title: Anti-leukaemic effects induced by APR-246 are dependent on induction of oxidative stress and the NFE2L2/HMOX1 axis that can be targeted by PI3K and mTOR inhibitors in acute myeloid leukaemia cells.
abstract: The small molecule APR-246 (PRIMA-1(MET) ) is a novel drug that restores the activity of mutated and unfolded TP53 protein. However, the mechanisms of action and potential off-target effects are not fully understood. Gene expression profiling in TP53 mutant KMB3 acute myeloid leukaemia (AML) cells showed that genes which protected cells from oxidative stress to be the most up-regulated. APR-246 exposure also induced reactive oxygen species (ROS) formation and depleted glutathione in AML cells. The genes most up-regulated by APR-246, confirmed by quantitative real time polymerase chain reaction, were heme oxygenase-1 (HMOX1, also termed HO-1), SLC7A11 and RIT1. Up-regulation of HMOX1, a key regulator of cellular response to ROS, was independent of TP53 mutational status. NFE2L2 (also termed Nrf2), a master regulator of HMOX1 expression, showed transcriptional up-regulation and nuclear translocation by APR-246. Down-regulation of NFE2L2 by siRNA in AML cells significantly increased the antitumoural effects of APR-246. The PI3K inhibitor wortmannin and the mTOR inhibitor rapamycin inhibited APR-246-induced nuclear translocation of NFE2L2 and counteracted the protective cellular responses to APR-246, resulting in synergistic cell killing together with APR-246. In conclusion, ROS induction is important for antileukaemic activities of APR-246 and inhibiting the protective response of the Nrf-2/HMOX1 axis using PI3K inhibitors, enhances the antileukaemic effects.
authors.affiliation: Haematology Centre and Centre for Haematology and Regenerative Medicine (HERM), Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Dina
authors.initials: D
authors.lastname: Ali
authors.affiliation: Department of Laboratory Medicine, Clinical Research Centre, Karolinska Institutet, Karolinska Hospital Huddinge, Stockholm, Sweden.
authors.forename: Dara K
authors.initials: DK
authors.lastname: Mohammad
authors.affiliation: Haematology Centre and Centre for Haematology and Regenerative Medicine (HERM), Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Huthayfa
authors.initials: H
authors.lastname: Mujahed
authors.affiliation: Department of Clinical Immunology, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Jonson-Videsäter
authors.affiliation: Department of Laboratory Medicine, Clinical Research Centre, Karolinska Institutet, Karolinska Hospital Huddinge, Stockholm, Sweden.
authors.forename: Beston
authors.initials: B
authors.lastname: Nore
authors.affiliation: Haematology Centre and Centre for Haematology and Regenerative Medicine (HERM), Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Christer
authors.initials: C
authors.lastname: Paul
authors.affiliation: Haematology Centre and Centre for Haematology and Regenerative Medicine (HERM), Karolinska University Hospital, Stockholm, Sweden.|Department of Medical Sciences, Uppsala University, Uppsala, Sweden.
authors.forename: Sören
authors.initials: S
authors.lastname: Lehmann
mesh_terms: D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects*; D051547:Heme Oxygenase-1 / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects* / Q000235:genetics; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000494:pharmacology / Q000627:therapeutic use; D011812:Quinuclidines / Q000494:pharmacology*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; D011812:Quinuclidines; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D051547:Heme Oxygenase-1; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; C533410:eprenetapopt
keywords: APR-246 (PRIMA-1MET); NFE2L2; TP53; acute myeloid leukaemia; reactive oxygen species