Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells

A H Sikkema; E S J M de Bont; G Molema; A Dimberg; P J Zwiers; S H Diks; E W Hoving; W A Kamps; M P Peppelenbosch; W F A den Dunnen

doi:10.1111/j.1365-2990.2011.01160.x

Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells

A H Sikkema, E S J M de Bont, G Molema, A Dimberg, P J Zwiers, S H Diks, E W Hoving, W A Kamps, M P Peppelenbosch, W F A den Dunnen

Onderzoeksoutput: Bijdrage aan tijdschrift › Artikel › peer review

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AIMS: Tumours depend on angiogenesis for enhanced tumour cell survival and progression. Vascular endothelial growth factor receptor (VEGFR) signalling plays a major part in this process. Previously, we evaluated tyrosine kinase activity in paediatric brain tumour tissue lysates using a peptide microarray containing 144 different tyrosine kinase peptide substrates. When applied to paediatric pilocytic astrocytoma tissue, this analysis revealed extensive phosphorylation of VEGFR-derived peptides. The aim of the current study was to validate this result and determine the presence of VEGFR-2 activity in paediatric pilocytic astrocytoma as the main VEGFR in terms of mitogenic signalling. In addition, the localization of VEGFR1-3 mRNA expression was assessed.

METHODS: VEGFR-2 phosphorylation was determined by adopting a proximity ligation assay approach. Enrichment of endothelial markers and VEGFRs in tumour endothelium was determined by quantitative polymerase chain reaction (qPCR) analysis of laser-microdissected blood vessels.

RESULTS: Proximity ligation assays on tumour cryosections showed the presence of phosphorylation of VEGFR-2, which primarily localized to vascular endothelium. qPCR analysis of endothelial markers and VEGFRs showed a 13.6-fold average enrichment of VEGFR-2 expression in the laser-microdissected endothelium compared to whole tumour. Also the expression of VEGFR-1 and -3 was highly enriched in the endothelium fraction with an average fold-enrichment of 16.5 and 50.8 respectively.

CONCLUSIONS: Phosphorylated VEGFR-2 is detected on endothelial cells in paediatric pilocytic astrocytoma. Furthermore, endothelial cells are the main source of VEGFR1-3 mRNA expression. This suggests a crucial role for VEGF/VEGFR-induced angiogenesis in the progression and maintenance of these tumours.

Originele taal-2	Engels
Pagina's (van-tot)	538-48
Aantal pagina's	11
Tijdschrift	Neuropathology and Applied Neurobiology
Volume	37
Nummer van het tijdschrift	5
DOI's	https://doi.org/10.1111/j.1365-2990.2011.01160.x
Status	Gepubliceerd - aug. 2011
Extern gepubliceerd	Ja

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10.1111/j.1365-2990.2011.01160.x

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Sikkema, A. H., de Bont, E. S. J. M., Molema, G., Dimberg, A., Zwiers, P. J., Diks, S. H., Hoving, E. W., Kamps, W. A., Peppelenbosch, M. P., & den Dunnen, W. F. A. (2011). Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells. Neuropathology and Applied Neurobiology, 37(5), 538-48. https://doi.org/10.1111/j.1365-2990.2011.01160.x

