Engineered Glycocalyx Regulates Stem Cell Proliferation in Murine Crypt Organoids

Cell Chem Biol. 2018 Apr 19;25(4):439-446.e5. doi: 10.1016/j.chembiol.2018.01.010. Epub 2018 Feb 8.

Abstract

At the base of the intestinal crypt, long-lived Lgr5+ stem cells are intercalated by Paneth cells that provide essential niche signals for stem cell maintenance. This unique epithelial anatomy makes the intestinal crypt one of the most accessible models for the study of adult stem cell biology. The glycosylation patterns of this compartment are poorly characterized, and the impact of glycans on stem cell differentiation remains largely unexplored. We find that Paneth cells, but not Lgr5+ stem cells, express abundant terminal N-acetyllactosamine (LacNAc). Employing an enzymatic method to edit glycans in cultured crypt organoids, we assess the functional role of LacNAc in the intestinal crypt. We discover that blocking access to LacNAc on Paneth cells leads to hyperproliferation of the neighboring Lgr5+ stem cells, which is accompanied by the downregulation of genes that are known as negative regulators of proliferation.

Keywords: LacNAc; Paneth cells; enteroid; enzymatic glycan engineering; glycosylation; intestinal stem cell.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Sugars / analysis
  • Amino Sugars / metabolism*
  • Animals
  • Cell Differentiation
  • Cell Proliferation*
  • Cells, Cultured
  • Glycocalyx / metabolism*
  • Humans
  • Mice, Inbred C57BL
  • Organoids / cytology*
  • Organoids / metabolism
  • Paneth Cells / cytology*
  • Paneth Cells / metabolism
  • Receptors, G-Protein-Coupled / analysis
  • Stem Cells / cytology*
  • Stem Cells / metabolism

Substances

  • Amino Sugars
  • Lgr5 protein, mouse
  • Receptors, G-Protein-Coupled
  • N-acetyllactosamine