Effects of sequentially applied single and combined temozolomide, hydroxychloroquine and AT101 treatment in a long-term stimulation glioblastoma in vitro model
- 270 Downloads
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.
- Adamski V, Hempelmann A, Flüh C, Lucius R, Synowitz M, Hattermann K, Held-Feindt J (2017) Dormant human glioblastoma cells acquire stem cell characteristics and are differentially affected by temozolomide and AT101 treatment strategies. Oncotarget 8:108064–108078CrossRefPubMedPubMedCentralGoogle Scholar
- Antonietti P, Linder B, Hehlgans S, Mildenberger IC, Burger MC, Fulda S, Steinbach JP, Gessler F, Rödel F, Mittelbronn M, Kögel D (2017) Interference with the HSF1/HSP70/BAG3 pathway primes glioma cells to matrix detachment and BH3 mimetic-induced apoptosis. Mol Cancer Ther 16:156–168CrossRefPubMedGoogle Scholar
- Hsu SPC, Kuo JS, Chiang HC, Wang HE, Wang YS, Huang CC, Huang YC, Chi MS, Mehta MP, Chi KH (2018) Temozolomide, sirolimus and chloroquine is a new therapeutic combination that synergizes to disrupt lysosomal function and cholesterol homeostasis in GBM cells. Oncotarget 9:6883–6896PubMedPubMedCentralGoogle Scholar
- Jarzabek MA, Amberger-Murphy V, Callanan JJ, Gao C, Zagozdzon AM, Shiels L, Wang J, Ligon KL, Rich BE, Dicker P, Gallagher WM, Prehn JH, Byrne AT (2014) Interrogation of gossypol therapy in glioblastoma implementing cell line and patient-derived tumor models. Br J Cancer 11:2275–2286CrossRefGoogle Scholar
- Lee SY (2016) Temozolomide resistance in glioblastoma multiforme. Genes Dis 3:198e210Google Scholar
- Rangwala R, Leone R, Chang YC, Fecher LA, Schuchter LM, Kramer A, Tan KS, Heitjan DF, Rodgers G, Gallagher M, Piao S, Troxel AB, Evans TL, DeMichele AM, Nathanson KL, O’Dwyer PJ, Kaiser J, Pontiggia L, Davis LE, Amaravadi RK (2014) Phase I trial of hydroxychloroquine with dose-intense temozolomide in patients with advanced solid tumors and melanoma. Autophagy 10:1369–1379CrossRefPubMedPubMedCentralGoogle Scholar
- Rosenfeld MR, Ye X, Supko JG, Desideri S, Grossman SA, Brem S, Mikkelson T, Wang D, Chang YC, Hu J, McAfee Q, Fisher J, Troxel AB, Piao S, Heitjan DF, Tan KS, Pontiggia L, O’Dwyer PJ, Davis LE, Amaravadi RK (2014) A phase I/II trial of hydroxychloroquine in conjunction with radiation therapy and concurrent and adjuvant temozolomide in patients with newly diagnosed glioblastoma multiforme. Autophagy 10:1359–1368CrossRefPubMedPubMedCentralGoogle Scholar
- Schelman WR, Mohammed TA, Traynor AM, Kolesar JM, Marnocha RM, Eickhoff J, Keppen M, Alberti DB, Wilding G, Takebe N, Liu G (2014) A phase I study of AT-101 with cisplatin and etoposide in patients with advanced solid tumors with an expanded cohort in extensive-stage small cell lung cancer. Investig New Drugs 32:295–302CrossRefGoogle Scholar
- Shen S, Wu Y, Li K, Wang Y, Wu J, Zeng Y, Wu D (2018) Versatile hyaluronic acid modified AQ4N-Cu(II)-gossypol infinite coordination polymer nanoparticles: multiple tumor targeting, highly efficient synergistic chemotherapy, and real-time self-monitoring. Biomaterials 154:197–212CrossRefPubMedGoogle Scholar
- Sonpavde G, Matveev V, Burke JM, Caton JR, Fleming MT, Hutson TE, Galsky MD, Berry WR, Karlov P, Holmlund JT, Wood BA, Brookes M, Leopold L (2012) Randomized phase II trial of docetaxel plus prednisone in combination with placebo or AT-101, an oral small molecule Bcl-2 family antagonist, as first-line therapy for metastatic castration-resistant prostate cancer. Ann Oncol 23:1803–1808CrossRefPubMedGoogle Scholar
- Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO, European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups, National Cancer Institute of Canada Clinical Trials Group (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:987–996CrossRefPubMedGoogle Scholar
- Wang Y, Shi K, Zhang L, Hu G, Wan J, Tang J, Yin S, Duan J, Qin M, Wang N, Xie D, Gao X, Gao H, Zhang Z, He Q (2016) Significantly enhanced tumor cellular and lysosomal hydroxychloroquine delivery by smart liposomes for optimal autophagy inhibition and improved antitumor efficiency with liposomal doxorubicin. Autophagy 12:949–962CrossRefPubMedPubMedCentralGoogle Scholar
- Zerp SF, Stoter TR, Hoebers FJP, van den Brekel MWM, Dubbelman R, Kuipers GK, Lafleur MVM, Slotman BJ, Verheij M (2015) Targeting anti-apoptotic Bcl-2 by AT-101 to increase radiation efficacy: data from in vitro and clinical pharmacokinetic studies in head and neck cancer. Radiat Oncol 10:158CrossRefPubMedPubMedCentralGoogle Scholar