis or necrosis. Supplemental Experimental Procedures and Figures S1S6, Movie S1. Although the majority of NF-B p65 in resting EC is located in the cytoplasm, low levels of p65 are also found in the nucleus (12, 31, 32). The inhibitory activity of AdIBSR on NF-B was confirmed on TNF--stimulated HUVEC (supplemental Fig. D, panel ii, ChIP was carried out on sheared chromatin from confluent resting HUVEC, using an anti-Erg or control IgG antibody. (1990), The ETS domain. Using EMSA, we investigated whether, in resting HUVEC, p65 interacts with the NF-B site at 188, which overlaps with EBS 181. All graphical data are SEM, To investigate the role of ERG in physiological and pathological postnatal angiogenesis, floxed. Loss of ERG expression is associated with diseases including atherosclerosis. Wnt/beta-catenin/Tcf signaling induces the transcription of Axin2, a negative regulator of the signaling pathway. We show that Erg binds to both EBS 118 and EBS 181, the latter located within the NF-B binding site. (F) Representative images of B16F0 tumors which were grown for 14days on adult. There was no recurrence at the 1-year follow up. Results are expressed as fold-change compared with IgG normalized to input and negative control region. The enhancers of endothelial specific genes are synergistically activated by FOX and ETS transcription factors containing FOXO:ETS motifs (44). The ETS transcription factor ERG drives expression of VE-cadherinand controls junctional integrity. 3C). Overlapping ETS and NF-B binding sites have been identified in the regulatory regions of inflammatory genes such as IL-3, IL-12, IL-2, IL-2 receptor-, and granulocyte-macrophage colony stimulating factor (4952). siRNA to Fli-1 inhibited expression of Fli-1 but not Erg, Ets-2, or GAPB; similarly, siRNA to Ets-2 affected only Ets-2 expression, not Erg, Fli-1, or GAPB expression; GABP- siRNA only affected GABP- expression and not Erg, Fli-1, or Ets-2; and finally Erg siRNA did not affect expression of the other three ETS factors (Fig. 2015 The Authors. Circulation. Cancer. Specificity was measured by addition of saturating amounts of competing oligonucleotide (competitor) or competing oligonucleotide with a mutation in the EBS 118 (A) or 181 (B) (M-competitor). provided advice on retina isolation and optimized ERG retinal staining; E.D. Alternatively, after 42 h following adenovirus transduction, cells were treated with 10 ng/ml of TNF- for 6 h. ICAM-1 mRNA levels were assessed by quantitative RT-PCR, normalized to GAPDH. 7): Fli-1, the ETS factor with the closest homology to Erg; Ets-2, a more distantly related ETS factor known to interact with Erg (20); and GABP, which can act as an activator or repressor of transcription (21, 22). Am J Surg Pathol. Professor Nancy Hogg (Cancer Research UK) for the kind gift of the anti-ICAM-1 antibody, clone 15.5. HES6, C-HAIRY1, HES-6, bHLHb41, bHLHc23, hes family bHLH transcription factor 6. Hyperglycemia increases the risk of vascular complications such as diabetic retinopathy, diabetic nephropathy, peripheral vascular disease and cerebro/cardiovascular diseases. Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project. EBS 834 and 907, suggested to have a role in H2O2-mediated activation of the ICAM-1 promoter (24), do not appear to have a role in Erg-mediated repression. Whether Erg represses NF-B target genes through recruiting these or other co-repressors is being investigated. These observations are in line with a previous report where global deletion of a subset of endothelial ERG isoforms resulted in vascular defects and lethality between E10.5 and E11.5 (. (A) 3D rendering of confocal microscopy images of whole-mount Matrigel plugs perfused with the dextran tracers. Before Analysis of the IL-8 promoter showed that the EBS identified as the binding site of Erg is located within the functional NF-B binding site, similarly to what was observed for ICAM-1 and cIAP2 (supplemental Fig. b: ERG, Figure 5.. HB. b: ERG exclusively, Figure 6.. Metastatic carcinoma. Ets-1, Ets-2, and ERM overexpression in RK13 cells increased ICAM-1 promoter activity through EBS 118 (25). Combined genomic and antisense analysis reveals that the transcription factor Erg is implicated in endothelial cell differentiation. Gene set enrichment analysis (GSEA) overlap studies were carried out using GSEA software version 2.0 (Broad Institute). INTRODUCTION HISTORY 2003;425(6958):57784. *, p < 0.05; **, p < 0.01; ***, p < 0.001. 