MAY 30, 2018 10:30 AM PDT

Inhibitors of NSD2 Discovered by High-Throughput Screening with a Nucleosome Substrate

C.E. CREDITS: P.A.C.E. CE | Florida CE
  • Senior Research Scientist, Biology, Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health
      Dr. Nathan P. Coussens is a Senior Research Scientist in the Division of Pre-Clinical Innovation at the National Center for Advancing Translational Sciences (NCATS). He earned a Ph.D. in biochemistry from the University of Iowa, where he applied structural and biophysical approaches to the study of host-pathogen interactions. As a postdoctoral fellow of the Interdisciplinary Immunology Program at the University of Iowa, he utilized X-ray crystallography and thermodynamic binding studies to inform the development of small molecules that target a host evasion mechanism exploited by families of pathogenic bacteria. Prior to joining NCATS in 2013, Dr. Coussens was a postdoctoral fellow at the National Cancer Institute where he combined biophysical studies, cell biology, and high-resolution imaging to interrogate molecular signaling events initiated by the T cell antigen receptor. At NCATS, Dr. Coussens applies his diverse scientific background to develop and optimize novel biochemical and cell-based assays for high-throughput screening. He works with a highly collaborative and multidisciplinary team to develop small molecule probes relevant to a variety of human diseases. Dr. Coussens also serves as editor-in-chief of the Assay Guidance Manual, a growing online eBook of best practices for preclinical assay development and implementation that has become the go-to resource for scientists in industry and academia.


    NSD2 catalyzes the mono- and di-methylation of the e-amine of lysine 36 from histone H3, utilizing the methyl donor S-adenosyl-L-methionine. Increased catalytic activity of NSD2, either by overexpression or point mutations, is associated with multiple human cancers. To date, potent, selective and cell-active NSD2 inhibitors have not been described. A platform was established for the discovery of selective NSD2 inhibitors from quantitative high-throughput screening in a 1536-well format using a nucleosome substrate. Active compounds from the primary screen were triaged by a panel of assays and tested against two clinically relevant NSD2 mutants, E1099K and T1150A. Five confirmed inhibitors were further evaluated by a radiolabeled validation assay, surface plasmon resonance studies, methyltransferase profiling, and histone methylation in U-2 OS cells. Several inhibitors bound the NSD2 catalytic SET domain and demonstrated activity in cells, which validates the use of this workflow for the discovery and development of selective NSD2 inhibitors.

    Learning Objectives: 

    1. The importance of orthogonal and counter assays for hit selection
    2. The value of assay miniaturization for expensive/limited reagents

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