Chemical Modulation of Protein O-GlcNAcylation via OGT Inhibition Promotes Human Neural Cell Differentiation

  • ACS Chem Biol. 2017 Aug 18;12(8):2030-2039. doi: 10.1021/acschembio.7b00232.
Lissette M Andres  1 ,  Ian W Blong  2 ,  Angela C Evans  3 ,  Neil G Rumachik  4 ,  Teppei Yamaguchi  1 ,  Nam D Pham  5 ,  Pamela Thompson  3 ,  Jennifer J Kohler  5 ,  Carolyn R Bertozzi  4  6
Affiliations
  • 1. Department of Molecular and Cell Biology, University of California , Berkeley, California 94720, United States.
  • 2. Department of Stem Cell Biology and Regenerative Medicine, Stanford University , Stanford, California 94305, United States.
  • 3. Department of Chemistry, University of California , Berkeley, California 94720, United States.
  • 4. Department of Chemistry, Stanford University , Stanford, California 94305, United States.
  • 5. Department of Biochemistry, University of Texas Southwestern Medical Center , Dallas, Texas 75390, United States.
  • 6. Howard Hughes Medical Institute , Chevy Chase, Maryland, United States.
PMID: 28541657 DOI: 10.1021/acschembio.7b00232
Abstract

The Enzymes that determine protein O-GlcNAcylation, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), act on key transcriptional and epigenetic regulators, and both are abundantly expressed in the brain. However, little is known about how alterations in O-GlcNAc cycling affect human embryonic stem cell (hESC) neural differentiation. Here, we studied the effects of perturbing O-GlcNAcylation during neural induction of hESCs using the metabolic inhibitor of OGT, peracetylated 5-thio-N-acetylglucosamine (Ac4-5SGlcNAc). Treatment of hESCs with Ac4-5SGlcNAc during induction limited protein O-GlcNAcylation and also caused a dramatic decrease in global levels of UDP-GlcNAc. Concomitantly, a subpopulation of neural progenitor cells (NPCs) acquired an immature neuronal morphology and expressed early neuronal markers such as β-III tubulin (TUJ1) and microtubule associated protein 2 (MAP2), phenotypes that took longer to manifest in the absence of OGT inhibition. These data suggest that chemical inhibition of OGT and perturbation of protein O-GlcNAcylation accelerate the differentiation of hESCs along the neuronal lineage, thus providing further insight into the dynamic molecular mechanisms involved in neuronal development.

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