Inhibition of mTORC1 in pediatric low-grade glioma depletes glutathione and therapeutically synergizes with carboplatin

Brad Poore, Ming Yuan, Antje Arnold, Antoinette Price, Jesse Alt, Jeffrey Rubens, Barbara Slusher, Charles G Eberhart, Eric Raabe

Research output: Contribution to journalArticle

Abstract

Background Pediatric low-grade glioma (pLGG) often initially responds to front-line therapies such as carboplatin, but more than 50% of treated tumors eventually progress and require additional therapy. With the discovery that pLGG often contains mammalian target of rapamycin (mTOR) activation, new treatment modalities and combinations are now possible for patients. The purpose of this study was to determine if carboplatin is synergistic with the mTOR complex 1 inhibitor everolimus in pLGG. Methods We treated 4 pLGG cell lines and 1 patient-derived xenograft line representing various pLGG genotypes, including neurofibromatosis type 1 loss, proto-oncogene B-Raf (BRAF)-KIAA1549 fusion, and BRAF V600E mutation, with carboplatin and/or everolimus and performed assays for growth, cell proliferation, and cell death. Immunohistochemistry as well as in vivo and in vitro metabolomics studies were also performed. Results Carboplatin synergized with everolimus in all of our 4 pLGG cell lines (combination index <1 at Fa 0.5). Combination therapy was superior at inhibiting tumor growth in vivo. Combination treatment increased levels of apoptosis as well as gamma-H2AX phosphorylation compared with either agent alone. Everolimus treatment suppressed the conversion of glutamine and glutamate into glutathione both in vitro and in vivo. Exogenous glutathione reversed the effects of carboplatin and everolimus. Conclusions The combination of carboplatin and everolimus was effective at inducing cell death and slowing tumor growth in pLGG models. Everolimus decreased the amount of available glutathione inside the cell, preventing the detoxification of carboplatin and inducing increased DNA damage and apoptosis.

Original languageEnglish (US)
Pages (from-to)252-263
Number of pages12
JournalNeuro-oncology
Volume21
Issue number2
DOIs
StatePublished - Feb 14 2019

Fingerprint

Carboplatin
Glioma
Glutathione
Pediatrics
Cell Death
Therapeutics
Growth
Apoptosis
Cell Line
Neoplasms
Neurofibromatosis 1
Metabolomics
Proto-Oncogenes
Sirolimus
mechanistic target of rapamycin complex 1
Everolimus
Glutamine
Heterografts
DNA Damage
Glutamic Acid

Keywords

  • astrocytoma
  • gamma-H2AX
  • mTORC1
  • pediatric brain tumor
  • RPS6

ASJC Scopus subject areas

  • Oncology
  • Clinical Neurology
  • Cancer Research

Cite this

Inhibition of mTORC1 in pediatric low-grade glioma depletes glutathione and therapeutically synergizes with carboplatin. / Poore, Brad; Yuan, Ming; Arnold, Antje; Price, Antoinette; Alt, Jesse; Rubens, Jeffrey; Slusher, Barbara; Eberhart, Charles G; Raabe, Eric.

In: Neuro-oncology, Vol. 21, No. 2, 14.02.2019, p. 252-263.

Research output: Contribution to journalArticle

@article{80baa9dc73554a7ea8ce3d24d99696b6,
title = "Inhibition of mTORC1 in pediatric low-grade glioma depletes glutathione and therapeutically synergizes with carboplatin",
abstract = "Background Pediatric low-grade glioma (pLGG) often initially responds to front-line therapies such as carboplatin, but more than 50{\%} of treated tumors eventually progress and require additional therapy. With the discovery that pLGG often contains mammalian target of rapamycin (mTOR) activation, new treatment modalities and combinations are now possible for patients. The purpose of this study was to determine if carboplatin is synergistic with the mTOR complex 1 inhibitor everolimus in pLGG. Methods We treated 4 pLGG cell lines and 1 patient-derived xenograft line representing various pLGG genotypes, including neurofibromatosis type 1 loss, proto-oncogene B-Raf (BRAF)-KIAA1549 fusion, and BRAF V600E mutation, with carboplatin and/or everolimus and performed assays for growth, cell proliferation, and cell death. Immunohistochemistry as well as in vivo and in vitro metabolomics studies were also performed. Results Carboplatin synergized with everolimus in all of our 4 pLGG cell lines (combination index <1 at Fa 0.5). Combination therapy was superior at inhibiting tumor growth in vivo. Combination treatment increased levels of apoptosis as well as gamma-H2AX phosphorylation compared with either agent alone. Everolimus treatment suppressed the conversion of glutamine and glutamate into glutathione both in vitro and in vivo. Exogenous glutathione reversed the effects of carboplatin and everolimus. Conclusions The combination of carboplatin and everolimus was effective at inducing cell death and slowing tumor growth in pLGG models. Everolimus decreased the amount of available glutathione inside the cell, preventing the detoxification of carboplatin and inducing increased DNA damage and apoptosis.",
keywords = "astrocytoma, gamma-H2AX, mTORC1, pediatric brain tumor, RPS6",
author = "Brad Poore and Ming Yuan and Antje Arnold and Antoinette Price and Jesse Alt and Jeffrey Rubens and Barbara Slusher and Eberhart, {Charles G} and Eric Raabe",
year = "2019",
month = "2",
day = "14",
doi = "10.1093/neuonc/noy150",
language = "English (US)",
volume = "21",
pages = "252--263",
journal = "Neuro-Oncology",
issn = "1522-8517",
publisher = "Oxford University Press",
number = "2",

