MI-219-Zinc Combination: a new Paradigm in MDM2 Inhibitor Based Therapy

Zinc plays a crucial role in the biology of p53 in that p53 binds to DNA through a structurally complex domain stabilized by zinc atom. The p53 negative regulator MDM2 protein also carries a C-terminal RING domain that coordinates two zinc atoms which are responsible for p53 nuclear export and proteasomal degradation. In this clinically translatable study, we explored the critical role of zinc on p53 re-activation by MDM2-inhibitor MI-219 in colon and breast cancer cells. ZnCl(2) enhanced MI-219 activity (MTT, apoptosis and colony formation), and chelation of zinc not only blocked the activity of MI-219, it also suppressed re-activation of the p53 and its downstream effector molecules p21(WAF1) and Bax. TPEN, a specific zinc chelator but not Bapta-AM, a calcium chelator, blocked MI-219-induced apoptosis. Nuclear localization is a pre-requisite for proper functioning of p53 and our results confirm that TPEN and not Bapta-AM could abrogate p53 nuclear localization and interfered with p53 transcriptional activation. Addition of zinc suppressed the known p53 feedback MDM2 activation which could be restored by TPEN. Co-immunoprecipitation studies verified that MI-219-mediated MDM2-p53 disruption could be suppressed by TPEN and restored by zinc. As such, single agent therapies that target MDM2 inhibition, without supplemental zinc, may not be optimal in certain patients due to the less recognized mild zinc deficiency among the “at risk population” as in the elderly which are more prone to cancers. Therefore, use of supplemental zinc with MI-219 will benefit the overall efficacy of MDM2 inhibitors and this potent combination warrants further investigation.