d,l-Methadone does not improve radio- and chemotherapy in glioblastoma in vitro

Oppermann, Henry; Matusova, Martina; Glasow, Annegret; Dietterle, Johannes; Baran-Schmidt, Rainer; Neumann, Karsten; Meixensberger, Jürgen; Gaunitz, Frank

doi:10.1007/s00280-019-03816-3

Cited by 17 publications

(12 citation statements)

References 25 publications

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“…The authors found no interaction of MTD with TMZ [ 24 ]. Similar observations were made in an independent study with glioblastoma cells and MTD administered together with TMZ or IR (4 Gy) [ 25 ]. It should be noted that in this study TMZ was used at high dose of 200 µM, which causes mainly cytotoxic N-alkylations in the DNA.…”

Section: Discussionsupporting

confidence: 86%

Cytotoxic and Senolytic Effects of Methadone in Combination with Temozolomide in Glioblastoma Cells

Kaina

Beltzig

Piée-Staffa

et al. 2020

IJMS

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Methadone is an analgesic drug used for pain treatment and heroin substitution. Recently, methadone has been proposed to be useful also for cancer therapy, including glioblastoma multiforme (GBM), the most severe form of brain cancer, because experiments on cultured glioma cells treated with doxorubicin showed promising results. Doxorubicin, however, is not used first-line in GBM therapy. Therefore, we analyzed the cytotoxic effect of methadone alone and in combination with temozolomide, a DNA-alkylating drug that is first-line used in GBM treatment, utilizing GBM-derived cell lines and a human fibroblast cell line. We show that methadone is cytotoxic on its own, inducing apoptosis and necrosis, which was observed at a concentration above 20 µg/mL. Methadone was similar toxic in isogenic MGMT expressing and non-expressing cells, and in LN229 glioblastoma and VH10T human fibroblasts. The apoptosis-inducing activity of methadone is not bound on the opioid receptor (OR), since naloxone, a competitive inhibitor of OR, did not attenuate methadone-induced apoptosis/necrosis. Administrating methadone and temozolomide together, temozolomide had no impact on methadone-induced apoptosis (which occurred 3 days after treatment), while temozolomide-induced apoptosis (which occurred 5 days after treatment) was unaffected at low (non-toxic) methadone concentration (5 µg/mL), and at high (toxic) methadone concentration (20 µg/mL) the cytotoxic effects of methadone and temozolomide were additive. Methadone is not genotoxic, as revealed by comet and γH2AX assay, and did not ameliorate the genotoxic effect of temozolomide. Further, methadone did not induce cellular senescence and had no effect on temozolomide-induced senescence. Although methadone was toxic on senescent cells, it cannot be considered a senolytic drug since cytotoxicity was not specific for senescent cells. Finally, we show that methadone had no impact on the MGMT promoter methylation. Overall, the data show that methadone on glioblastoma cells in vitro is cytotoxic and induces apoptosis/necrosis at doses that are above the level that can be achieved in vivo. It is not genotoxic, and does not ameliorate the cell killing or the senescence-inducing effect of temozolomide (no synergistic effect), indicating it has no impact on temozolomide-induced signaling pathways. The data do not support the notion that concomitant methadone treatment supports temozolomide-based chemotherapy.

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Section: Discussionsupporting

confidence: 86%

Cytotoxic and Senolytic Effects of Methadone in Combination with Temozolomide in Glioblastoma Cells

Kaina

Beltzig

Piée-Staffa

et al. 2020

IJMS

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“…One study has already reported no effect of MET on TMZ-induced cytotoxicity in various GBM cell lines [15]. Similar observations were made in another study with GBM cells and MET applied together with TMZ or irradiation (4 Gy) [16]. We could con rm these ndings and most importantly in U373 and A172 cells, MET even reduced sensitivity towards TMZ.…”

Section: Discussionsupporting

confidence: 87%

“…Despite lack of preclinical and clinical data on any bene cial effects of MET on TMZ therapy, it has been promoted as promising therapeutic option for GBM treatment. Current in vitro studies did not report a potentiation of TMZ effects in GBM cells [15,16], questioning the use of MET in GBM (reviewed in [17]).…”

Section: Introductionmentioning

confidence: 99%

Chemotherapy Sensitizing Effects of Methadone in Glioblastoma Cells Are Drug- and Cell Line-dependent

