title: A regimen combining the Wee1 inhibitor AZD1775 with HDAC inhibitors targets human acute myeloid leukemia cells harboring various genetic mutations.
abstract: AZD1775 targets the cell cycle checkpoint kinase Wee1 and potentiates genotoxic agent cytotoxicity through p53-dependent or -independent mechanisms. Here, we report that AZD1775 interacted synergistically with histone deacetylase inhibitors (HDACIs, for example, Vorinostat), which interrupt the DNA damage response, to kill p53-wild type (wt) or -deficient as well as FLT3-ITD leukemia cells in association with pronounced Wee1 inhibition and diminished cdc2/Cdk1 Y15 phosphorylation. Similarly, Wee1 shRNA knockdown significantly sensitized cells to HDACIs. Although AZD1775 induced Chk1 activation, reflected by markedly increased Chk1 S296/S317/S345 phosphorylation leading to inhibitory T14 phosphorylation of cdc2/Cdk1, these compensatory responses were sharply abrogated by HDACIs. This was accompanied by premature mitotic entry, multiple mitotic abnormalities and accumulation of early S-phase cells displaying increased newly replicated DNA, culminating in robust DNA damage and apoptosis. The regimen was active against patient-derived acute myelogenous leukemia (AML) cells harboring either wt or mutant p53 and various next-generation sequencing-defined mutations. Primitive CD34(+)/CD123(+)/CD38(-) populations enriched for leukemia-initiating progenitors, but not normal CD34(+) hematopoietic cells, were highly susceptible to this regimen. Finally, combining AZD1775 with Vorinostat in AML murine xenografts significantly reduced tumor burden and prolonged animal survival. A strategy combining Wee1 with HDACI inhibition warrants further investigation in AML with poor prognostic genetic aberrations.
authors.affiliation: Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: L
authors.initials: L
authors.lastname: Zhou
authors.affiliation: 1] Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA [2] National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun, Jilin, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Chen
authors.affiliation: Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: M
authors.initials: M
authors.lastname: Kmieciak
authors.affiliation: 1] Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA [2] Department of Hematology, Beijing Chaoyang Hospital of Capital Medical University, Beijing, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Leng
authors.affiliation: Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: H
authors.initials: H
authors.lastname: Lin
authors.affiliation: Division of Molecular Diagnostics, Department of Pathology, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: K A
authors.initials: KA
authors.lastname: Rizzo
authors.affiliation: Division of Molecular Diagnostics, Department of Pathology, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: C I
authors.initials: CI
authors.lastname: Dumur
authors.affiliation: Division of Molecular Diagnostics, Department of Pathology, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: A
authors.initials: A
authors.lastname: Ferreira-Gonzalez
authors.affiliation: Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Dai
authors.affiliation: 1] Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA [2] Department of Biochemistry, Virginia Commonwealth University and the Massey Cancer Center and Virginia Institute of Molecular Medicine, Richmond, VA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Grant
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016203:CDC2 Protein Kinase; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000071877:Checkpoint Kinase 1; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D004357:Drug Synergism; D004359:Drug Therapy, Combination; D015973:Gene Expression Regulation, Leukemic; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000235:genetics / Q000401:mortality / Q000473:pathology; D051379:Mice; D022423:Myeloid Cells / Q000378:metabolism / Q000473:pathology; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D061251:Primary Cell Culture; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D016019:Survival Analysis; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D000077337:Vorinostat; D023041:Xenograft Model Antitumor Assays; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D016159:Tumor Suppressor Protein p53; D000077337:Vorinostat; D011494:Protein Kinases; C497970:FLT3 protein, human; D011505:Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3; C499349:WEE1 protein, human; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; C000605869:Chek1 protein, mouse; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; D018844:Cyclin-Dependent Kinases