Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling
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...
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| Format: | Article |
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Public Library of Science
2018
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| Online Access: | https://eprints.nottingham.ac.uk/48961/ |
| _version_ | 1848797889286897664 |
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| author | Grundy, Martin Seedhouse, Claire Jones, Thomas Elmi, Liban Hall, Michael Graham, Adam Russell, Nigel Pallis, Monica |
| author_facet | Grundy, Martin Seedhouse, Claire Jones, Thomas Elmi, Liban Hall, Michael Graham, Adam Russell, Nigel Pallis, Monica |
| author_sort | Grundy, Martin |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T20:11:03Z |
| format | Article |
| id | nottingham-48961 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T20:11:03Z |
| publishDate | 2018 |
| publisher | Public Library of Science |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-489612020-05-04T19:25:14Z https://eprints.nottingham.ac.uk/48961/ 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 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. Public Library of Science 2018-01-03 Article PeerReviewed Grundy, Martin, Seedhouse, Claire, Jones, Thomas, Elmi, Liban, Hall, Michael, Graham, Adam, Russell, Nigel and Pallis, Monica (2018) Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling. PLoS ONE, 13 (1). e0190682/1-e0190682/19. ISSN 1932-6203 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0190682 doi:10.1371/journal.pone.0190682 doi:10.1371/journal.pone.0190682 |
| spellingShingle | Grundy, Martin Seedhouse, Claire Jones, Thomas Elmi, Liban Hall, Michael Graham, Adam Russell, Nigel Pallis, Monica Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling |
| title | Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling |
| title_full | Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling |
| title_fullStr | Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling |
| title_full_unstemmed | Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling |
| title_short | Predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic BH3 profiling |
| title_sort | predicting effective pro-apoptotic antileukaemic drug combinations using cooperative dynamic bh3 profiling |
| url | https://eprints.nottingham.ac.uk/48961/ https://eprints.nottingham.ac.uk/48961/ https://eprints.nottingham.ac.uk/48961/ |