4B). FEV expression is restricted to prostate and small intestine tissue (47) and Dami megakaryocytic cells (27). Androgen receptor levels identification were detected in qRT-PCR and Western blot assay. VEGF has been shown to induce the formation of unstable and highly permeable vessels invivo (. eCollection 2022. Baldus CD, Liyanarachchi S, Mrozek K, Auer H, Tanner SM, Guimond M, et al. Interestingly, this resulted in a significant increase in NF-B p65 binding to R4 of the ICAM-1 promoter (Fig. Loss of endothelial CFTR drives barrier failure and edema formation in lung infection and can be targeted by CFTR potentiation. N.D. designed and performed invivo experiments, analyzed, and interpreted results. See supplemental Table S1 for oligonucleotide sequences. In EC, Erg expression is down-regulated by inflammatory stimuli (9, 13); this is in contrast with ETS factors including Ets-1, Ets-2, and ESE-1, whose expression is increased after treatment with agents such as IL-1, TNF-, angiotensin II, or thrombin (9, 17, 5459). Only siRNA to Erg caused a significant increase in ICAM-1 mRNA levels (Fig. EMSA carried out with the EBS 181 oligonucleotide showed the formation of three specific complexes with nuclear proteins from resting HUVEC (Fig. (A) Representative whole mount images of E10.5, (B) Endomucin staining of blood vessels in E10.5, (C) Isolectin B4 staining of postnatal day 6 retinas from. Epub 2017 Jan 13. Mice were administered Tamoxifen (50g per mouse; Sigma) by intraperitoneal injection (IP) at postnatal (P) day 1, P2 and P3. Genome-wide analysis of the zebrafish ETS family identifies three genes required for hemangioblast differentiation or angiogenesis. The following day, cells were transduced with 100 multiplicity of infection of IB Super Repressor Adenovirus (AdIBSR) (14) or AdLacZ in serum-free M199 medium for 2 h before replacing with complete M199 medium. A similar pattern was observed with a different anti-Erg antibody (data not shown), suggesting that the absence of a supershift is not due to a technical issue related to a single antibody but possibly because the new complex is too large to be identified by this method. All graphical data are SEM, Pericyte recruitment is a critical step in vascular stability and maturation, and lack of pericytes has been shown to cause increased permeability (. In addition, immunostains for CD31, CD34, and -smooth muscle actin (-SMA) were performed on all samples. official website and that any information you provide is encrypted HHS Vulnerability Disclosure, Help Dark-adapted ERGs recordings showing ERG responses to dim and bright flashes in Gnat1 / . EMSA reactions were run on a 0.5 Tris borate-EDTA (TBE) polyacrylamide gel and transferred to Hybond N+ nylon membrane (GE Healthcare, Amersham Biosciences) before detection. Cell 32, 8296 (2015). Don't show this again. The conditional inactivation of the beta-catenin gene in endothelial cells causes a defective vascular pattern and increased vascular fragility. Would you like email updates of new search results? HES6. designed and carried out invitro experiments, analyzed, interpreted, and conceptualized results, and wrote the manuscript. Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice. Nuclear extracts from resting HUVEC were incubated with biotinylated double-stranded oligonucleotides containing either the EBS 118 or 181 sequence. Deletion of ERG disrupts regulation of coagulation, leading to a. contributed to scientific discussion; H.G. and JavaScript. To specifically enrich our samples in endothelial cells and microglia, with their important role in dementia and white matter disease, 22 we immunolabeled the nuclei using an antibody against ERG . Results show additional HLA-DPB1 polymorphism in exons 1, 3, 4 and 5 and the 5' and 3'-UTR. 3B, lanes 46, arrows), suggesting that the anti-NF-B p65 antibody binds to the protein-oligonucleotide complex at this site. The second study investigated the response of microvascular and macrovascular endothelial cells to TNF-, interferon , or IL-4 (18). In the presence of bFGF (, Matrigel containing bFGF or VEGF with adenovirus expressing either Lacz (Ad.Lacz) or ERG (Ad.ERG) was injected into C57BL6 mice. This article demonstrates the importance of FLI1 in endothelial cell function, and suggests a link with the development of vascular maladies. We carried out an initial screen to identify any overlap between genes up-regulated following Erg inhibition and published datasets held in the Broad Institute curated GSEA MSigDB database. Transcription factors are involved in a large number of human diseases such as cancers for which they account for about 20% of all oncogenes identified so far. The endothelium forms an interface between circulating blood or lymph in the lumen and the rest of the vessel wall. Proc Natl Acad Sci U S A. Cardiac endothelium communicates closely with adjacent cardiac cells by multiple cytokines and plays critical roles in regulating fibroblasts proliferation, activation, and collagen synthesis during cardiac fibrosis. In this study we showed that Ets-2, Fli-1, and GABP, although constitutively expressed in EC, do not repress ICAM-1 expression. First, we showed that Erg inhibition in HUVEC significantly increased the mRNA levels of cIAP2 (Fig. The Author(s), under exclusive licence to Springer Nature B.V. part of Springer Nature. At the junctions, VE-cadherin binds -catenin, protecting it from degradation; this interaction is required for the control of junction stabilization (. I.M.A. (C) -catenin (-cat; green) and VE-cadherin (VEC; red) staining of FITC-conjugated siCtrl and siERG (FITC; purple) treated HUVEC (n= 3). Progressive loss of retinal ganglion cell axons - manifests as irreversible . Vascular lesions (hemangioma and