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Suberoylanilide hydroxamic acid (SAHA) enhances olaparib activity by targeting homologous recombination DNA repair in ovarian cancer

  • Author Footnotes
    1 Contributed equally.
    Panagiotis A. Konstantinopoulos
    Footnotes
    1 Contributed equally.
    Affiliations
    Department of Medical Oncology, Medical Gynecologic Oncology Program, Dana Farber Cancer Institute, Harvard Medical School, Boston, MA, USA
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  • Author Footnotes
    1 Contributed equally.
    Andrew J. Wilson
    Footnotes
    1 Contributed equally.
    Affiliations
    Department of Obstetrics & Gynecology, Division of Gynecologic Oncology, Vanderbilt University Medical Center, Nashville, TN, USA
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  • Jeanette Saskowski
    Affiliations
    Department of Obstetrics & Gynecology, Division of Gynecologic Oncology, Vanderbilt University Medical Center, Nashville, TN, USA
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  • Erica Wass
    Affiliations
    Department of Obstetrics & Gynecology, Division of Gynecologic Oncology, Vanderbilt University Medical Center, Nashville, TN, USA
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  • Dineo Khabele
    Correspondence
    Corresponding author at: Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Vanderbilt University, 21st Avenue South, B-1100 Medical Center North, Nashville, TN 37232, USA. Fax: +1 615 343 8403.
    Affiliations
    Department of Obstetrics & Gynecology, Division of Gynecologic Oncology, Vanderbilt University Medical Center, Nashville, TN, USA

    Vanderbilt-Ingram Cancer Center, Nashville, TN, USA
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  • Author Footnotes
    1 Contributed equally.

      Highlights

      • SAHA downregulates homologous recombination pathway genes in ovarian cancer cells.
      • Formation of RAD51 foci is inhibited by SAHA alone and combined with olaparib.
      • Anti-tumor effects of olaparib in vitro and in vivo are enhanced by SAHA.

      Abstract

      Objectives

      Approximately 50% of serous epithelial ovarian cancers (EOC) contain molecular defects in homologous recombination (HR) DNA repair pathways. Poly(ADP-ribose) polymerase inhibitors (PARPi) have efficacy in HR-deficient, but not in HR-proficient, EOC tumors as a single agent. Our goal was to determine whether the histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), can sensitize HR-proficient ovarian cancer cells to the PARPi AZD-2281 (olaparib).

      Methods

      Ovarian cancer cell lines (SKOV-3, OVCAR-8, NCI/ADR-Res, UWB1.289 BRCA1null and UWB1.289 + BRCA1 wild-type) were treated with saline vehicle, olaparib, SAHA or olaparib/SAHA. Sulforhodamine B (SRB) assessed cytotoxicity and immunofluorescence and Western blot assays assessed markers of apoptosis (cleaved PARP) and DNA damage (pH2AX and RAD51). Drug effects were also tested in SKOV-3 xenografts in Nude mice. Affymetrix microarray experiments were performed in vehicle and SAHA-treated SKOV-3 cells.

      Results

      In a microarray analysis, SAHA induced coordinated down-regulation of HR pathway genes, including RAD51 and BRCA1. Nuclear co-expression of RAD51 and pH2AX, a marker of efficient HR repair, was reduced approximately 40% by SAHA treatment alone and combined with olaparib. SAHA combined with olaparib induced apoptosis and pH2AX expression to a greater extent than either drug alone. Olaparib reduced cell viability at increasing concentrations and SAHA enhanced these effects in 4 of 5 cell lines, including BRCA1 null and wild-type cells, in vitro and in SKOV-3 xenografts in vivo.

      Conclusions

      These results provide preclinical rationale for targeting DNA damage response pathways by combining small molecule PARPi with HDACi as a mechanism for reducing HR efficiency in ovarian cancer.

      Keywords

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