Skip to main content
SpringerLink
Log in

Ets-1 transcription is required in tissue factor driven microvessel formation and stabilization

  • Original Paper
  • Published:
Angiogenesis Aims and scope Submit manuscript

Abstract

Tissue factor (TF) has well-recognized roles as initiator of blood coagulation as well as an intracellular signaling receptor. TF signaling regulates gene transcription and protein translation. Recently, we have shown that TF-induced mature neovessel formation is ultimately driven by CCL2 expression. However, the signaling process induced by TF to promote microvessel formation remains to be determined. This study was designed with the objective to investigate the mechanisms involved in TF-induced neovessel formation. Here, we have identified that Ets-1 expression is a downstream effector of TF signaling. TF-siRNA induced a highly significant reduction in Ets-1 expression levels and in Ets-1/DNA binding while inducing abrogation of microvessel formation. Activation of Ets-1 rescued the effect of TF inhibition and restored microvessel formation confirming the critical role of Ets-1 in TF-induced angiogenesis. VE-cadherin expression, a key regulator of endothelial intercellular junctions, and an Ets-1 target molecule was dependent of TF-inhibition. We show that TF signals through ERK1/2 to activate Ets-1 and induce CCL2 gene expression by binding to its promoter region. We conclude that endothelial cell TF signals through ERK1/2 and Ets-1 to trigger microvessel formation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Davie EW, Fujikawa K, Kisiel W (1991) The coagulation cascade: initiation, maintenance, and regulation. Biochemistry 30:10363–10370

    Article  PubMed  CAS  Google Scholar 

  2. Edgington TS, Mackman N, Brand K, Ruf W (1991) The structural biology of expression and function of tissue factor. Thromb Haemost 66:67–79

    PubMed  CAS  Google Scholar 

  3. Tremoli E, Camera M, Toschi V, Colli S (1999) Tissue factor in atherosclerosis. Atherosclerosis 144:273–283

    Article  PubMed  CAS  Google Scholar 

  4. Lopez-Pedrera C, Barbarroja N, Dorado G, Siendones E, Velasco F (2006) Tissue factor as an effector of angiogenesis and tumor progression in hematological malignancies. Leukemia 20:1331–1340. doi:10.1038/sj.leu.2404264

    Article  PubMed  CAS  Google Scholar 

  5. Juan-Babot JO, Martinez-Gonzalez J, Berrozpe M, Badimon L (2003) Neovascularization in human coronary arteries with lesions of different severity. Rev Esp Cardiol 56:978–986

    Article  PubMed  Google Scholar 

  6. Moreno PR, Purushothaman KR, Fuster V, Echeverri D, Truszczynska H, Sharma SK, Badimon JJ, O’Connor WN (2004) Plaque neovascularization is increased in ruptured atherosclerotic lesions of human aorta: implications for plaque vulnerability. Circulation 110:2032–2038. doi:10.1161/01.CIR.0000143233.87854.23

    Article  PubMed  Google Scholar 

  7. Virmani R, Kolodgie FD, Burke AP, Finn AV, Gold HK, Tulenko TN, Wrenn SP, Narula J (2005) Atherosclerotic plaque progression and vulnerability to rupture: angiogenesis as a source of intraplaque hemorrhage. Arterioscler Thromb Vasc Biol 25:2054–2061. doi:10.1161/01.ATV.0000178991.71605.18

    Article  PubMed  CAS  Google Scholar 

  8. Carmeliet P (2003) Angiogenesis in health and disease. Nat Med 9:653–660. doi:10.1038/nm0603-653

    Article  PubMed  CAS  Google Scholar 

  9. Arderiu G, Pena E, Aledo R, Juan-Babot O, Badimon L (2011) Tissue factor regulates microvessel formation and stabilization by induction of chemokine (C–C motif) ligand 2 expression. Arterioscler Thromb Vasc Biol 31:2607–2615. doi:10.1161/ATVBAHA.111.233536

