Effects of sequentially applied single and combined temozolomide, hydroxychloroquine and AT101 treatment in a long-term stimulation glioblastoma in vitro model
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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.
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.
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.
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.
KeywordsR-(−)-gossypol Alternative drugs Sequential treatment Quinoline-based drugs BH3 mimetics Autophagocytosis
CCAAT-enhancer-binding protein homologous protein
Dulbecco’s modified Eagle’s medium
Fetal bovine serum
Growth arrest and DNA-damage-inducible protein
Poly-ADP ribose polymerase
We thank Fereshteh Ebrahim, Brigitte Rehmke, Judith Becker and Sonja Dahle for expert technical assistance.
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.
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.
This article does not contain any studies with human participants or animals performed by any of the authors.
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