@article{adf95c26a7294dfc97b64cd7240a98d2,

title = "Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells",

abstract = "AIMS: Tumours depend on angiogenesis for enhanced tumour cell survival and progression. Vascular endothelial growth factor receptor (VEGFR) signalling plays a major part in this process. Previously, we evaluated tyrosine kinase activity in paediatric brain tumour tissue lysates using a peptide microarray containing 144 different tyrosine kinase peptide substrates. When applied to paediatric pilocytic astrocytoma tissue, this analysis revealed extensive phosphorylation of VEGFR-derived peptides. The aim of the current study was to validate this result and determine the presence of VEGFR-2 activity in paediatric pilocytic astrocytoma as the main VEGFR in terms of mitogenic signalling. In addition, the localization of VEGFR1-3 mRNA expression was assessed.METHODS: VEGFR-2 phosphorylation was determined by adopting a proximity ligation assay approach. Enrichment of endothelial markers and VEGFRs in tumour endothelium was determined by quantitative polymerase chain reaction (qPCR) analysis of laser-microdissected blood vessels.RESULTS: Proximity ligation assays on tumour cryosections showed the presence of phosphorylation of VEGFR-2, which primarily localized to vascular endothelium. qPCR analysis of endothelial markers and VEGFRs showed a 13.6-fold average enrichment of VEGFR-2 expression in the laser-microdissected endothelium compared to whole tumour. Also the expression of VEGFR-1 and -3 was highly enriched in the endothelium fraction with an average fold-enrichment of 16.5 and 50.8 respectively.CONCLUSIONS: Phosphorylated VEGFR-2 is detected on endothelial cells in paediatric pilocytic astrocytoma. Furthermore, endothelial cells are the main source of VEGFR1-3 mRNA expression. This suggests a crucial role for VEGF/VEGFR-induced angiogenesis in the progression and maintenance of these tumours.",

keywords = "Astrocytoma/metabolism, Child, Endothelial Cells/metabolism, Fluorescent Antibody Technique, Humans, Lasers, Microdissection, Neovascularization, Pathologic/metabolism, Phosphorylation, Protein Array Analysis, RNA, Messenger/analysis, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction/physiology, Vascular Endothelial Growth Factor Receptor-1/analysis, Vascular Endothelial Growth Factor Receptor-2/analysis, Vascular Endothelial Growth Factor Receptor-3/analysis",

author = "Sikkema, {A H} and {de Bont}, {E S J M} and G Molema and A Dimberg and Zwiers, {P J} and Diks, {S H} and Hoving, {E W} and Kamps, {W A} and Peppelenbosch, {M P} and {den Dunnen}, {W F A}",

note = "{\textcopyright} 2011 The Authors. Neuropathology and Applied Neurobiology {\textcopyright} 2011 British Neuropathological Society.",

year = "2011",

month = aug,

doi = "10.1111/j.1365-2990.2011.01160.x",

language = "English",

volume = "37",

pages = "538--48",

journal = " Neuropathology and Applied Neurobiology",

issn = "0305-1846",

publisher = "Wiley-Blackwell Publishing Ltd",

number = "5",

}

Sikkema, AH, de Bont, ESJM, Molema, G, Dimberg, A, Zwiers, PJ, Diks, SH, Hoving, EW, Kamps, WA, Peppelenbosch, MP & den Dunnen, WFA 2011, 'Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells', Neuropathology and Applied Neurobiology, vol. 37, nr. 5, blz. 538-48. https://doi.org/10.1111/j.1365-2990.2011.01160.x

