Ependymoma stem cells are highly sensitive to temozolomide in vitro and in orthotopic models

Daniela Meco, Tiziana Servidei, Giuseppe Lamorte, Elena Binda, Vincenzo Arena, Riccardo Riccardi

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background Ependymoma management remains challenging because of the inherent chemoresistance of this tumor. To determine whether ependymoma stem cells (SCs) might contribute to therapy resistance, we investigated the sensitivity of ependymoma SCs to temozolomide and etoposide. Methods The efficacies of the two DNA damaging agents were explored in two ependymoma SC lines in vitro and in vivo models. Results Ependymoma SC lines were highly sensitive to temozolomide and etoposide in vitro, but only temozolomide impaired tumor-initiation properties. Consistently, temozolomide but not etoposide showed significant antitumoral activity on ependymoma SC-driven subcutaneous and orthotopic xenografts by reducing the mitotic fraction. In vitro temozolomide at the EC50 (10 μM) induced accumulation of cells in the G 2/M phase that was unexpectedly accompanied by downregulation of p27 and p21 without modulation of full-length p53 (FLp53). Differentiation-committed ependymoma SCs acquired resistance to temozolomide. Inhibition of proliferation was partly due to apoptosis, that occurred earlier in differentiated cells as compared to neurospheres. The activation of apoptosis correlated with an increase in p53β/γ isoforms without modulation of FLp53 under both serum-free and differentiation-promoting media. Incubation of cells in both conditions with temozolomide resulted in increased glioneuronal differentiation exhibiting elevated glial fibrillary acidic protein, galactosylceramidase, and βIII-tubulin expression compared to untreated controls. O 6-methylguanine DNA methyltransferase (MGMT) transcript levels were very low in SCs, and were increased by treatment and, epigenetically, by differentiation through MGMT promoter unmethylation. Conclusion Ependymoma growth might be impaired by temozolomide through preferential depletion of a less differentiated, more tumorigenic, MGMT-negative cell population with stem-like properties.

Original languageEnglish
Pages (from-to)1067-1077
Number of pages11
JournalNeuro-Oncology
Volume16
Issue number8
DOIs
Publication statusPublished - 2014

Fingerprint

temozolomide
Ependymoma
Stem Cells
Etoposide
Methyltransferases
DNA
Galactosylceramidase
O(6)-Methylguanine-DNA Methyltransferase
Apoptosis
Cell Line
Gastrin-Secreting Cells
In Vitro Techniques
Glial Fibrillary Acidic Protein
Tubulin
Heterografts
Cell Division

Keywords

  • Differentiation
  • Ependymoma stem cells
  • MGMT
  • Orthotopic models
  • Temozolomide

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Clinical Neurology
  • Medicine(all)

Cite this

Ependymoma stem cells are highly sensitive to temozolomide in vitro and in orthotopic models. / Meco, Daniela; Servidei, Tiziana; Lamorte, Giuseppe; Binda, Elena; Arena, Vincenzo; Riccardi, Riccardo.

In: Neuro-Oncology, Vol. 16, No. 8, 2014, p. 1067-1077.

Research output: Contribution to journalArticle

Meco, Daniela ; Servidei, Tiziana ; Lamorte, Giuseppe ; Binda, Elena ; Arena, Vincenzo ; Riccardi, Riccardo. / Ependymoma stem cells are highly sensitive to temozolomide in vitro and in orthotopic models. In: Neuro-Oncology. 2014 ; Vol. 16, No. 8. pp. 1067-1077.
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T1 - Ependymoma stem cells are highly sensitive to temozolomide in vitro and in orthotopic models

AU - Meco, Daniela

AU - Servidei, Tiziana

AU - Lamorte, Giuseppe

AU - Binda, Elena

AU - Arena, Vincenzo

AU - Riccardi, Riccardo

PY - 2014

Y1 - 2014

N2 - Background Ependymoma management remains challenging because of the inherent chemoresistance of this tumor. To determine whether ependymoma stem cells (SCs) might contribute to therapy resistance, we investigated the sensitivity of ependymoma SCs to temozolomide and etoposide. Methods The efficacies of the two DNA damaging agents were explored in two ependymoma SC lines in vitro and in vivo models. Results Ependymoma SC lines were highly sensitive to temozolomide and etoposide in vitro, but only temozolomide impaired tumor-initiation properties. Consistently, temozolomide but not etoposide showed significant antitumoral activity on ependymoma SC-driven subcutaneous and orthotopic xenografts by reducing the mitotic fraction. In vitro temozolomide at the EC50 (10 μM) induced accumulation of cells in the G 2/M phase that was unexpectedly accompanied by downregulation of p27 and p21 without modulation of full-length p53 (FLp53). Differentiation-committed ependymoma SCs acquired resistance to temozolomide. Inhibition of proliferation was partly due to apoptosis, that occurred earlier in differentiated cells as compared to neurospheres. The activation of apoptosis correlated with an increase in p53β/γ isoforms without modulation of FLp53 under both serum-free and differentiation-promoting media. Incubation of cells in both conditions with temozolomide resulted in increased glioneuronal differentiation exhibiting elevated glial fibrillary acidic protein, galactosylceramidase, and βIII-tubulin expression compared to untreated controls. O 6-methylguanine DNA methyltransferase (MGMT) transcript levels were very low in SCs, and were increased by treatment and, epigenetically, by differentiation through MGMT promoter unmethylation. Conclusion Ependymoma growth might be impaired by temozolomide through preferential depletion of a less differentiated, more tumorigenic, MGMT-negative cell population with stem-like properties.

AB - Background Ependymoma management remains challenging because of the inherent chemoresistance of this tumor. To determine whether ependymoma stem cells (SCs) might contribute to therapy resistance, we investigated the sensitivity of ependymoma SCs to temozolomide and etoposide. Methods The efficacies of the two DNA damaging agents were explored in two ependymoma SC lines in vitro and in vivo models. Results Ependymoma SC lines were highly sensitive to temozolomide and etoposide in vitro, but only temozolomide impaired tumor-initiation properties. Consistently, temozolomide but not etoposide showed significant antitumoral activity on ependymoma SC-driven subcutaneous and orthotopic xenografts by reducing the mitotic fraction. In vitro temozolomide at the EC50 (10 μM) induced accumulation of cells in the G 2/M phase that was unexpectedly accompanied by downregulation of p27 and p21 without modulation of full-length p53 (FLp53). Differentiation-committed ependymoma SCs acquired resistance to temozolomide. Inhibition of proliferation was partly due to apoptosis, that occurred earlier in differentiated cells as compared to neurospheres. The activation of apoptosis correlated with an increase in p53β/γ isoforms without modulation of FLp53 under both serum-free and differentiation-promoting media. Incubation of cells in both conditions with temozolomide resulted in increased glioneuronal differentiation exhibiting elevated glial fibrillary acidic protein, galactosylceramidase, and βIII-tubulin expression compared to untreated controls. O 6-methylguanine DNA methyltransferase (MGMT) transcript levels were very low in SCs, and were increased by treatment and, epigenetically, by differentiation through MGMT promoter unmethylation. Conclusion Ependymoma growth might be impaired by temozolomide through preferential depletion of a less differentiated, more tumorigenic, MGMT-negative cell population with stem-like properties.

KW - Differentiation

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KW - MGMT

KW - Orthotopic models

KW - Temozolomide

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