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The histone deacetylase inhibitor butyroyloxymethyl diethylphosphate (AN-7) protects normal cells against toxicity of anticancer agents while augmenting their anticancer activity

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Summary

The histone deacetylase inhibitor (HDACI) butyroyloxymethyl diethylphosphate (AN-7) has been shown to synergize doxorubicin (Dox) anticancer activity while attenuating its cardiotoxicity. In this study we further explored the selectivity of AN-7’s action in several cancer and normal cells treated with anticancer agents. The cells studied were murine mammary 4T1, human breast T47D and glioblastoma U251 cancer cell lines, neonatal rat cardiomyocytes, cardiofibroblasts and astrocytes, and immortalized cardiomyocyte H9C2 cells. Cell death, ROS production and changes in protein expression were measured and in vivo effects were evaluated in Balb-c mice. AN-7 synergized Dox and anti-HER2 cytotoxicity against mammary carcinoma cells with combination indices of 0.74 and 0.79, respectively, while it protected cardiomyocytes against their toxicity. Additionally AN-7 protected astrocytes from Dox-cytoxicity. Cell-type specific changes in the expression of proteins controlling survival, angiogenesis and inflammation by AN-7 or AN-7+Dox were observed. In mice, the protective effect of AN-7 against Dox cardiotoxicity was associated with a reduction in inflammatory factors. In summary, AN-7 augmented the anticancer activity of Dox and anti-HER2 and attenuated their toxicity against normal cells. AN-7 modulation of c-Myc, thrombospondin-1, lo-FGF-2 and other proteins were cell type specific. The effects of AN-7, Dox and their combination were preserved in vivo indicating the potential benefit of combining AN-7 and Dox for clinical use.

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Abbreviations

HDAC:

histone deacetylase

HDACI:

histone deacetylase inhibitor

HAT:

histone acetyltransferase

BA:

butyric acid

Dox:

doxorubicin

NAC:

N-acetyl–L-cysteine

DCF-DA:

2′,7′-dichlorofluorescencin diacetate

PI:

propidium iodide

MEA:

Median Effect Analysis

CI:

combination index

pH2AX:

phosphorylation of histone H2AX

DSBs:

double-strand breaks

lo-FGF-2:

fibroblast growth factor-2

TSP-1:

thrombospondin-1

HO-1:

heme oxigenase-1

SCF:

stem cell factor

CO:

carbon monoxide

TNF-α:

tumor necrosis factor-alpha

INF-γ:

interferon-gamma

HER2:

Human Epidermal growth factor Receptor 2

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Acknowledgments

We thank Ms. S. Dominitz for editorial assistance. This work was supported in part by grants from The Israel Cancer Association (Grant No 20092001, Israel); Teva Special Funds (Dr. A. Swartz) and The Marcus Center for Pharmaceutical and Medicinal Chemistry at Bar Ilan University, Ramat Gan, Israel. Nataly Tarasenko carried out this work as part of the requirements for a PhD degree from the Department of Hematology, Sackler School of Medicine, Tel-Aviv University, Israel.

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Authors and Affiliations

  1. Felsenstein Medical Research Center, Rabin Medical Center, Sackler Faculty of Medicine, Tel-Aviv University, Beilinson Campus, Petach-Tikva, 49100, Israel

    Nataly Tarasenko, Gania Kessler-Icekson, Pnina Boer, Aida Inbal, Hadassa Schlesinger & Ada Rephaeli

  2. Department of Biochemistry, La Trobe University, Melbourne, Victoria, 3086, Australia

    Don R. Phillips & Suzanne M. Cutts

  3. Chemistry Department, Bar-Ilan University, Ramat-Gan, 52900, Israel

    Abraham Nudelman

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  1. Nataly Tarasenko
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Correspondence to Ada Rephaeli.

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Tarasenko, N., Kessler-Icekson, G., Boer, P. et al. The histone deacetylase inhibitor butyroyloxymethyl diethylphosphate (AN-7) protects normal cells against toxicity of anticancer agents while augmenting their anticancer activity. Invest New Drugs 30, 130–143 (2012). https://doi.org/10.1007/s10637-010-9542-z

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