lymphangioma) are very rare in the testis, their diagnosis can be easily ruled out by the negative immunostaining with mesothelial markers and their positive staining with endothelial markers like CD34 (cluster of differentiation 34), CD31, ERG, factor VIII-related antigen [13, 10, 11]. Erg is the most abundantly expressed ETS factor in resting ECs and its expression, although not uniquely endothelial, is restricted mostly to the hematopoietic/endothelial lineage. The site is secure. Endothelial cells are simple squamous cells which line the entire vascular system (including lymphatic channels). Nat Rev Cancer. Acta Neuropathol. These markers and other endothelial markers are also discussed in Chapter 13. Would you like email updates of new search results? In addition, we also used GSEA to compare the Erg-regulated genes with the data from a recent ChIP seq analysis, which identified regulatory sequences bound by NF-B p65 in LPS-treated macrophages (19). Abstract:Endothelial cells (ECs) across ages and tissues are highly heterogeneous in developmental origins, structures, functions, and cellular plasticity. Data are shown as fold change over IgG (n= 3). These parent stem cells, ESCs, give rise to progenitor cells, which are . Keywords: 5D, panel i), confirming that cIAP2 is repressed by Erg in resting HUVEC. Blood. For Apelin expression assay, cells were cultured in the presence of both 4-hydroxytamoxifen and recombinant human VEGF. Blockade of ERG using siRNA completely interferes with EC lumen formation. The role of Erg as a repressor of inflammation is in contrast to that of other ETS factors, which have previously been shown to act synergistically with NF-B in promoting inflammatory gene expression. The following day, siRNA (10 nm) was mixed with AtuFect01 lipid (1 g/ml, Silence Therapeutics) at 5 times concentration in Opti-MEM (Invitrogen), then added to cells for 24 or 48 h. Erg overexpression was carried out using a V5-tagged Erg-3 adenovirus (AdErg), as described previously (12). A., Taborsky G. J., Jr., Chansky H. A. 2014. The clinical relevance of Erg in repressing EC activation is supported by its pattern of expression in atherosclerotic plaques: Erg is expressed in the healthy endothelium of human coronary artery but is absent from the activated endothelium over inflammatory infiltrate in the plaque shoulder (12). For long time, with the exception of ligand-inducible nuclear receptors, transcription factors were considered as "undruggable" targets. Evidence for an inducible autoregulatory pathway, Obesity, inflammation, and atherosclerosis, Karim F. D., Urness L. D., Thummel C. S., Klemsz M. J., McKercher S. R., Celada A., Van Beveren C., Maki R. A., Gunther C. V., Nye J. (E) (Top) Representative whole mount images of E10.5. Transcription cofactor HES-6 is a protein that in humans is encoded by the HES6 gene. We show that constitutive deletion of endothelial ERG in the mouse embryo causes embryonic lethality with severe vascular disruption. (2006), Endoglin expression in the endothelium is regulated by Fli-1, Erg, and Elf-1 acting on the promoter and a 8-kb enhancer, McLaughlin F., Ludbrook V. J., Kola I., Campbell C. J., Randi A. M. (1999), Characterization of the tumor necrosis factor (TNF)- response elements in the human ICAM-2 promoter, Birdsey G. M., Dryden N. H., Amsellem V., Gebhardt F., Sahnan K., Haskard D. O., Dejana E., Mason J. C., Randi A. M. (2008), Transcription factor Erg regulates angiogenesis and endothelial apoptosis through VE-cadherin, Deramaudt B. M., Remy P., Abraham N. G. (1999), Up-regulation of human heme oxygenase gene expression by Ets family proteins, Sperone A., Dryden N. H., Birdsey G. M., Madden L., Johns M., Evans P. C., Mason J. C., Haskard D. O., Boyle J. J., Paleolog E. M., Randi A. M. (2011), The transcription factor Erg inhibits vascular inflammation by repressing NF-B activation and proinflammatory gene expression in endothelial cells, Yuan L., Nikolova-Krstevski V., Zhan Y., Kondo M., Bhasin M., Varghese L., Yano K., Carman C. V., Aird W. C., Oettgen P. (2009), Anti-inflammatory effects of the ETS factor ERG in endothelial cells are mediated through transcriptional repression of the interleukin-8 gene, Brockman J. FOIA Accessibility Thank you for visiting nature.com. The ETS (E-26 transformation-specific) transcription factor ERG (ETS-related gene) is essential for endothelial homeostasis, driving expression of lineage genes and repressing proinflammatory genes. View chapter Purchase book Endothelium Alberto Mantovani, Elisabetta Dejana, in Encyclopedia of Immunology (Second Edition), 1998 PECAM-1 Endothelial stem cells (ESCs) are one of three types of stem cells found in bone marrow.They are multipotent, which describes the ability to give rise to many cell types, whereas a pluripotent stem cell can give rise to all types. AC, GSEA was carried out using standard settings. 1D). data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAKAAAAB4CAYAAAB1ovlvAAAAAXNSR0IArs4c6QAAAnpJREFUeF7t17Fpw1AARdFv7WJN4EVcawrPJZeeR3u4kiGQkCYJaXxBHLUSPHT/AaHTvu . 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