title: Identification of HDAC Inhibitors Using a Cell-Based HDAC I/II Assay.
abstract: Histone deacetylases (HDACs) are a class of epigenetic enzymes that regulate gene expression by histone deacetylation. Altered HDAC function has been linked to cancer and neurodegenerative diseases, making HDACs popular therapeutic targets. In this study, we describe a screening approach for identification of compounds that inhibit endogenous class I and II HDACs. A homogeneous, luminogenic HDAC I/II assay was optimized in a 1536-well plate format in several human cancer cell lines, including HCT116 and human neural stem cells. The assay confirmed 37 known HDAC inhibitors from two libraries of known epigenetics-active compounds. Using the assay, we identified a group of potential HDAC inhibitors by screening the National Center for Advancing Translational Sciences (NCATS) Pharmaceutical Collection of 2527 small-molecule drugs. The selected compounds showed similar HDAC I/II inhibitory potency and efficacy values in both HCT116 and neural stem cells. Several previously unidentified HDAC inhibitors were further evaluated and profiled for their selectivity against a panel of 10 HDAC I/II isoforms using fluorogenic HDAC biochemical assays. In summary, our results show that several novel HDAC inhibitors, including nafamostat and piceatannol, have been identified using the HDAC I/II cell-based assay, and multiple cell types have been validated for high-throughput screening of large chemical libraries.
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA.
authors: Chia-Wen
authors: CW
authors: Hsu
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA.
authors: David
authors: D
authors: Shou
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA.
authors: Ruili
authors: R
authors: Huang
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA.
authors: Thai
authors: T
authors: Khuc
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA.
authors: Sheng
authors: S
authors: Dai
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA.
authors: Wei
authors: W
authors: Zheng
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA.
authors: Carleen
authors: C
authors: Klumpp-Thomas
authors: National Center for Advancing Translational Sciences, Bethesda, MD, USA mxia@mail.nih.gov.
authors: Menghang
authors: M
authors: Xia
mesh_terms: D000107:Acetylation / Q000187:drug effects; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D045325:HCT116 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D056572:Histone Deacetylase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D058953:Neural Stem Cells / Q000187:drug effects
chemical_list: D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases
keywords: cancer; epigenetics; histone deacetylase; neurodegenerative disease; qHTS