gms | German Medical Science

70. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC)
Joint Meeting mit der Skandinavischen Gesellschaft für Neurochirurgie

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

12.05. - 15.05.2019, Würzburg

Methadone induced cell death in glioblastoma cells compared to normal astrocytes

Methadon induzierter Zelltod in Glioblastom Zellen verglichen mit normalen Astrozyten

Meeting Abstract

  • presenting/speaker Konstantin Brawanski - Medizinische Universität Innsbruck, Klinik für Neurochirurgie, Innsbruck, Austria
  • Annette Lohmeier - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland
  • Christian F. Freyschlag - Medizinische Universität Innsbruck, Klinik für Neurochirurgie, Innsbruck, Austria
  • Claudius Thomé - Medizinische Universität Innsbruck, Klinik für Neurochirurgie, Innsbruck, Austria
  • Alexander Brawanski - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland
  • Martin Proescholdt - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland

Deutsche Gesellschaft für Neurochirurgie. 70. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit der Skandinavischen Gesellschaft für Neurochirurgie. Würzburg, 12.-15.05.2019. Düsseldorf: German Medical Science GMS Publishing House; 2019. DocV198

doi: 10.3205/19dgnc213, urn:nbn:de:0183-19dgnc2136

Published: May 8, 2019

© 2019 Brawanski et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License. See license information at http://creativecommons.org/licenses/by/4.0/.


Outline

Text

Objective: Glioblastoma (GBM) is the most frequent primary brain tumor in the adulthood and carries a dismal prognosis. Recently, D,L-methadone has been put forward as adjuvant treatment in GBM. In order to address bioavailability issues and test for in vivo efficacy, we plan to apply D,L-methadone in an implanted rat GBM model. In preparation of this work, we wanted to analyze three aspects:

1.
The efficacy of D,L-methadone alone and in conjunction with temozolomide (TMZ) in a panel of rat GBM cell lines
2.
The susceptibility of normal rat astrocytes to D,L-methadone in order to establish a potential therapeutic window in tumor cells compared to their non-neoplastic counterpart; and
3.
The expression of µ-opiod receptor as the target of D,L-methadone in all tested cell lines.

Methods: We utilized three different established rat GBM cell lines (9L, F98, S635) suitable for in vivo modelling. Normal rat astrocytes were obtained as a primary culture from Fisher 344 rats harvested on day three postnatally. The μ-opioid receptor expression was tested by Western blot and immunofluorescence staining. Cells were exposed to 5 concentrations of D,L-methadone (0.3, 1.0, 15, 30, 45 µg/ml) with or without TMZ (100 µM). Cell viability was tested by crystal violet assay.

Results: Expression of the μ-opioid receptor was similar in the tested cell lines; in particular, there was no difference between the GBM cells compared to normal astrocytes. All GBM lines displayed reduced cell viability upon treatment with D,L-methadone starting at 15 µg/ml, however the astrocytes were entirely refractory. Regarding to the interaction with TMZ 100 µM, low concentrations of D,L-methadone were antagonistic in 9L (0.3 & 1 µg/ml) and F98 (1 µg/ml), whereas higher concentrations (15–45 µg/ml) were synergistic in 9L und S635. In contrast, the astrocytes did not react significantly neither to TMZ nor to TMZ combined with D,L-methadone at any concentration.

Conclusion: Although the target expression is comparable, we found a significantly higher susceptibility to D,L-methadone in GBM cells compared to normal astrocytes. However, the tumor cell killing started at concentrations which will not be reached in patients with GBM. At clinically achievable concentrations, D,L-methadone was antagonistic to TMZ in two GBM lines suggesting caution regarding its premature use in the management of GBM patients