Accelerated discovery of methylation targeted therapeutic development

PROJECT SUMMARYNucleosomesNucsare the repeating units of chromatinmade up of DNA wrapped around a histone octamerChanges in chromatin structurefunction can dramatically impact downstream gene expression and cellular physiologydriving oncogenic reprogramming and cancer progressionThis epigenetic regulation of chromatin is controlled by two major modificationshistone post translational modificationsPTMssuch as histone methylation or acetylationand DNA methylationRecent studies show that histone and DNA methylation signaling pathways are interdependent and synergistically alter the activity of chromatin modifying interactor proteinsIndeedmost chromatin regulators contain multiple chromatin binding modules that interact with both DNA and histone modificationssuggesting that combinatorial interactionsvia DNA and histonesare fundamental to their regulation and function in vivoHoweverthere are currently no commercially available research tools to biochemically profile the interplay between specific histone and DNA modifications on a single assay substrateHereEpiCypher is developing and commercializing high throughput assays that leverage recombinant designer nucleosomesdNucscontaining both histone PTMs and DNA methylationMeDNA dNucsThis firstto market assay platform is highly innovativeunlocking access to epigenetic drug targets that require combinatorial nucleosome modifications to recapitulate in vivo binding specificityenzyme activitysee UHRFI example belowIn Phase Iwe have developed methods to generate dNucs carrying unique DNA methylation profilesMeDNA dNucsand validated these reagents using effector binding and enzymatic assaysSignificantlywe discovered an optimized MeDNA dNuc substrate for the oncogenic histone targeting ubiquitin ligase enzyme UHRFwhich is overexpressed in many cancers and a powerful indicator of poor prognosisIndeedthe binding preferenceactivityand target selectivity of UHRFis dramatically altered when targeting MeDNA dNucs vsunmodified NucsHerewe will develop additional disease relevant methylated DNA speciesmChmCfCandcaCas we scale up manufacturingreduce costsincrease throughputof a diverse set of MeDNA dNucs for therapeutic development and discoveryAimNextwe will functionally validate our MeDNA dNuc library using EpiCodeTMEpiCypher s high throughput discovery platformAimdemonstrating how these substrates can be used for novel drugdisease biomarker discoveryThis work will be performed in collaboration with DrScott Rothbartan expert in DNAhistone modification crosstalkFinallywe will demonstrate the utility of MeDNA dNucs for drug development by developing and validating the first HTS inhibitor assays for UHRFAimbinding and enzymatic assaysThis platform will provide novel access to challengingor so calledundruggableepigenetic targets as well as help decipher complex chromatin signaling interactions toward the identification of new cancer biomarkers PROJECT NARRATIVEHistone and DNA methylation signaling pathways are interdependent and synergistically alter the binding activity of chromatin interactor proteinsHoweverdisease relevant interactions between these major epigenetic signaling pathways are difficult to study using currently available research toolsHereEpiCypher will develop an innovative recombinant nucleosome based assay platform that leverages interactions between histone and DNA methylation on a single assay substrateThe innovative platform technology developed herein will enable the discovery of novel drug targets as well as prove useful in the identification of context specific inhibitors that are safer and more efficacious than current therapeutics