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Journal of Cancer Research and Clinical Oncology

, Volume 144, Issue 8, pp 1475–1485 | Cite as

Effects of sequentially applied single and combined temozolomide, hydroxychloroquine and AT101 treatment in a long-term stimulation glioblastoma in vitro model

  • Vivian Adamski
  • Christina Schmitt
  • Florian Ceynowa
  • Rainer Adelung
  • Ralph Lucius
  • Michael Synowitz
  • Kirsten Hattermann
  • Janka Held-Feindt
Original Article – Cancer Research

Abstract

Purpose

Glioblastoma multiforme (GBM) is a poorly curable disease due to its heterogeneity that enables single cells to survive treatment regimen and initiate tumor regrowth. Although some progress in therapy has been achieved in the last years, the efficient treatment of GBMs is still a clinical challenge. Besides the standard therapeutic drug temozolomide (TMZ), quinoline-based antimalarial drugs such as hydroxychloroquine (HCQ) and BH3 mimetics such as AT101 were considered as possible drugs for GBM therapy.

Methods

We investigated the effects of sequentially applied single and combined TMZ, HCQ and AT101 treatments in a long-term stimulation GBM in vitro model. We performed all investigations in parallel in human astrocytes and two differentially TMZ-responsive human GBM cell lines and adjusted used drug concentrations to known liquor/plasma concentrations in patients. We determined amounts of dead cells and still remaining growth rates and depicted our results in a heatmap-like summary to visualize which sequential long-term treatment schedule seemed to be most promising.

Results

We showed that sequential stimulations yielded higher cytotoxicity and better tumor growth control in comparison to single TMZ treatment. This was especially the case for the sequences TMZ/HCQ and TMZ + AT101/AT101 which was as effective as the non-sequential combination TMZ + AT101. Importantly, those affected both less and more TMZ-responsive glioma cell lines, whilst being less harmful for astrocytes in comparison to single TMZ treatment.

Conclusions

Sequential treatment with mechanistically different acting drugs might be an option to reduce side effects in long-term treatment, for example in local administration approaches.

Keywords

R-(−)-gossypol Alternative drugs Sequential treatment Quinoline-based drugs BH3 mimetics Autophagocytosis 

Abbreviations

CHOP

CCAAT-enhancer-binding protein homologous protein

CI

Combination index

CQ

Chloroquine

DMEM

Dulbecco’s modified Eagle’s medium

DMSO

Dimethyl sulfoxide

ER

Endoplasmatic reticulum

FBS

Fetal bovine serum

GAAD-153

Growth arrest and DNA-damage-inducible protein

GBM

Glioblastoma multiforme

HCQ

Hydroxychloroquine

PARP

Poly-ADP ribose polymerase

TMZ

Temozolomide

QNX

Quinacrine

Notes

Acknowledgements

We thank Fereshteh Ebrahim, Brigitte Rehmke, Judith Becker and Sonja Dahle for expert technical assistance.

Author contributions

JHF, KH conceived and designed the study; VA, CS, JHF and KH performed the experiments and analyzed the data; FC, RA, RL and MS contributed materials and assisted in data analysis; JHF and KH wrote the paper, and all authors revised the manuscript.

Funding

This study was funded by the German Research Foundation (DFG) as part as of the Research Training Group “Materials4Brain” (GRK2154; P3, P7 and P8).

Compliance with ethical standards

Conflict of interest

Author Vivian Adamski declares that she has no conflict of interest. Author Christina Schmitt declares that she has no conflict of interest. Author Florian Ceynowa declares that he has no conflict of interest. Author Rainer Adelung declares that he has no conflict of interest. Author Ralph Lucius declares that he has no conflict of interest. Author Michael Synowitz declares that he has no conflict of interest. Author Kirsten Hattermann declares that she has no conflict of interest. Author Janka Held-Feindt declares that she has no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

432_2018_2680_MOESM1_ESM.tif (677 kb)
Supplementary material 1 Figure 1 Supplement : Treatment schedules for the long-term GBM in vitro model. A. Cells were treated for 3 days with the gold standard therapeutic temozolomide (TMZ), switching on day 3 either to the combined treatment of TMZ plus an alternative drug [shown as TMZ+AT101 or TMZ+hydroxychloroquine(HCQ)], or to the single alternative drug (AT101 or HCQ), respective staying with the TMZ treatment. B. Cells were treated for 3 days with one of the single alternative drugs (AT101 or HCQ), switching on day 3 to either the combined therapy with TMZ, to the gold standard therapeutic TMZ or staying with the single alternative drug. C. Cells were treated for 3 days with the combined treatment of TMZ plus one alternative drug, switching on day 3 to either one of the single drugs TMZ, AT101 or HCQ respective staying with the combined therapy until day 6 (TIF 676 KB)
432_2018_2680_MOESM2_ESM.tif (1.3 mb)
Supplementary material 2 Figure 2 Supplement : Effect of selected (non)-sequential treatments on growth rates of astrocytes and two different GBM cell lines treated according to the treatment schedules (ref. Fig. 1 Supplement). The relative growth rates for day 3 and day 6 of SVGA (A), A172 (B) and LN229 (C) were determined compared to the number of cells at day 0 of the treatment. Growth rates of cell treated with the gold standard TMZ are highlighted with dashed lines. Means are significantly different for *p<0.05, **p<0.01 respective ***p<0.001 (TIF 1339 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of NeurosurgeryUniversity Medical Center Schleswig-Holstein UKSHKielGermany
  2. 2.Department of AnatomyUniversity of KielKielGermany
  3. 3.Institute for Materials ScienceUniversity of KielKielGermany

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