Translational Therapeutics

Reversal of glucocorticoid resistance in paediatric acute lymphoblastic leukaemia is dependent on restoring BIM expression

Abstract

Background

Acute lymphoblastic leukaemia (ALL) is the most common paediatric malignancy. Glucocorticoids form a critical component of chemotherapy regimens and resistance to glucocorticoid therapy is predictive of poor outcome. We have previously shown that glucocorticoid resistance is associated with upregulation of the oncogene C-MYC and failure to induce the proapoptotic gene BIM.

Methods

A high-throughput screening (HTS) campaign was carried out to identify glucocorticoid sensitisers against an ALL xenograft derived from a glucocorticoid-resistant paediatric patient. Gene expression analysis was carried out using Illumina microarrays. Efficacy, messenger RNA and protein analysis were carried out by Resazurin assay, reverse transcription-PCR and immunoblotting, respectively.

Results

A novel glucocorticoid sensitiser, 2-((4,5-dihydro-1H-imidazol-2-yl)thio)-N-isopropyl-N-phenylacetamide (GCS-3), was identified from the HTS campaign. The sensitising effect was specific to glucocorticoids and synergy was observed in a range of dexamethasone-resistant and dexamethasone-sensitive xenografts representative of B-ALL, T-ALL and Philadelphia chromosome-positive ALL. GCS-3 in combination with dexamethasone downregulated C-MYC and significantly upregulated BIM expression in a glucocorticoid-resistant ALL xenograft. The GCS-3/dexamethasone combination significantly increased binding of the glucocorticoid receptor to a novel BIM enhancer, which is associated with glucocorticoid sensitivity.

Conclusions

This study describes the potential of the novel glucocorticoid sensitiser, GCS-3, as a biological tool to interrogate glucocorticoid action and resistance.

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Fig. 1: Ex vivo efficacy of GCS-3 in combination with dexamethasone or prednisolone against ALL-19 xenograft cells.
Fig. 2: Ex vivo efficacy of GCS-3 in combination with dexamethasone against ALL xenograft cells.
Fig. 3: GCS-3 requires a functional GR to induce caspase-dependent apoptosis.
Fig. 4: Effects of GCS-3 on dexamethasone-induced gene expression changes.
Fig. 5: Expression levels of candidate genes in ALL-19 cells.
Fig. 6: BIM upregulation correlates with GCS-3/dexamethasone efficacy.

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Acknowledgements

We thank Professor John E. Pimanda for kindly providing CD34+ cells from human cord blood and Dr. Julie A.I. Thoms for technical support. We also thank donors and staff from the Sydney Cord Blood Bank for supplying cord bloods. Children’s Cancer Institute is affiliated with UNSW Sydney and the Sydney Children’s Hospitals Network.

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C.E.T. and R.B.L. designed the study; C.E.T., D.J. and C.M. generated and analysed the data; C.E.T. and R.B.L. interpreted the data and wrote the manuscript.

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Correspondence to Richard B. Lock.

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All samples used in this manuscript were either purchased, established cell lines or patient-derived xenografts (passage ≥ 2) that had previously been established with appropriate informed consent. All patient-derived xenografts were previously established under approval of the University of New South Wales Animal Care and Ethics Committee. Cord blood was obtained from the Sydney Cord Blood Bank under approval of the Prince of Wales Hospital (Reference number HREC 08/190). All donors consent to their sample being used for research and the Sydney Cord Blood Bank is licensed by the Therapeutic Goods Administration and accredited by the Foundation for Accreditation of Cellular Therapy. The study was performed in accordance with the Declaration of Helsinki.

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This manuscript does not contain any individual person’s data in any form.

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All data generated or analysed during this study are included in this manuscript. Supplementary information is available at the British Journal of Cancer website.

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The authors declare no competing interests.

Funding information

This research was supported through the Priority-driven Collaborative Cancer Research Scheme and co-funded by Cancer Australia and The Kids’ Cancer Project (Grant APP1129129). This research was also funded by the Anthony Rothe Memorial Trust, the National Health and Medical Research Council of Australia (NHMRC Fellowships APP1059804 and APP1157871 to RBL) and the Cancer Institute NSW (Early Career Fellowship 15/ECF/1-02 to D.J.). C.E.T. was funded by a Research Excellence Award from UNSW Sydney.

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Toscan, C.E., Jing, D., Mayoh, C. et al. Reversal of glucocorticoid resistance in paediatric acute lymphoblastic leukaemia is dependent on restoring BIM expression. Br J Cancer (2020). https://doi.org/10.1038/s41416-020-0824-8

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