Expression and activity of the calcitonin receptor family in a sample of primary human high-grade gliomas

Anna Ostrovskaya, Caroline Hick, Dana S. Hutchinson, Brett W. Stringer, Peter J. Wookey, Denise Wootten, Patrick M. Sexton, Sebastian G. B. Furness

Research output: Contribution to journalArticleResearchpeer-review

1 Citation (Scopus)

Abstract

Background: Glioblastoma (GBM) is the most common and aggressive type of primary brain cancer. With median survival of less than 15 months, identification and validation of new GBM therapeutic targets is of critical importance. Results: In this study we tested expression and performed pharmacological characterization of the calcitonin receptor (CTR) as well as other members of the calcitonin family of receptors in high-grade glioma (HGG) cell lines derived from individual patient tumours, cultured in defined conditions. Previous immunohistochemical data demonstrated CTR expression in GBM biopsies and we were able to confirm CALCR (gene encoding CTR) expression. However, as assessed by cAMP accumulation assay, only one of the studied cell lines expressed functional CTR, while the other cell lines have functional CGRP (CLR/RAMP1) receptors. The only CTR-expressing cell line (SB2b) showed modest coupling to the cAMP pathway and no activation of other known CTR signaling pathways, including ERK 1/2 and p38 MAP kinases, and Ca 2+ mobilization, supportive of low cell surface receptor expression. Exome sequencing data failed to account for the discrepancy between functional data and expression on the cell lines that do not respond to calcitonin(s) with no deleterious non-synonymous polymorphisms detected, suggesting that other factors may be at play, such as alternative splicing or rapid constitutive receptor internalisation. Conclusions: This study shows that GPCR signaling can display significant variation depending on cellular system used, and effects seen in model recombinant cell lines or tumour cell lines are not always reproduced in a more physiologically relevant system and vice versa.

Original languageEnglish
Article number157
Number of pages12
JournalBMC Cancer
Volume19
Issue number1
DOIs
Publication statusPublished - 18 Feb 2019

Keywords

  • Calcitonin receptor
  • Glioblastoma
  • GPCR
  • Signaling

Cite this

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title = "Expression and activity of the calcitonin receptor family in a sample of primary human high-grade gliomas",
abstract = "Background: Glioblastoma (GBM) is the most common and aggressive type of primary brain cancer. With median survival of less than 15 months, identification and validation of new GBM therapeutic targets is of critical importance. Results: In this study we tested expression and performed pharmacological characterization of the calcitonin receptor (CTR) as well as other members of the calcitonin family of receptors in high-grade glioma (HGG) cell lines derived from individual patient tumours, cultured in defined conditions. Previous immunohistochemical data demonstrated CTR expression in GBM biopsies and we were able to confirm CALCR (gene encoding CTR) expression. However, as assessed by cAMP accumulation assay, only one of the studied cell lines expressed functional CTR, while the other cell lines have functional CGRP (CLR/RAMP1) receptors. The only CTR-expressing cell line (SB2b) showed modest coupling to the cAMP pathway and no activation of other known CTR signaling pathways, including ERK 1/2 and p38 MAP kinases, and Ca 2+ mobilization, supportive of low cell surface receptor expression. Exome sequencing data failed to account for the discrepancy between functional data and expression on the cell lines that do not respond to calcitonin(s) with no deleterious non-synonymous polymorphisms detected, suggesting that other factors may be at play, such as alternative splicing or rapid constitutive receptor internalisation. Conclusions: This study shows that GPCR signaling can display significant variation depending on cellular system used, and effects seen in model recombinant cell lines or tumour cell lines are not always reproduced in a more physiologically relevant system and vice versa.",
keywords = "Calcitonin receptor, Glioblastoma, GPCR, Signaling",
author = "Anna Ostrovskaya and Caroline Hick and Hutchinson, {Dana S.} and Stringer, {Brett W.} and Wookey, {Peter J.} and Denise Wootten and Sexton, {Patrick M.} and Furness, {Sebastian G. B.}",
year = "2019",
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Expression and activity of the calcitonin receptor family in a sample of primary human high-grade gliomas. / Ostrovskaya, Anna; Hick, Caroline; Hutchinson, Dana S.; Stringer, Brett W.; Wookey, Peter J.; Wootten, Denise; Sexton, Patrick M.; Furness, Sebastian G. B.

