Abstract 1011: Induction of ‘BRCAness’ by treatment with histone deacetylase inhibitors sensitizes human triple negative breast cancer cells to PARP inhibitor and cisplatinum.
Authors: Fiskus Warren C. , Rao Rekha , Das Gupta Soumyasri , Chauhan Lata , Bhalla Kapil N. 
Published: August 2014

During DNA damage response (DDR), stability and activity of ATR and CHK1 is essential for the cell cycle arrest and subsequent DNA repair through BRCA1-regulated homologous recombination (HR). We have previously reported that ATR and CHK1 are hsp90 client proteins and treatment with hsp90 inhibitor sensitizes cancer cells to DNA damage. In the present studies, we determined that treatment with pan-HDAC inhibitors vorinostat (VS) and panobinostat (PS) induced hyper-acetylation and inhibition of chaperone function of nuclear hsp90, leading to proteasomal degradation and depletion of ATR, CHK1 and BRCA1. This led to inhibition of DDR and DNA repair following ionizing radiation (IR) through destabilization of ATR-CHK1 and BRCA1 proteins. Specific siRNA-mediated knockdown of HDAC3 but not of HDAC1 or HDAC2 also induced hyper-acetylation of the nuclear hsp90 and depletion of ATR and CHK1, indicating that among the class I HDACs, HDAC3 is the deacetylase for the nuclear hsp90. We next determined whether, by depleting DDR proteins and BRCA1 and inducing ‘BRCAness’, treatment with VS would sensitize the triple negative breast cancer (TNBC) SUM159PT and MB-231 (lacking BRCA1 mutation) as well as BRCA1 mutant HCC1937 cells to the PARP inhibitor ABT888 (veliparib). Indeed, combined treatment with VS or PS and ABT888 (10 to 20 μM) for 48 hours synergistically induced apoptosis (combination indices (CI) by isobologram analysis being &lt; 1.0) of TNBC cells with (HCC1937) or without BRCA1 mutation (MB-231 and SUM159PT cells). As compared to treatment with each agent alone, co-treatment with VS and ABT888 also induced significantly more DNA strand breaks, as demonstrated by higher γ-H2AX levels. Combined treatment also induced markedly greater tail moment, as determined by the comet assay that evaluates the SYBR® Green-stained DNA tails by fluorescent microscopy. Co-treatment with VS and ABT888 also induced more BH3 domain-only pro-death protein BIM, while knockdown of BIM by shRNA significantly reduced the apoptosis induced by co-treatment with VS and ABT888. It is noteworthy that treatment with VS also sensitized the TNBC cells to cisplatin (2.0 to 10 μM)-induced apoptosis. Moreover, co-treatment with VS and ciplatin synergistically induced apoptosis of TNBC cells (CIs &lt; 1.0). These findings indicate that treatment with pan-HDAC inhibitors VS or PS creates ‘BRCAness’, and in combination with a PARP inhibitor or cisplatin synergistically induces apoptosis in human TNBC cells. These findings support a compelling rationale to confirm whether the combination of VS or PS with ABT888 and cisplatin would be a highly active treatment of human TNBC, irrespective of its expression of mutant BRCA1.

Citation Format: Warren C. Fiskus, Rekha Rao, Soumyasri Das Gupta, Lata Chauhan, Kapil N. Bhalla. Induction of ‘BRCAness’ by treatment with histone deacetylase inhibitors sensitizes human triple negative breast cancer cells to PARP inhibitor and cisplatinum. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1011. doi:10.1158/1538-7445.AM2013-1011