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Acute Leukemias

A regimen combining the Wee1 inhibitor AZD1775 with HDAC inhibitors _targets human acute myeloid leukemia cells harboring various genetic mutations

Leukemia volume 29, pages 807–818 (2015)Cite this article

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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.

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Acknowledgements

This work was supported by the National Institutes of Health (Grant CA93738, CA100866, CA167708, P50 CA142509, and P50 CA130805) and the Leukemia and Lymphoma Society of America (Grant R6238 and 6472-15). Plasmid preparation was performed at the VCU Macromolecule Core Facility, supported, in part, with funding from NIH-NCI Cancer Center Core Grant 5P30CA016059-29; Confocal microscopies were performed at the VCU Department of Anatomy and Neurobiology Microscopy Facility, supported, in part, with funding from NIH-NINDS Center Core Grant 5P30NS047463.

Author Contributions

YD and SG conceptualized research and formed the hypothesis of this paper; SC and YD designed, performed experiments, analyzed data; LZ, YZ, SC, YL, HL and YD performed the in vitro research and collected data; LZ, YZ, SC and YD performed animal studies and collected data; MK designed, performed and analyzed flow cytometry work; KAR, CID and AF-G provided patient samples, or performed and analyzed NGS work; YD and SG wrote the manuscript.

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  1. L Zhou and Y Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Hematology and Oncology, Department of Medicine, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA

    L Zhou, Y Zhang, S Chen, M Kmieciak, Y Leng, H Lin, Y Dai & S Grant

  2. National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun, Jilin, China

    Y Zhang

  3. Department of Hematology, Beijing Chaoyang Hospital of Capital Medical University, Beijing, China

    Y Leng

  4. Division of Molecular Diagnostics, Department of Pathology, Virginia Commonwealth University and the Massey Cancer Center, Richmond, VA, USA

    K A Rizzo, C I Dumur & A Ferreira-Gonzalez

  5. Department of Biochemistry, Virginia Commonwealth University and the Massey Cancer Center and Virginia Institute of Molecular Medicine, Richmond, VA, USA

    S Grant

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Zhou, L., Zhang, Y., Chen, S. et al. A regimen combining the Wee1 inhibitor AZD1775 with HDAC inhibitors _targets human acute myeloid leukemia cells harboring various genetic mutations. Leukemia 29, 807–818 (2015). https://doi.org/10.1038/leu.2014.296

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