An Innovative Non-Hormonal Strategy Targeting Redox Active Metals to Down-Regulate Estrogen-, Progesterone-, Androgen- and Prolactin-Receptors in Breast Cancer

Shehadeh-Tout, F.; Milioli, H. H.; Roslan, S.; Jansson, P. J.; Dharmasivam, M.; Graham, D.; Anderson, R.; Wijesinghe, T.; Azad, M. G.; Richardson, D. R.; Kovacevic, Z.

doi:10.1101/2023.02.02.526543

bioRxiv · 10.1101/2023.02.02.526543

An Innovative Non-Hormonal Strategy Targeting Redox Active Metals to Down-Regulate Estrogen-, Progesterone-, Androgen- and Prolactin-Receptors in Breast Cancer

Shehadeh-Tout, F.·Milioli, H. H.·Roslan, S.·Jansson, P. J.·Dharmasivam, M.·Graham, D.·Anderson, R.·Wijesinghe, T.·Azad, M. G.·Richardson, D. R.·Kovacevic, Z.

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Abstract

Estrogen receptor- (ER-) is a key driver of breast cancer (BC) targeted by tamoxifen. However, tamoxifen resistance is a major problem. An important mechanism of resistance is the activation of EGFR/HER2/HER3 signaling and other hormone receptors (androgen receptor (AR), progesterone receptor (PR), prolactin receptor (PRL-R)) that intrinsically activate ER-. Hence, therapeutics targeting multiple receptors, rather than ER- alone, would be extremely useful and may overcome tamoxifen resistance. This study examined the activity of redox-active di-2-pyridylketone-4,4-dimethyl-3-thiosemicarbazone (Dp44mT) and di-2-pyridylketone-4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC), on the expression and activation of crucial hormone receptors, their co-factors, and key resistance pathways in ER--positive BC. Strikingly, DpC differentially regulated 106 estrogen-response genes with Sankey diagram analysis demonstrating this was linked to decreased mRNA levels of 4 central hormone receptors involved in BC pathogenesis, namely ER, PR, AR, and PRL-R. Mechanistic dissection demonstrated that due to DpC and Dp44mT binding metal ions, these agents caused a pronounced decrease in ER-, AR, PR, and PRL-R protein expression. Ablation of the metal-binding site in the thiosemicarbazone totally prevented its suppressive activity, demonstrating a unique non-hormonal mechanism. DpC and Dp44mT also inhibited EGFR, HER2, and HER3 activation, their downstream signaling, and the expression of co-factors that promote ER- transcriptional activity, including SRC3, NF-{kappa}B p65, and SP1. In vivo, DpC was highly tolerable and effectively inhibited ER--positive BC growth. In conclusion, through a bespoke non-hormonal mechanism targeting redox active metals, Dp44mT and DpC disrupt multiple key inter-receptor interactions between PR, AR, PRL-R, and tyrosine kinases that act with ER- to promote BC, constituting an innovative therapeutic approach.

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Shehadeh-Tout, F., Milioli, H. H., Roslan, S., Jansson, P. J., Dharmasivam, M., Graham, D., Anderson, R., Wijesinghe, T., Azad, M. G., Richardson, D. R., Kovacevic, Z.. 2023-02-03. An Innovative Non-Hormonal Strategy Targeting Redox Active Metals to Down-Regulate Estrogen-, Progesterone-, Androgen- and Prolactin-Receptors in Breast Cancer. https://doi.org/10.1101/2023.02.02.526543

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