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Breast Cancer Research and Treatment

, Volume 146, Issue 2, pp 259–272 | Cite as

A class I histone deacetylase inhibitor, entinostat, enhances lapatinib efficacy in HER2-overexpressing breast cancer cells through FOXO3-mediated Bim1 expression

  • Jangsoon Lee
  • Chandra Bartholomeusz
  • Oula Mansour
  • Juliane Humphries
  • Gabriel N. Hortobagyi
  • Peter Ordentlich
  • Naoto T. Ueno
Preclinical study

Abstract

Although there are effective HER2-targeted agents, novel combination strategies in HER2-overexpressing breast cancers are needed for patients whose tumors develop drug resistance. To develop new therapeutic strategy, we investigated the combinational effect of entinostat, an oral isoform-selective histone deacetylase type I inhibitor, and lapatinib, a HER2/EGFR dual tyrosine kinase inhibitor, in HER2+ breast cancer cells. We assessed the combinational synergistic effect and its mechanism by CellTiter Blue assay, flow cytometry, anchorage-independent growth, quantitative real-time PCR, small interfering RNA, Western blotting, and mammary fat pad xenograft mouse models. We found that compared with entinostat or lapatinib alone, the two drugs in combination synergistically inhibited proliferation (P < 0.001), reduced in vitro colony formation (P < 0.05), and resulted in significant in vivo tumor shrinkage or growth inhibition in two xenograft mouse models (BT474 and SUM190, P < 0.001). The synergistic anti-tumor activity of the entinostat/lapatinib combination was due to downregulation of phosphorylated Akt, which activated transcriptional activity of FOXO3, resulting in induction of Bim1 (a BH3 domain-containing pro-apoptotic protein). Furthermore, entinostat sensitized trastuzumab/lapatinib-resistance-HER2-overexpressing cells to the trastuzumab/lapatinib combination and enhanced the anti-proliferation effect compare with single or double combination treatment. This study provides evidence that entinostat has enhanced anti-tumor effect in combination with HER2-targeted reagent, lapatinib, and resulting in induction of apoptosis by FOXO3-mediated Bim1 expression. Our finding justifies for conducting a clinical trial of combinational treatment with entinostat, lapatinib, and trastuzumab in patients with HER2-overexpressing breast cancer resistant to trastuzumab-based treatment.

Keywords

Entinostat Lapatinib HER2-overexpressing breast cancer FOXO3 Bim1 

Notes

Acknowledgments

This study was supported by the Morgan Welch Inflammatory Breast Cancer Research Program (NTU), the State of Texas Rare and Aggressive Breast Cancer Research Program, and the National Institutes of Health/National Cancer Institute [through CA123318 (NTU) and MD Anderson’s Cancer Center Support Grant, P30CA016672]. We thank Dr. Mien-Chie Hung (MD Anderson Cancer Center) for providing AKT-CA plasmid constructs. We thank Sunita Patterson (Department of Scientific Publications, MD Anderson) for editorial assistance and the Flow Cytometry and Cellular Imaging Facility at MD Anderson for assistance with cell-cycle analysis.

Conflict of interest

Peter Ordentlich is an employee of Syndax Pharmaceuticals. All other authors have no conflicts of interest to disclose.

Supplementary material

10549_2014_3014_MOESM1_ESM.doc (1.9 mb)
Supplementary material 1 (DOC 1909 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Jangsoon Lee
    • 1
  • Chandra Bartholomeusz
    • 1
  • Oula Mansour
    • 1
  • Juliane Humphries
    • 1
  • Gabriel N. Hortobagyi
    • 1
  • Peter Ordentlich
    • 2
  • Naoto T. Ueno
    • 1
  1. 1.Section of Translational Breast Cancer Research, Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, Department of Breast Medical Oncology, Unit 1354The University of Texas MD Anderson Cancer CenterHoustonUSA
  2. 2.Syndax Pharmaceuticals, IncWalthamUSA

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