Epicypher, Inc. — Department of Health and Human Services SBIR Phase I: 102

Description

Project Summary Nucleosomes are the fundamental and repeating units of chromatin consisting of DNA wrapped around a histone octamer Alterations in chromatin structure and function dramatically impact downstream gene expression and cellular physiology This epigenetic regulation is controlled by two major modifications histone post translational modifications PTMs such as histone methylation acetylation and ubiquitination and DNA methylation Significantly specific alterations in the epigenetic landscape are associated with a diverse array of human diseases most notable of which are cancers Understanding this so called histone code is essential for developing targeted therapies that manipulate or alter epigenetic signaling to treat human disease As a key step toward deciphering the histone code EpiCypher Inc has initiated a robust research program to develop recombinant nucleosome based reagents and assays which capitalize on physiological properties of chromatin architecture Recent studies show that histone and DNA methylation signaling pathways are interdependent and may synergistically alter the activity of chromatin modifying enzymes These data not only support the hypothesis that histone and DNA methylation dependent signaling are mechanistically linked but provide a unique opportunity for novel drug discovery Currently there are no commercially available research tools to biochemically profile the interplay between specific histone PTMs and DNA methylation on a single assay substrate Here we will develop an innovative nucleosome based assay platform engineered to leverage the crosstalk between histone PTMs DNA methylation and chromatin interacting modifying enzymes to accelerate drug discovery In Phase I we will develop methods to generate recombinant nucleosomes carrying unique DNA and histone methylation profiles Using these reagents as biochemical substrates we will demonstrate feasibility that this assay platform will be useful for drug discovery by establishing novel effector binding and enzymatic assays using proteins known to interact with DNA and or histone methylation such as UHRF UHRF is an E ubiquitin protein ligase whose activity is greatly enhanced in the presence of hemimethylated DNA and histone H K me Significantly UHRF overactivation is associated with bladder and colon cancer In Phase we will continue to optimize the commercial synthesis of designer nucleosomes assembled using methylated DNA templates which we will use to develop additional novel effector binding and enzymatic assays These reagents and assays will be assembled into a series of stand alone kits for innovative drug discovery In addition we will develop a designer nucleosome based UHRF inhibitor screen that leverages the molecular interactions between UHRF and DNA histone methylation to identify context specific inhibitors of UHRF for cancer therapeutic development The breakthrough technology developed here is greatly needed by the epigenetic research community and will enable the identification of novel drug targets as well as the development of precision therapeutics Project Narrative Histone and DNA methylation signaling pathways are interdependent and may synergistically alter the activity of chromatin modifying enzymes However disease relevant interactions between these major epigenetic signaling pathways are difficult to study using currently available research tools Here EpiCypher will develop an innovative recombinant nucleosome based drug discovery platform that leverages interactions between histone and DNA methylation on a single assay substrate The innovative assay platform developed herein will enable the discovery of novel drug targets as well as prove useful in the identification of context specific inhibitors for targeted cancer therapy