Dissecting neural differentiation regulatory networks through epigenetic footprinting

Nature. 2015 Feb 19;518(7539):355-359. doi: 10.1038/nature13990. Epub 2014 Dec 24.

Abstract

Models derived from human pluripotent stem cells that accurately recapitulate neural development in vitro and allow for the generation of specific neuronal subtypes are of major interest to the stem cell and biomedical community. Notch signalling, particularly through the Notch effector HES5, is a major pathway critical for the onset and maintenance of neural progenitor cells in the embryonic and adult nervous system. Here we report the transcriptional and epigenomic analysis of six consecutive neural progenitor cell stages derived from a HES5::eGFP reporter human embryonic stem cell line. Using this system, we aimed to model cell-fate decisions including specification, expansion and patterning during the ontogeny of cortical neural stem and progenitor cells. In order to dissect regulatory mechanisms that orchestrate the stage-specific differentiation process, we developed a computational framework to infer key regulators of each cell-state transition based on the progressive remodelling of the epigenetic landscape and then validated these through a pooled short hairpin RNA screen. We were also able to refine our previous observations on epigenetic priming at transcription factor binding sites and suggest here that they are mediated by combinations of core and stage-specific factors. Taken together, we demonstrate the utility of our system and outline a general framework, not limited to the context of the neural lineage, to dissect regulatory circuits of differentiation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Cell Differentiation / genetics*
  • Cell Lineage / genetics
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Epigenesis, Genetic / genetics*
  • Epigenomics / methods*
  • Humans
  • Neural Stem Cells / cytology*
  • Neural Stem Cells / metabolism*
  • RNA, Small Interfering / analysis
  • RNA, Small Interfering / genetics
  • Reproducibility of Results
  • Transcription Factors / metabolism
  • Transcription, Genetic / genetics

Substances

  • RNA, Small Interfering
  • Transcription Factors

Associated data

  • GEO/GSE62193