    Article  PubMed  CAS  Google Scholar 

  10. Wasylyk B, Hahn SL, Giovane A (1993) The Ets family of transcription factors. Eur J Biochem 211:7–18

    Article  PubMed  CAS  Google Scholar 

  11. Wernert N, Raes MB, Lassalle P, Dehouck MP, Gosselin B, Vandenbunder B, Stehelin D (1992) c-Ets1 proto-oncogene is a transcription factor expressed in endothelial cells during tumor vascularization and other forms of angiogenesis in humans. Am J Pathol 140:119–127

    PubMed  CAS  Google Scholar 

  12. Zhan Y, Brown C, Maynard E, Anshelevich A, Ni W, Ho IC, Oettgen P (2005) Ets-1 is a critical regulator of Ang II-mediated vascular inflammation and remodeling. J Clin Invest 115:2508–2516. doi:10.1172/JCI24403

    Article  PubMed  CAS  Google Scholar 

  13. Aoki T, Kataoka H, Nishimura M, Ishibashi R, Morishita R, Miyamoto S (2010) Ets-1 promotes the progression of cerebral aneurysm by inducing the expression of MCP-1 in vascular smooth muscle cells. Gene Ther 17:1117–1123. doi:10.1038/gt.2010.60

    Article  PubMed  CAS  Google Scholar 

  14. Ades EW, Candal FJ, Swerlick RA, George VG, Summers S, Bosse DC, Lawley TJ (1992) HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J Invest Dermatol 99:683–690

    Article  PubMed  CAS  Google Scholar 

  15. Camino-Lopez S, Badimon L, Gonzalez A, Canals D, Pena E, Llorente-Cortes V (2009) Aggregated low density lipoprotein induces tissue factor by inhibiting sphingomyelinase activity in human vascular smooth muscle cells. J Thromb Haemost 7:2137–2146. doi:10.1111/j.1538-7836.2009.03638.x

    Article  PubMed  CAS  Google Scholar 

  16. Arderiu G, Cuevas I, Chen A, Carrio M, East L, Boudreau NJ (2007) HoxA5 stabilizes adherens junctions via increased Akt1. Cell Adh Migr 1:185–195

    Article  PubMed  Google Scholar 

  17. Kitahara T, Hiromura K, Ikeuchi H, Yamashita S, Kobayashi S, Kuroiwa T, Kaneko Y, Ueki K, Nojima Y (2005) Mesangial cells stimulate differentiation of endothelial cells to form capillary-like networks in a three-dimensional culture system. Nephrol Dial Transpl 20:42–49. doi:10.1093/ndt/gfh572

    Article  CAS  Google Scholar 

  18. Otero-Vinas M, Llorente-Cortes V, Pena E, Padro T, Badimon L (2007) Aggregated low density lipoproteins decrease metalloproteinase-9 expression and activity in human coronary smooth muscle cells. Atherosclerosis 194:326–333

    Article  PubMed  CAS  Google Scholar 

  19. Choi YB, Nicholas J (2010) Induction of angiogenic chemokine CCL2 by human herpesvirus 8 chemokine receptor. Virology 397:369–378. doi:10.1016/j.virol.2009.11.024

    Article  PubMed  CAS  Google Scholar 

  20. Adini A, Fainaru O, Udagawa T, Connor KM, Folkman J, D’Amato RJ (2009) Matrigel cytometry: a novel method for quantifying angiogenesis in vivo. J Immunol Methods 342:78–81. doi:10.1016/j.jim.2008.11.016

    Article  PubMed  CAS  Google Scholar 

  21. Carrio M, Arderiu G, Myers C, Boudreau NJ (2005) Homeobox D10 induces phenotypic reversion of breast tumor cells in a three-dimensional culture model. Cancer Res 65:7177–7185. doi:10.1158/0008-5472.CAN-04-1717

