Abstract
Glioblastoma multiforme (GBM) is the most common and aggressive form of brain cancer, with treatment options often constrained due to inherent resistance of malignant cells to conventional therapy. We investigated the impact of triggering programmed cell death (PCD) by using BH3 mimetic drugs in human GBM cell lines. We demonstrate that co-targeting the pro-survival proteins BCL-XL and MCL-1 was more potent at killing six GBM cell lines compared to conventional therapy with Temozolomide or the bromodomain inhibitor JQ1 in vitro. Enhanced cell killing was observed in U251 and SNB-19 cells in response to dual treatment with TMZ or JQ1 combined with a BCL-XL inhibitor, compared to single agent treatment. This was reflected in abundant cleavage/activation of caspase-3 and cleavage of PARP1, markers of apoptosis. U251 and SNB-19 cells were more readily killed by a combination of BH3 mimetics targeting BCL-XL and MCL-1 as opposed to dual treatment with the BCL-2 inhibitor Venetoclax and a BCL-XL inhibitor. The combined loss of BAX and BAK, the essential executioners of intrinsic apoptosis, rendered U251 and SNB-19 cells refractory to any of the drug combinations tested, demonstrating that apoptosis is responsible for their killing. In an orthotopic mouse model of GBM, we demonstrate that the BCL-XL inhibitor A1331852 can penetrate the brain, with A1331852 detected in both tumour and healthy brain regions. We also investigated the impact of combining small molecule inducers of ferroptosis, erastin and RSL3, with BH3 mimetic drugs. We found that a BCL-XL or an MCL-1 inhibitor potently cooperates with inducers of ferroptosis in killing U251 cells. Overall, these findings demonstrate the potential of dual targeting of distinct PCD signalling pathways in GBM and may guide the utility of BCL-XL inhibitors and inducers of ferroptosis with standard of care treatment for improved therapies for GBM.
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Acknowledgements
The authors thank Drs A Morokoff, K Drummond and M Rosenthal for human glioblastoma derived cell lines and discussions about our work.
Funding
This work was supported by a Programme Grant (GNT1113133) to AS and GL, Research Fellowships (GNT1116937 to AS and GNT1117089 to GL) from the Australian NHMRC, the Leukemia & Lymphoma Society of America (Specialized Centre of Research [SCOR] grant no. 7015-18 to AS and GL), a grant-in-aid from the Cure Cancer Foundation (Australia), Leukaemia Foundation Australia grant (SG), the Lady Tata Memorial Trust (SG) and a grant from Cure Brain Cancer Australia grant (AS). Work in the laboratories of the authors was made possible through Victorian State Government Operational Infrastructure Support (OIS) and Australian Government NHMRC Independent Research Institute Infrastructure Support (IRIIS) Scheme.
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DM designed, supervised, and performed research, analysed data, prepared figures, and wrote the manuscript. AS conceived, designed, supervised research, and wrote the manuscript. ES, AGS, FK, KB, MI, RSC, ZW, MS, RK and SG designed and performed some of the research that led to this work and analysed data. DN synthesised A1331852. MRJ, RK, GL and AIB contributed to discussions of the data. All authors reviewed the manuscript.
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DM, ES., KB, FK, MI, RSC, MRJ, DN, ZW, MXS, RMK, GL, SG and AS are or were employees of WEHI which receives royalties from AbbVie and Genentech from the sale of Venetoclax. GL and AS are collaborating with and have received funding from Servier for work on the development of MCL-1-specific BH3 mimetic drugs for cancer therapy. AIB is a shareholder in Alterity Ltd, Cogstate Ltd and Mesoblast Ltd. He is a paid consultant fee for, and has a profit share interest in Collaborative Medicinal Development Pty Ltd.
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Moujalled, D., Southon, A.G., Saleh, E. et al. BH3 mimetic drugs cooperate with Temozolomide, JQ1 and inducers of ferroptosis in killing glioblastoma multiforme cells. Cell Death Differ 29, 1335–1348 (2022). https://doi.org/10.1038/s41418-022-00977-2
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DOI: https://doi.org/10.1038/s41418-022-00977-2
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