Epigenetic regulation of microRNA (miRNA) expression has recently been implicated in the pathogenesis of MDS1,2. Particular interest has focused on miRNA124 expression, inhibited in MDS due to promoter hypermethylation and recently demonstrated to be upregulated in response to single and combined epigenetic treatment (EGT)3,4. These observations suggest miRNA124 is implicated in the therapeutic molecular response to EGT and is a potential biomarker of early EGT response in MDS.
Our ongoing studies aimed to evaluate the in vitro and in vivo effects of EGT on known targets of miRNA124 expression including CDK4, CDK6 and EZH2 in order to further characterise the molecular mechanisms associated with early up regulation of miRNA-124 expression and response to EGT.
Analysis of CDK4, CDK6 and EZH2 mRNA expression in HL-60 leukemia cells treated with either the demethylating agent azacytidine (AZA) or the histone deacetylase inhibitor (HDACi) LBH-589 or a combination of these agents demonstrated significant inhibition of CDK4, CDK6 and EZH2 expression. CDK4, CDK6 and EZH2 mRNA expression was also analysed in peripheral blood mononuclear cell samples from high risk MDS/AML patients treated with the combination of AZA and LBH5895. Early and significant in vivo inhibition of CDK4 and CDK6 mRNA expression and a trend to inhibition of EZH2 was identified in patient samples and was also significantly correlated with subsequent clinical and laboratory response to EGT.
Together these observations suggest that in vitro and in vivo expression of CDK4, CDK6 and potentially EZH2, the downstream targets of EGT-induced miRNA124, are inhibited in response to EGT providing a potential molecular mechanism for the miRNA124-mediated response to EGT therapy in patients receiving combination treatment with a demethylating agent and HDACi for high risk MDS/AML.