    Article  PubMed  CAS  Google Scholar 

  22. Oikawa T, Yamada T (2003) Molecular biology of the Ets family of transcription factors. Gene 303:11–34

    Article  PubMed  CAS  Google Scholar 

  23. Pradier A, Ettelaie C (2008) The influence of exogenous tissue factor on the regulators of proliferation and apoptosis in endothelial cells. J Vasc Res 45:19–32. doi:10.1159/000109074

    Article  PubMed  CAS  Google Scholar 

  24. Gory S, Dalmon J, Prandini MH, Kortulewski T, de Launoit Y, Huber P (1998) Requirement of a GT box (Sp1 site) and two Ets binding sites for vascular endothelial cadherin gene transcription. J Biol Chem 273:6750–6755

    Article  PubMed  CAS  Google Scholar 

  25. Lelievre E, Mattot V, Huber P, Vandenbunder B, Soncin F (2000) ETS1 lowers capillary endothelial cell density at confluence and induces the expression of VE-cadherin. Oncogene 19:2438–2446. doi:10.1038/sj.onc.1203563

    Article  PubMed  CAS  Google Scholar 

  26. Vittet D, Buchou T, Schweitzer A, Dejana E, Huber P (1997) Targeted null-mutation in the vascular endothelial-cadherin gene impairs the organization of vascular-like structures in embryoid bodies. Proc Natl Acad Sci USA 94:6273–6278

    Article  PubMed  CAS  Google Scholar 

  27. Groupp ER, Donovan-Peluso M (1996) Lipopolysaccharide induction of THP-1 cells activates binding of c-Jun, Ets, and Egr-1 to the tissue factor promoter. J Biol Chem 271:12423–12430

    Article  PubMed  CAS  Google Scholar 

  28. Sato Y (2001) Role of ETS family transcription factors in vascular development and angiogenesis. Cell Struct Funct 26:19–24

    Article  PubMed  CAS  Google Scholar 

  29. Dejana E, Taddei A, Randi AM (2007) Foxs and Ets in the transcriptional regulation of endothelial cell differentiation and angiogenesis. Biochim Biophys Acta 1775:298–312. doi:10.1016/j.bbcan.2007.05.003

    PubMed  CAS  Google Scholar 

  30. Vandenbunder B, Pardanaud L, Jaffredo T, Mirabel MA, Stehelin D (1989) Complementary patterns of expression of c-ets 1, c-myb and c-myc in the blood-forming system of the chick embryo. Development 107:265–274

    PubMed  CAS  Google Scholar 

  31. Sato Y, Abe M, Tanaka K, Iwasaka C, Oda N, Kanno S, Oikawa M, Nakano T, Igarashi T (2000) Signal transduction and transcriptional regulation of angiogenesis. Adv Exp Med Biol 476:109–115

    Article  PubMed  CAS  Google Scholar 

  32. Iwasaka C, Tanaka K, Abe M, Sato Y (1996) Ets-1 regulates angiogenesis by inducing the expression of urokinase-type plasminogen activator and matrix metalloproteinase-1 and the migration of vascular endothelial cells. J Cell Physiol 169:522–531. doi:10.1002/(SICI)1097-4652(199612)169:3<522:AID-JCP12>3.0.CO;2-7

    Article  PubMed  CAS  Google Scholar 

  33. Tanaka K, Abe M, Sato Y (1999) Roles of extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase in the signal transduction of basic fibroblast growth factor in endothelial cells during angiogenesis. Jpn J Cancer Res 90:647–654

    Article  PubMed  CAS  Google Scholar 

  34. Oda N, Abe M, Sato Y (1999) ETS-1 converts endothelial cells to the angiogenic phenotype by inducing the expression of matrix metalloproteinases and integrin beta3. J Cell Physiol 178:121–132. doi:10.1002/(SICI)1097-4652(199902)178:2<121:AID-JCP1>3.0.CO;2-F

    Article  PubMed  CAS  Google Scholar 

  35. Corada M, Mariotti M, Thurston G, Smith K, Kunkel R, Brockhaus M, Lampugnani MG, Martin-Padura I, Stoppacciaro A, Ruco L, McDonald DM, Ward PA, Dejana E (1999) Vascular endothelial-cadherin is an important determinant of microvascular integrity in vivo. Proc Natl Acad Sci USA 96:9815–9820

