Nonenzymatically modified mRNA for regulating translation and apoptosis by modulating Cancer epigenetics

Abstract

In this study, we employed imidazole-activated natural or modified guanosine derivatives to extend the 3′ ends of mRNA using a nonenzymatic method beyond 30 poly-A tails. We evaluated their impact on the translation activity in cell studies using three genes: GFP, Luciferase, and Apoptin. The assessments were conducted through cell imaging, fluorescence, luminescence, western blot analysis, and RT-qPCR to evaluate varying apoptosis-mediated EZH2 expression in cancer epigenetics, among the compounds tested GMP-2-amino-IM, 2′O-Me-2-amino-IM, and N7-(2-MePy)-GMP-IM. The sugar-modified 2′O-Me-GMP-2-amino-IM demonstrated the most favorable results as mRNAs treated with this compound exhibited higher expression levels with promising mRNA stability relative to the control mRNA (without any extension) and other tested compounds. Subsequently, we transfected cancer cells with nonenzymatically modified apoptin mRNAs by utilizing the three imidazole-activated guanosine derivatives compounds and monitored the induced apoptosis. These findings suggest that 2′O-Me-2-amino-IM-modified apoptin mRNA could serve as a promising tool for cancer therapy by inducing apoptosis while selectively modulating EZH2 expression, a key regulator in oncogene suppression.