Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitvo

Gianluca Baldanzi, Stefania Mitola, Santina Cutrupi, Nicoletta Filigheddu, Wim J. Van Blitterswijk, Fabiola Sinigaglia, Federico Bussolino, Andrea Graziani

Research output: Contribution to journal › Article

Abstract

Vascular endothelial growth factor-A (VEGF-A) promotes angiogenesis by stimulating migration, proliferation and organization of endothelium, through the activation of signaling pathways involving Src tyrosine kinase. As we had previously shown that Src-mediated activation of diacylglycerol kinase-α (Dgk-α) is required for hepatocytes growth factor-stimulated cell migration, we asked whether Dgk-α is involved in the transduction of angiogenic signaling. In PAE-KDR cells, an endothelial-derived cell line expressing VEGFR-2, VEGF-A165, stimulates the enzymatic activity of Dgk-α: activation is inhibited by R59949, an isoform-specific Dgk inhibitor, and is dependent on Src tyrosine kinase, with which Dgk-α forms a complex. Conversely in HUVEC, VEGF-A165-induced activation of Dgk is only partially sensitive to R59949, suggesting that also other isoforms may be activated, albeit still dependent on Src tyrosine kinase. Specific inhibition of Dgk-α, obtained in both cells by R59949 and in PAE-KDR by expression of Dgk-α dominant-negative mutant, impairs VEGF-A165-dependent chemotaxis, proliferation and in vitro angiogenesis. In addition, in HUVEC, specific downregulation of Dgk-α by siRNA impairs in vitro angiogenesis on matrigel, further suggesting the requirement for Dgk-α in angiogenic signaling in HUVEC. Thus, we propose that activation of Dgk-α generates a signal essential for both proliferative and migratory response to VEGF-A 165, suggesting that it may constitute a novel pharmacological target for angiogenesis control.

Original languageEnglish
Pages (from-to)4828-4838
Number of pages11
JournalOncogene
Volume23
Issue number28
DOIs
Publication statusPublished - Jun 17 2004

Fingerprint

Diacylglycerol Kinase
Vascular Endothelial Growth Factor A
R 59949
src-Family Kinases
Protein Isoforms
Vascular Endothelial Growth Factor Receptor-2
Hepatocyte Growth Factor
Chemotaxis
Small Interfering RNA
Endothelium
Cell Movement
Down-Regulation
Endothelial Cells
Pharmacology
Cell Line

Keywords

  • Angiogenesis
  • Diacylglycerol kinase
  • Phosphatidic acid
  • Src
  • VEGF

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Baldanzi, G., Mitola, S., Cutrupi, S., Filigheddu, N., Van Blitterswijk, W. J., Sinigaglia, F., ... Graziani, A. (2004). Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitvo. Oncogene, 23(28), 4828-4838. https://doi.org/10.1038/sj.onc.1207633

Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitvo. / Baldanzi, Gianluca; Mitola, Stefania; Cutrupi, Santina; Filigheddu, Nicoletta; Van Blitterswijk, Wim J.; Sinigaglia, Fabiola; Bussolino, Federico; Graziani, Andrea.

In: Oncogene, Vol. 23, No. 28, 17.06.2004, p. 4828-4838.

Research output: Contribution to journal › Article

Baldanzi, G, Mitola, S, Cutrupi, S, Filigheddu, N, Van Blitterswijk, WJ, Sinigaglia, F, Bussolino, F & Graziani, A 2004, 'Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitvo', Oncogene, vol. 23, no. 28, pp. 4828-4838. https://doi.org/10.1038/sj.onc.1207633
Baldanzi G, Mitola S, Cutrupi S, Filigheddu N, Van Blitterswijk WJ, Sinigaglia F et al. Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitvo. Oncogene. 2004 Jun 17;23(28):4828-4838. https://doi.org/10.1038/sj.onc.1207633
Baldanzi, Gianluca ; Mitola, Stefania ; Cutrupi, Santina ; Filigheddu, Nicoletta ; Van Blitterswijk, Wim J. ; Sinigaglia, Fabiola ; Bussolino, Federico ; Graziani, Andrea. / Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitvo. In: Oncogene. 2004 ; Vol. 23, No. 28. pp. 4828-4838.
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abstract = "Vascular endothelial growth factor-A (VEGF-A) promotes angiogenesis by stimulating migration, proliferation and organization of endothelium, through the activation of signaling pathways involving Src tyrosine kinase. As we had previously shown that Src-mediated activation of diacylglycerol kinase-α (Dgk-α) is required for hepatocytes growth factor-stimulated cell migration, we asked whether Dgk-α is involved in the transduction of angiogenic signaling. In PAE-KDR cells, an endothelial-derived cell line expressing VEGFR-2, VEGF-A165, stimulates the enzymatic activity of Dgk-α: activation is inhibited by R59949, an isoform-specific Dgk inhibitor, and is dependent on Src tyrosine kinase, with which Dgk-α forms a complex. Conversely in HUVEC, VEGF-A165-induced activation of Dgk is only partially sensitive to R59949, suggesting that also other isoforms may be activated, albeit still dependent on Src tyrosine kinase. Specific inhibition of Dgk-α, obtained in both cells by R59949 and in PAE-KDR by expression of Dgk-α dominant-negative mutant, impairs VEGF-A165-dependent chemotaxis, proliferation and in vitro angiogenesis. In addition, in HUVEC, specific downregulation of Dgk-α by siRNA impairs in vitro angiogenesis on matrigel, further suggesting the requirement for Dgk-α in angiogenic signaling in HUVEC. Thus, we propose that activation of Dgk-α generates a signal essential for both proliferative and migratory response to VEGF-A 165, suggesting that it may constitute a novel pharmacological target for angiogenesis control.",
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AU - Van Blitterswijk, Wim J.

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