    Article  PubMed  CAS  Google Scholar 

  36. Falcon BL, Hashizume H, Koumoutsakos P, Chou J, Bready JV, Coxon A, Oliner JD, McDonald DM (2009) Contrasting actions of selective inhibitors of angiopoietin-1 and angiopoietin-2 on the normalization of tumor blood vessels. Am J Pathol 175:2159–2170. doi:10.2353/ajpath.2009.090391

    Article  PubMed  CAS  Google Scholar 

  37. Russell L, Garrett-Sinha LA (2010) Transcription factor Ets-1 in cytokine and chemokine gene regulation. Cytokine 51:217–226. doi:10.1016/j.cyto.2010.03.006

    Article  PubMed  CAS  Google Scholar 

  38. Yang BS, Hauser CA, Henkel G, Colman MS, Van Beveren C, Stacey KJ, Hume DA, Maki RA, Ostrowski MC (1996) Ras-mediated phosphorylation of a conserved threonine residue enhances the transactivation activities of c-Ets1 and c-Ets2. Mol Cell Biol 16:538–547

    PubMed  CAS  Google Scholar 

  39. Chen YF, Shin SJ, Lin SR (2005) Ets1 was significantly activated by ERK1/2 in mutant K-ras stably transfected human adrenocortical cells. DNA Cell Biol 24:126–132. doi:10.1089/dna.2005.24.126

    Article  PubMed  CAS  Google Scholar 

  40. Goetze S, Kintscher U, Kaneshiro K, Meehan WP, Collins A, Fleck E, Hsueh WA, Law RE (2001) TNFalpha induces expression of transcription factors c-fos, Egr-1, and Ets-1 in vascular lesions through extracellular signal-regulated kinases 1/2. Atherosclerosis 159:93–101

    Article  PubMed  CAS  Google Scholar 

  41. Poulsen LK, Jacobsen N, Sorensen BB, Bergenhem NC, Kelly JD, Foster DC, Thastrup O, Ezban M, Petersen LC (1998) Signal transduction via the mitogen-activated protein kinase pathway induced by binding of coagulation factor VIIa to tissue factor. J Biol Chem 273:6228–6232

    Article  PubMed  CAS  Google Scholar 

  42. Arderiu G, Pena E, Aledo R, Badimon L (2012) Tissue factor-akt signaling triggers microvessel formation. J Thromb Haemost. doi:10.1111/j.1538-7836.2012.04848.x

    Google Scholar 

Download references

Acknowledgments

We thank O. Garcia and M. Pescador for their excellent technical assistance. This work was supported in part by grants from Ministry of Science and Education of Spain (SAF2010/16549), and Instituto de Salud Carlos III (CIBEROBN-CB06/03) (to LB), and (CP07/00224) (to GA). We thank the Fundación de Investigación Cardiovascular and the Fundación Jesus Serra for their support.

Conflict of interest

The authors declared no conflict of interest.

Author information

Authors and Affiliations

  1. Cardiovascular Research Center (CSIC-ICCC), Hospital de Sant Pau (UAB), IIB-Sant Pau. CiberOBN, Instituto de Salut Carlos III, C/ Sant Antoni Mª Claret 167, 08025, Barcelona, Spain

    Gemma Arderiu, Esther Peña, Rosa Aledo, Sonia Espinosa & Lina Badimon

Authors
  1. Gemma Arderiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Esther Peña
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rosa Aledo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sonia Espinosa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lina Badimon
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lina Badimon.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 4449 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arderiu, G., Peña, E., Aledo, R. et al. Ets-1 transcription is required in tissue factor driven microvessel formation and stabilization. Angiogenesis 15, 657–669 (2012). https://doi.org/10.1007/s10456-012-9293-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10456-012-9293-x

Keywords

Search

Navigation