Haas

Ciftcioglu

Jermar

et al. 2020

Preprint

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Background D,L-methadone (MET), an analgesic drug used for pain treatment and opiate addiction has achieved attention from oncologist and social media as possible chemosensitizing agent in glioblastoma multiforme (GBM) treatment. MET has been reported to enhance doxorubicin-induced cytotoxicity in GBM cells via activation of the µ-opioid receptor (MOR) and subsequent apoptosis induction. Here, we further aimed at quantifying MET effects in comparison to other opioids alone and in combination with doxorubicin and clinically more relevant temozolomide (TMZ) in a set of GBM cell lines and primary GBM cells. Methods MOR expression in GBM cells was investigated by immunofluorescence and immunoblotting. Resistance to drugs alone or in combination was assessed by MTT assays. Concentration effect curves were fitted to data points by nonlinear regression analysis and IC50 values were calculated. Apoptotic rates were determined by Annexin V staining. Results We found that MET alone was cytotoxic to GBM cells at high micromolar concentrations in MTT assays by induction of apoptosis and necrosis while morphine and oxycodone were hardly cytotoxic. Naloxone was not able to block MET-induced cytotoxicity, indicating that cell death inducing effects of MET are not MOR dependent. We recorded doxorubicin and TMZ concentration response curves by MTT assays in combination with fixed MET concentrations. MET only enhanced doxorubicin cytotoxicity in one cell line and in part in primary cells at certain MET concentrations. MET was not effective in sensitizing cells towards TMZ. Contrarily, in two cell lines MET even decreased sensitivity towards TMZ. Conclusions MET can be considered cytotoxic to GBM cells only at clinically not relevant concentrations by induction of apoptosis and necrosis. Sensitizing effects are only observed in combination with doxorubicin but not with TMZ and are highly dependent on cell line and applied drug concentrations.

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“…Methadone alone impaired viability (or induced cell death) in A172 [ 16 , 87 ] and U118MG cells [ 16 ] or—in further studies—in U251, U87MG and primary human glioblastoma cells [ 81 , 91 ] as well as in neuroblastoma cells [ 83 ] in vitro with very poor efficacies (IC 50 in the range of ~10 µM–>> 145 µM). Conflictingly, Shi et al, who could not demonstrate a significant apoptosis induction (as defined by annexin-V binding in flow cytometry) with 32 µM (10 µg/mL) methadone in a previous study [ 87 ], more recently reported an induction of annexin-V binding in A172 cells by 0.065 µM methadone [ 82 ].…”

Section: Evidence For a Tumoricidal Activity Of Methadonementioning

confidence: 99%

“…To analyze whether methadone may also enhance the efficacy of the standard anti-glioblastoma therapy [ 9 ], U87MG, U251 and primary human glioblastoma cells were treated with a combination of methadone (1–145 µM) and/or temozolomide (100–200 µM) and/or radiation (4 Gy), and cell survival/death was analyzed by crystal violet staining, annexin-V binding, dehydrogenase activity or ATP measurement. As a result, clinically achievable methadone concentrations are not able to sensitize to temozolomide (apparent methadone IC 50 >>30–>>145 µM) [ 81 , 91 ], to radiation (apparent methadone IC 50 >>30µM) or to concomitant temozolomide-radiation therapy (apparent methadone IC 50 >>30 µM) [ 91 ].…”

Section: Evidence For a Tumoricidal Activity Of Methadonementioning

confidence: 99%

Against Repurposing Methadone for Glioblastoma Therapy

et al. 2020

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Methadone, which is used as maintenance medication for outpatient treatment of opioid dependence or as an analgesic drug, has been suggested by preclinical in vitro and mouse studies to induce cell death and sensitivity to chemo- or radiotherapy in leukemia, glioblastoma, and carcinoma cells. These data together with episodical public reports on long-term surviving cancer patients who use methadone led to a hype of methadone as an anti-cancer drug in social and public media. However, clinical evidence for a tumoricidal effect of methadone is missing and prospective clinical trials, except in colorectal cancer, are not envisaged because of the limited preclinical data available. The present article reviews the pharmacokinetics, potential molecular targets, as well as the evidence for a tumoricidal effect of methadone in view of the therapeutically achievable doses in the brain. Moreover, it provides original in vitro data showing that methadone at clinically relevant concentrations fails to impair clonogenicity or radioresistance of glioblastoma cells.

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d,l-Methadone does not improve radio- and chemotherapy in glioblastoma in vitro

Cited by 17 publications

References 25 publications

Cytotoxic and Senolytic Effects of Methadone in Combination with Temozolomide in Glioblastoma Cells

Cytotoxic and Senolytic Effects of Methadone in Combination with Temozolomide in Glioblastoma Cells

Chemotherapy Sensitizing Effects of Methadone in Glioblastoma Cells Are Drug- and Cell Line-dependent

Against Repurposing Methadone for Glioblastoma Therapy

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