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 combina- tions 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 geno- types, including neurofibromatosis type 1 loss, proto-oncogene B-Raf (BRAF)-KIAA1549 fusion, and BRAFV600E 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 glu- tathione 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, prevent- ing the detoxification of carboplatin and inducing increased DNA damage and apoptosis.
Brad Poore, Ming Yuan, Antje Arnold, Antoinette Price, Jesse Alt, Jeffrey A. Rubens, Barbara S. Slusher, Charles G. Eberhart, and Eric H. Raabe
Department of Pathology (B.P., A.A., M.Y., A.P., E.H.R., C.G.E.), Johns Hopkins Drug Discovery (J.A., B.S.S.), Department of Neurology (B.S.S.), Sidney Kimmel Comprehensive Cancer Center (J.A.R., C.G.E., E.H.R.), and Division of Pediatric Oncology (J.A.R., E.H.R.), Johns Hopkins University School of Medicine, Baltimore, Maryland
Corresponding Author: Eric H. Raabe, MD, PhD, Bloomberg Children’s Center Rm 11379, Johns Hopkins Hospital, 1800 Orleans Street, Baltimore, MD 21287 (firstname.lastname@example.org).