Abstract B015: Versatile biochemical and genomic platforms for drug discovery in chromatin remodeling research


 Chromatin remodeling, driven by ATP-dependent remodeling enzymes, plays key roles in regulating gene expression, genome replication and repair. Aberrant nucleosome organization can severely disrupt these important processes, driving various cancers and developmental disorders. Remarkably, nearly 20% of all human cancers contain mutations in subunits from the SWI/SNF family of chromatin remodeling complexes, making them attractive therapeutic targets and interests of basic research. However, studies on the remodeling enzymes (and their multi-subunit complexes) in vitro are challenging partially due to the requirement for nucleosome-based substrates. We have created the EpiDyne® nucleosome remodeling substrate portfolio for use in biochemical assays to examine nucleosome immobilizing activities of chromatin remodelers. Here we present the development of multiple remodeling assays on the EpiDyne® platform (EpiDyne®-FRET, EpiDyne®-PicoGreenTM and EpiDyne®-TR-FRET). These nonradioactive plate-based assays are easy to use, ready for high-throughput inhibitor screening, and can be customized for various remodeling enzymes that exhibit distinct preferences in nucleosome compositions. Furthermore, genome-wide mapping of chromatin remodelers is fundamental for understanding their functions in diseases and responses to inhibitors in cells, but is often obscured by unreliable ChIP-seq approaches. To this end, we have optimized the CUTANA™ CUT&RUN approach to efficiently capture localizations of all major classes of chromatin remodelers with high signal to background. As a complementary research tool to the EpiDyne® platform, CUT&RUN facilitates the epigenomic research on chromatin remodelers for drug discoveries.
 Citation Format: Natalia L. Husby, Lu Sun, Hannah Willis, Matthew R. Marunde, Matt J. Meiners, Zachary Gillespie, Rachel Watson, Saarang Gopinath, Jonathan M. Burg, Marcus A. Cheek, Katherine Novitzky, Bryan J. Venters, Zu-Wen Sun, Michael-Christopher Keogh. Versatile biochemical and genomic platforms for drug discovery in chromatin remodeling research. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Epigenomics; 2022 Oct 6-8; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_2):Abstract nr B015.