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Acute myeloid leukemia

Small molecule inhibition of cAMP response element binding protein in human acute myeloid leukemia cells

Leukemia volume 30, pages 2302–2311 (2016)Cite this article

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Abstract

The transcription factor CREB (cAMP Response-Element Binding Protein) is overexpressed in the majority of acute myeloid leukemia (AML) patients, and this is associated with a worse prognosis. Previous work revealed that CREB overexpression augmented AML cell growth, while CREB knockdown disrupted key AML cell functions in vitro. In contrast, CREB knockdown had no effect on long-term hematopoietic stem cell activity in mouse transduction/transplantation assays. Together, these studies position CREB as a promising drug target for AML. To test this concept, a small molecule inhibitor of CREB, XX-650-23, was developed. This molecule blocks a critical interaction between CREB and its required co-activator CBP (CREB Binding Protein), leading to disruption of CREB-driven gene expression. Inhibition of CBP–CREB interaction induced apoptosis and cell-cycle arrest in AML cells, and prolonged survival in vivo in mice injected with human AML cells. XX-650-23 had little toxicity on normal human hematopoietic cells and tissues in mice. To understand the mechanism of XX-650-23, we performed RNA-seq, ChIP-seq and Cytometry Time of Flight with human AML cells. Our results demonstrate that small molecule inhibition of CBP–CREB interaction mostly affects apoptotic, cell-cycle and survival pathways, which may represent a novel approach for AML therapy.

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Acknowledgements

We thank Kai Fu and Jing Lu for analysis of ChIP-seq and RNA-seq data. In vivo serum measurements of XX-650-23 were performed in collaboration with Ludmila Alexandrova, PhD, at the Stanford University Mass Spectrometry (SUMS) core facility. Surface Plasmon Resonance (Biacore) analysis was performed in collaboration with Michael Eckart, PhD, at the Stanford Protein and Nucleic Acid (PAN) facility. In vivo animal imaging was performed in collaboration with the Stanford Small Animal Imaging (SCI3) facility, directed by Timothy Doyle, PhD. This research was supported by NIH R01 HL75826, Maxfield Foundation, SPARK program and Child Health Research Institute Lucile Packard Foundation for Children’s Health, Leukemia and Lymphoma Society of America (SLP-8009-15), Pediatric Cancer Research Foundation, Hyundai Hope on Wheels (#02500CA) (KMS), NIH R01 GM087305 (XX), and by the American Cancer Society Greeley & Seattle Gala/Friends of Rob Kinas Postdoctoral Fellowship, Bear Necessities and Jane C Ventura Charitable Trust, Harvey Cohen Endowment, and the Stanford University Dean’s Post-Doctoral Fellowship program (BM).

Author contributions

BM and HDC designed research, performed research, analyzed data and wrote the paper. KH, RD, BCT, AK, FX, BXL and BXL performed research. GAD, RF, KD performed research and analyzed data. GD, GN, MRB, EML, GN, MP and XX analyzed data. NL collected and provided primary cells from AML patients. KMS designed research, analyzed data and wrote the paper.

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  1. B Mitton and H -D Chae: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pediatrics, Stanford University, Stanford, CA, USA

    B Mitton, H -D Chae, K Hsu, R Dutta, G Aldana-Masangkay, K Davis, B C Tiu, A Kaul, N Lacayo, G Dahl, M R Breese & K M Sakamoto

  2. Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, Los Angeles, CA, USA

    R Ferrari & M Pellegrini

  3. Department of Physiology and Pharmacology, Oregon Health and Sciences University, Portland, OR, USA

    F Xie, B X Li & X Xiao

  4. Department of Biomathematics, University of California, Los Angeles, Los Angeles, CA, USA

    E M Landaw

  5. Department of Microbiology and Immunology, Stanford University, Stanford, CA, USA

    G Nolan

  6. Nanosyn, Inc., Santa Clara, CA, USA

    S Romanov

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Correspondence to K M Sakamoto.

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Competing interests

GN has personal financial interest in the company DVS Sciences, the manufacturer that produced some of the reagents and instrumentation used in this manuscript. The remaining authors declare no conflict of interest.

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Mitton, B., Chae, HD., Hsu, K. et al. Small molecule inhibition of cAMP response element binding protein in human acute myeloid leukemia cells. Leukemia 30, 2302–2311 (2016). https://doi.org/10.1038/leu.2016.139

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