UC Santa Barbara

DNA Methyltransferase 3A (DNMT3A) is an epigenetic enzyme known to drive gene silencing, genomic imprinting, and X-chromosome inactivation in a variety of organisms, including humans. By catalyzing the de novo methylation of DNA at CpG sites, DNMT3A establishes the methylation pattern imperative for transcriptional regulation and cellular differentiation during embryonic development. DNMT3A achieves this through interactions with other cellular partners (ncRNA, nucleosomes, p53, and EZH2) to precisely methylate the human genome. Dysregulation of DNMT3A, whether through overexpression, mutation, or loss of regulatory control, leads to aberrant hypermethylation and contributes to various forms of cancer development. For instance, DNMT3A mutations are found in 20-30% of all acute myeloid leukemia (AML) cases, making DNMT3A the most mutated gene in AML patients. Current FDA-approved mechanism-based inhibitors targeting DNMT3A (Azacitidine and Decitabine) are nucleoside analogs that are highly toxic due to their non-discriminant incorporation into the DNA. Considering that AML typically affects older patients, there is a crucial need for more selective, non-cytotoxic DNMT3A inhibitors to control overexpression and aberrant hypermethylation.In this work, we develop fluorescence anisotropy and AlphaLISA assays for high-throughput screening of small-molecules capable of selectively disrupting DNMT3A interactions with histone-tails, central to chromatin recruitment and enzyme activation. Furthermore, we investigate biologically relevant allosteric inhibition of DNMT3A through RNA-mediated epigenetic regulation.