TY - JOUR

T1 - Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells

AU - Sikkema, A H

AU - de Bont, E S J M

AU - Molema, G

AU - Dimberg, A

AU - Zwiers, P J

AU - Diks, S H

AU - Hoving, E W

AU - Kamps, W A

AU - Peppelenbosch, M P

AU - den Dunnen, W F A

PY - 2011/8

Y1 - 2011/8

N2 - AIMS: Tumours depend on angiogenesis for enhanced tumour cell survival and progression. Vascular endothelial growth factor receptor (VEGFR) signalling plays a major part in this process. Previously, we evaluated tyrosine kinase activity in paediatric brain tumour tissue lysates using a peptide microarray containing 144 different tyrosine kinase peptide substrates. When applied to paediatric pilocytic astrocytoma tissue, this analysis revealed extensive phosphorylation of VEGFR-derived peptides. The aim of the current study was to validate this result and determine the presence of VEGFR-2 activity in paediatric pilocytic astrocytoma as the main VEGFR in terms of mitogenic signalling. In addition, the localization of VEGFR1-3 mRNA expression was assessed.METHODS: VEGFR-2 phosphorylation was determined by adopting a proximity ligation assay approach. Enrichment of endothelial markers and VEGFRs in tumour endothelium was determined by quantitative polymerase chain reaction (qPCR) analysis of laser-microdissected blood vessels.RESULTS: Proximity ligation assays on tumour cryosections showed the presence of phosphorylation of VEGFR-2, which primarily localized to vascular endothelium. qPCR analysis of endothelial markers and VEGFRs showed a 13.6-fold average enrichment of VEGFR-2 expression in the laser-microdissected endothelium compared to whole tumour. Also the expression of VEGFR-1 and -3 was highly enriched in the endothelium fraction with an average fold-enrichment of 16.5 and 50.8 respectively.CONCLUSIONS: Phosphorylated VEGFR-2 is detected on endothelial cells in paediatric pilocytic astrocytoma. Furthermore, endothelial cells are the main source of VEGFR1-3 mRNA expression. This suggests a crucial role for VEGF/VEGFR-induced angiogenesis in the progression and maintenance of these tumours.

AB - AIMS: Tumours depend on angiogenesis for enhanced tumour cell survival and progression. Vascular endothelial growth factor receptor (VEGFR) signalling plays a major part in this process. Previously, we evaluated tyrosine kinase activity in paediatric brain tumour tissue lysates using a peptide microarray containing 144 different tyrosine kinase peptide substrates. When applied to paediatric pilocytic astrocytoma tissue, this analysis revealed extensive phosphorylation of VEGFR-derived peptides. The aim of the current study was to validate this result and determine the presence of VEGFR-2 activity in paediatric pilocytic astrocytoma as the main VEGFR in terms of mitogenic signalling. In addition, the localization of VEGFR1-3 mRNA expression was assessed.METHODS: VEGFR-2 phosphorylation was determined by adopting a proximity ligation assay approach. Enrichment of endothelial markers and VEGFRs in tumour endothelium was determined by quantitative polymerase chain reaction (qPCR) analysis of laser-microdissected blood vessels.RESULTS: Proximity ligation assays on tumour cryosections showed the presence of phosphorylation of VEGFR-2, which primarily localized to vascular endothelium. qPCR analysis of endothelial markers and VEGFRs showed a 13.6-fold average enrichment of VEGFR-2 expression in the laser-microdissected endothelium compared to whole tumour. Also the expression of VEGFR-1 and -3 was highly enriched in the endothelium fraction with an average fold-enrichment of 16.5 and 50.8 respectively.CONCLUSIONS: Phosphorylated VEGFR-2 is detected on endothelial cells in paediatric pilocytic astrocytoma. Furthermore, endothelial cells are the main source of VEGFR1-3 mRNA expression. This suggests a crucial role for VEGF/VEGFR-induced angiogenesis in the progression and maintenance of these tumours.

KW - Astrocytoma/metabolism

KW - Child

KW - Endothelial Cells/metabolism

KW - Fluorescent Antibody Technique

KW - Humans

KW - Lasers

KW - Microdissection

KW - Neovascularization, Pathologic/metabolism

KW - Phosphorylation

KW - Protein Array Analysis

KW - RNA, Messenger/analysis

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Signal Transduction/physiology

KW - Vascular Endothelial Growth Factor Receptor-1/analysis

KW - Vascular Endothelial Growth Factor Receptor-2/analysis

KW - Vascular Endothelial Growth Factor Receptor-3/analysis

U2 - 10.1111/j.1365-2990.2011.01160.x

DO - 10.1111/j.1365-2990.2011.01160.x

M3 - Article

C2 - 21208252

SN - 0305-1846

VL - 37

SP - 538

EP - 548

JO - Neuropathology and Applied Neurobiology

JF - Neuropathology and Applied Neurobiology

IS - 5

ER -

Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells

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