High throughput screening for inhibitors of REST in neural derivatives of human embryonic stem cells reveals a chemical compound that promotes expression of neuronal genes

Stem Cells. 2013 Sep;31(9):1816-28. doi: 10.1002/stem.1430.

Abstract

Decreased expression of neuronal genes such as brain-derived neurotrophic factor (BDNF) is associated with several neurological disorders. One molecular mechanism associated with Huntington disease (HD) is a discrete increase in the nuclear activity of the transcriptional repressor REST/NRSF binding to repressor element-1 (RE1) sequences. High-throughput screening of a library of 6,984 compounds with luciferase-assay measuring REST activity in neural derivatives of human embryonic stem cells led to identify two benzoimidazole-5-carboxamide derivatives that inhibited REST silencing in a RE1-dependent manner. The most potent compound, X5050, targeted REST degradation, but neither REST expression, RNA splicing nor binding to RE1 sequence. Differential transcriptomic analysis revealed the upregulation of neuronal genes targeted by REST in wild-type neural cells treated with X5050. This activity was confirmed in neural cells produced from human induced pluripotent stem cells derived from a HD patient. Acute intraventricular delivery of X5050 increased the expressions of BDNF and several other REST-regulated genes in the prefrontal cortex of mice with quinolinate-induced striatal lesions. This study demonstrates that the use of pluripotent stem cell derivatives can represent a crucial step toward the identification of pharmacological compounds with therapeutic potential in neurological affections involving decreased expression of neuronal genes associated to increased REST activity, such as Huntington disease.

Keywords: High throughput screening; Human embryonic stem cells; Neural stem cells; Neuron restrictive silencer factor; Repressor element-1 silencing transcription factor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Disease Models, Animal
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects
  • Embryonic Stem Cells / metabolism*
  • Gene Expression Regulation / drug effects*
  • Genes, Reporter
  • High-Throughput Screening Assays / methods*
  • Humans
  • Huntington Disease / pathology
  • Luciferases / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / metabolism*
  • Neurons / drug effects
  • Neurons / metabolism*
  • Repressor Proteins / antagonists & inhibitors*
  • Repressor Proteins / metabolism
  • Small Molecule Libraries / pharmacology*
  • Transcriptome / drug effects
  • Transcriptome / genetics

Substances

  • RE1-silencing transcription factor
  • Repressor Proteins
  • Small Molecule Libraries
  • Luciferases