}

TY - JOUR

T1 - Inhibition of mTORC1 in pediatric low-grade glioma depletes glutathione and therapeutically synergizes with carboplatin

AU - Poore, Brad

AU - Yuan, Ming

AU - Arnold, Antje

AU - Price, Antoinette

AU - Alt, Jesse

AU - Rubens, Jeffrey

AU - Slusher, Barbara

AU - Eberhart, Charles G

AU - Raabe, Eric

PY - 2019/2/14

Y1 - 2019/2/14

N2 - Background Pediatric low-grade glioma (pLGG) often initially responds to front-line therapies such as carboplatin, but more than 50% of treated tumors eventually progress and require additional therapy. With the discovery that pLGG often contains mammalian target of rapamycin (mTOR) activation, new treatment modalities and combinations are now possible for patients. The purpose of this study was to determine if carboplatin is synergistic with the mTOR complex 1 inhibitor everolimus in pLGG. Methods We treated 4 pLGG cell lines and 1 patient-derived xenograft line representing various pLGG genotypes, including neurofibromatosis type 1 loss, proto-oncogene B-Raf (BRAF)-KIAA1549 fusion, and BRAF V600E mutation, with carboplatin and/or everolimus and performed assays for growth, cell proliferation, and cell death. Immunohistochemistry as well as in vivo and in vitro metabolomics studies were also performed. Results Carboplatin synergized with everolimus in all of our 4 pLGG cell lines (combination index <1 at Fa 0.5). Combination therapy was superior at inhibiting tumor growth in vivo. Combination treatment increased levels of apoptosis as well as gamma-H2AX phosphorylation compared with either agent alone. Everolimus treatment suppressed the conversion of glutamine and glutamate into glutathione both in vitro and in vivo. Exogenous glutathione reversed the effects of carboplatin and everolimus. Conclusions The combination of carboplatin and everolimus was effective at inducing cell death and slowing tumor growth in pLGG models. Everolimus decreased the amount of available glutathione inside the cell, preventing the detoxification of carboplatin and inducing increased DNA damage and apoptosis.

AB - Background Pediatric low-grade glioma (pLGG) often initially responds to front-line therapies such as carboplatin, but more than 50% of treated tumors eventually progress and require additional therapy. With the discovery that pLGG often contains mammalian target of rapamycin (mTOR) activation, new treatment modalities and combinations are now possible for patients. The purpose of this study was to determine if carboplatin is synergistic with the mTOR complex 1 inhibitor everolimus in pLGG. Methods We treated 4 pLGG cell lines and 1 patient-derived xenograft line representing various pLGG genotypes, including neurofibromatosis type 1 loss, proto-oncogene B-Raf (BRAF)-KIAA1549 fusion, and BRAF V600E mutation, with carboplatin and/or everolimus and performed assays for growth, cell proliferation, and cell death. Immunohistochemistry as well as in vivo and in vitro metabolomics studies were also performed. Results Carboplatin synergized with everolimus in all of our 4 pLGG cell lines (combination index <1 at Fa 0.5). Combination therapy was superior at inhibiting tumor growth in vivo. Combination treatment increased levels of apoptosis as well as gamma-H2AX phosphorylation compared with either agent alone. Everolimus treatment suppressed the conversion of glutamine and glutamate into glutathione both in vitro and in vivo. Exogenous glutathione reversed the effects of carboplatin and everolimus. Conclusions The combination of carboplatin and everolimus was effective at inducing cell death and slowing tumor growth in pLGG models. Everolimus decreased the amount of available glutathione inside the cell, preventing the detoxification of carboplatin and inducing increased DNA damage and apoptosis.

KW - astrocytoma

KW - gamma-H2AX

KW - mTORC1

KW - pediatric brain tumor

KW - RPS6

UR - http://www.scopus.com/inward/record.url?scp=85062225327&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85062225327&partnerID=8YFLogxK

U2 - 10.1093/neuonc/noy150

DO - 10.1093/neuonc/noy150

M3 - Article

VL - 21

SP - 252

EP - 263

JO - Neuro-Oncology

JF - Neuro-Oncology

SN - 1522-8517

IS - 2

ER -