Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling

Grundy, Martin; Seedhouse, Claire; Jones, Thomas; Elmi, Liban; Hall, Michael; Graham, Adam; Russell, Nigel; Pallis, Monica

Abstract

The BH3-only apoptosis agonists BAD and NOXA target BCL-2 and MCL-1 respectively and co-operate to induce apoptosis. On this basis, therapeutic drugs targeting BCL-2 and MCL-1 might have enhanced activity if used in combination. We identified anti-leukaemic drugs sensitising to BCL-2 antagonism and drugs sensitising to MCL-1 antagonism using the technique of dynamic BH3 profiling, whereby cells were primed with drugs to discover whether this would elicit mitochondrial outer membrane permeabilisation in response to BCL-2-targeting BAD-BH3 peptide or MCL-1-targeting MS1-BH3 peptide. We found that a broad range of anti-leukaemic agents–notably MCL-1 inhibitors, DNA damaging agents and FLT3 inhibitors–sensitise leukaemia cells to BAD-BH3. We further analysed the BCL-2 inhibitors ABT-199 and JQ1, the MCL-1 inhibitors pladienolide B and torin1, the FLT3 inhibitor AC220 and the DNA double-strand break inducer etoposide to correlate priming responses with co-operative induction of apoptosis. ABT-199 in combination with pladienolide B, torin1, etoposide or AC220 strongly induced apoptosis within 4 hours, but the MCL-1 inhibitors did not co-operate with etoposide or AC220. In keeping with the long half-life of BCL-2, the BET domain inhibitor JQ1 was found to downregulate BCL-2 and to prime cells to respond to MS1-BH3 at 48, but not at 4 hours: prolonged priming with JQ1 was then shown to induce rapid cytochrome C release when pladienolide B, torin1, etoposide or AC220 were added. In conclusion, dynamic BH3 profiling is a useful mechanism-based tool for understanding and predicting co-operative lethality between drugs sensitising to BCL-2 antagonism and drugs sensitising to MCL-1 antagonism. A plethora of agents sensitised cells to BAD-BH3-mediated mitochondrial outer membrane permeabilisation in the dynamic BH3 profiling assay and this was associated with effective co-operation with the BCL-2 inhibitory compounds ABT-199 or JQ1.

Citation

Grundy, M., Seedhouse, C., Jones, T., Elmi, L., Hall, M., Graham, A., …Pallis, M. (2018). Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling. PLoS ONE, 13(1), Article e0190682. https://doi.org/10.1371/journal.pone.0190682

• Journal Article Type: Article
• Acceptance Date: Dec 22, 2017
• Publication Date: Jan 3, 2018
• Deposit Date: Jan 8, 2018
• Publicly Available Date: Jan 8, 2018
• Journal: PLoS ONE
• Electronic ISSN: 1932-6203
• Publisher: Public Library of Science
• Peer Reviewed: Peer Reviewed
• Volume: 13
• Issue: 1
• Article Number: e0190682
• DOI: https://doi.org/10.1371/journal.pone.0190682
• Public URL: https://nottingham-repository.worktribe.com/output/902874
• Publisher URL: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0190682
• Contract Date: Jan 8, 2018