Abstract
Drug treatment of malignant gliomas is limited by the intrinsic resistance of glioma stem cells (GSCs) to chemotherapy. GSCs isolated from human glioblastoma multiforme (GBM) expressed metabotropic glutamate receptors (mGlu3 receptors). The DNA-alkylating agent, temozolomide, killed GSCs only if mGlu3 receptors were knocked down or pharmacologically inhibited. In contrast, mGlu3 receptor blockade did not affect the action of paclitaxel, etoposide, cis-platinum, and irinotecan. mGlu3 receptor blockade enabled temozolomide toxicity by inhibiting a phosphatidylinositol-3-kinase/nuclear factor-κB pathway that supports the expression of O6-methylguanine-DNA methyltransferase (MGMT), an enzyme that confers resistance against DNA-alkylating agents. In mice implanted with GSCs into the brain, temozolomide combined with mGlu3 receptor blockade substantially reduced tumor growth. Finally, 87 patients with GBM undergoing surgery followed by adjuvant chemotherapy with temozolomide survived for longer time if tumor cells expressed low levels of mGlu3 receptors. In addition, the methylation state of the MGMT gene promoter in tumor extracts influenced survival only in those patients with low expression of mGlu3 receptors in the tumor. These data encourage the use of mGlu3 receptor antagonists as add-on drugs in the treatment of GBM, and suggest that the transcript of mGlu3 receptors should be measured in tumor specimens for a correct prediction of patients’ survival in response to temozolomide treatment.
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Abbreviations
- GSC:
-
glioma stem cell
- GBM:
-
glioblastoma multiforme
- MGMT:
-
O6-methylguanine-DNA methyltransferase
- mGlu:
-
metabotropic glutamate
- MAP:
-
mitogen-activated protein
- PtdIns-3-K:
-
phosphatidylinositol-3-kinase
- cAMP:
-
cyclic adenosine monophosphate
- ERK:
-
extracellular signal-regulated kinase
- EGF:
-
epidermal growth factor
- EGFR:
-
epidermal growth factor receptor
- mTOR:
-
mammalian target of rapamycin
- RT-PCR:
-
reverse transcriptase-polymerase chain reaction
- OS:
-
overall survival
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- bFGF:
-
basic fibroblast growth factor
- Sox-2:
-
sex-determining Y-box 2
- DAPI:
-
4,6-diamidino-2-phenylindole
- GSK3:
-
glycogen synthase kinase-3
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- MTT:
-
3-(4,5-dimethylthioazol-2-yl)-2,5-diphenyl tetrazolium bromide
- LDH:
-
lactate dehydrogenase
- ChIP:
-
chromatin immunoprecipitation
- GFP:
-
green fluorescent protein
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Acknowledgements
mGlu2/3 receptor ligands were kindly provided by Eli Lilly and Company. The expression plasmid for β-catenin was provided by Dr. RT Moon (University of Washington, School of Medicine, Seattle, WA, USA). The pCDNA3/constitutively active Akt plasmid was kindly provided by Professor P Formisano, University of Naples ‘Federico II’, Italy. The pCDNA3/MGMT plasmid was kindly provided by Professor Alexandre Evrard, School of Pharmacy, Department of Toxicology, University of Montpellier I (Montpellier, France).
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Ciceroni, C., Bonelli, M., Mastrantoni, E. et al. Type-3 metabotropic glutamate receptors regulate chemoresistance in glioma stem cells, and their levels are inversely related to survival in patients with malignant gliomas. Cell Death Differ 20, 396–407 (2013). https://doi.org/10.1038/cdd.2012.150
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DOI: https://doi.org/10.1038/cdd.2012.150
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