Differentiation of human telencephalic progenitor cells into MSNs by inducible expression of Gsx2 and Ebf1

Proc Natl Acad Sci U S A. 2017 Feb 14;114(7):E1234-E1242. doi: 10.1073/pnas.1611473114. Epub 2017 Jan 30.

Abstract

Medium spiny neurons (MSNs) are a key population in the basal ganglia network, and their degeneration causes a severe neurodegenerative disorder, Huntington's disease. Understanding how ventral neuroepithelial progenitors differentiate into MSNs is critical for regenerative medicine to develop specific differentiation protocols using human pluripotent stem cells. Studies performed in murine models have identified some transcriptional determinants, including GS Homeobox 2 (Gsx2) and Early B-cell factor 1 (Ebf1). Here, we have generated human embryonic stem (hES) cell lines inducible for these transcription factors, with the aims of (i) studying their biological role in human neural progenitors and (ii) incorporating TF conditional expression in a developmental-based protocol for generating MSNs from hES cells. Using this approach, we found that Gsx2 delays cell-cycle exit and reduces Pax6 expression, whereas Ebf1 promotes neuronal differentiation. Moreover, we found that Gsx2 and Ebf1 combined overexpression in hES cells achieves high yields of MSNs, expressing Darpp32 and Ctip2, in vitro as well in vivo after transplantation. We show that hES-derived striatal progenitors can be transplanted in animal models and can differentiate and integrate into the host, extending fibers over a long distance.

Keywords: Ebf1; Gsx2; HD; MSNs; hES cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Cycle / genetics
  • Cell Differentiation / genetics*
  • Cell Line
  • Dopamine and cAMP-Regulated Phosphoprotein 32 / genetics
  • Dopamine and cAMP-Regulated Phosphoprotein 32 / metabolism
  • Gene Expression
  • Homeodomain Proteins / genetics*
  • Homeodomain Proteins / metabolism
  • Human Embryonic Stem Cells / metabolism*
  • Human Embryonic Stem Cells / transplantation
  • Humans
  • Mice, Nude
  • Neurons / cytology
  • Neurons / metabolism*
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Stem Cell Transplantation / methods
  • Telencephalon / cytology
  • Trans-Activators / genetics*
  • Trans-Activators / metabolism
  • Transplantation, Heterologous
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / metabolism

Substances

  • BCL11B protein, human
  • Dopamine and cAMP-Regulated Phosphoprotein 32
  • EBF1 protein, human
  • GSH2 protein, human
  • Homeodomain Proteins
  • Repressor Proteins
  • Trans-Activators
  • Tumor Suppressor Proteins