In: BMC Cancer, Vol. 19, No. 1, 157, 18.02.2019.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Expression and activity of the calcitonin receptor family in a sample of primary human high-grade gliomas

AU - Ostrovskaya, Anna

AU - Hick, Caroline

AU - Hutchinson, Dana S.

AU - Stringer, Brett W.

AU - Wookey, Peter J.

AU - Wootten, Denise

AU - Sexton, Patrick M.

AU - Furness, Sebastian G. B.

PY - 2019/2/18

Y1 - 2019/2/18

N2 - Background: Glioblastoma (GBM) is the most common and aggressive type of primary brain cancer. With median survival of less than 15 months, identification and validation of new GBM therapeutic targets is of critical importance. Results: In this study we tested expression and performed pharmacological characterization of the calcitonin receptor (CTR) as well as other members of the calcitonin family of receptors in high-grade glioma (HGG) cell lines derived from individual patient tumours, cultured in defined conditions. Previous immunohistochemical data demonstrated CTR expression in GBM biopsies and we were able to confirm CALCR (gene encoding CTR) expression. However, as assessed by cAMP accumulation assay, only one of the studied cell lines expressed functional CTR, while the other cell lines have functional CGRP (CLR/RAMP1) receptors. The only CTR-expressing cell line (SB2b) showed modest coupling to the cAMP pathway and no activation of other known CTR signaling pathways, including ERK 1/2 and p38 MAP kinases, and Ca 2+ mobilization, supportive of low cell surface receptor expression. Exome sequencing data failed to account for the discrepancy between functional data and expression on the cell lines that do not respond to calcitonin(s) with no deleterious non-synonymous polymorphisms detected, suggesting that other factors may be at play, such as alternative splicing or rapid constitutive receptor internalisation. Conclusions: This study shows that GPCR signaling can display significant variation depending on cellular system used, and effects seen in model recombinant cell lines or tumour cell lines are not always reproduced in a more physiologically relevant system and vice versa.

AB - Background: Glioblastoma (GBM) is the most common and aggressive type of primary brain cancer. With median survival of less than 15 months, identification and validation of new GBM therapeutic targets is of critical importance. Results: In this study we tested expression and performed pharmacological characterization of the calcitonin receptor (CTR) as well as other members of the calcitonin family of receptors in high-grade glioma (HGG) cell lines derived from individual patient tumours, cultured in defined conditions. Previous immunohistochemical data demonstrated CTR expression in GBM biopsies and we were able to confirm CALCR (gene encoding CTR) expression. However, as assessed by cAMP accumulation assay, only one of the studied cell lines expressed functional CTR, while the other cell lines have functional CGRP (CLR/RAMP1) receptors. The only CTR-expressing cell line (SB2b) showed modest coupling to the cAMP pathway and no activation of other known CTR signaling pathways, including ERK 1/2 and p38 MAP kinases, and Ca 2+ mobilization, supportive of low cell surface receptor expression. Exome sequencing data failed to account for the discrepancy between functional data and expression on the cell lines that do not respond to calcitonin(s) with no deleterious non-synonymous polymorphisms detected, suggesting that other factors may be at play, such as alternative splicing or rapid constitutive receptor internalisation. Conclusions: This study shows that GPCR signaling can display significant variation depending on cellular system used, and effects seen in model recombinant cell lines or tumour cell lines are not always reproduced in a more physiologically relevant system and vice versa.

KW - Calcitonin receptor

KW - Glioblastoma

KW - GPCR

KW - Signaling

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U2 - 10.1186/s12885-019-5369-y

DO - 10.1186/s12885-019-5369-y

M3 - Article

VL - 19

JO - BMC Cancer

JF - BMC Cancer

SN - 1471-2407

IS - 1

M1 - 157

ER -