A common sugar-nucleotide-mediated mechanism of inhibition of (glycosamino)glycan biosynthesis, as evidenced by 6F-GalNAc (Ac3)

FASEB J. 2015 Jul;29(7):2993-3002. doi: 10.1096/fj.14-264226. Epub 2015 Apr 13.

Abstract

Glycosaminoglycan (GAG) polysaccharides have been implicated in a variety of cellular processes, and alterations in their amount and structure have been associated with diseases such as cancer. In this study, we probed 11 sugar analogs for their capacity to interfere with GAG biosynthesis. One analog, with a modification not directly involved in the glycosidic bond formation, 6F-N-acetyl-d-galactosamine (GalNAc) (Ac3), was selected for further study on its metabolic and biologic effect. Treatment of human ovarian carcinoma cells with 50 μM 6F-GalNAc (Ac3) inhibited biosynthesis of GAGs (chondroitin/dermatan sulfate by ∼50-60%, heparan sulfate by ∼35%), N-acetyl-d-glucosamine (GlcNAc)/GalNAc containing glycans recognized by the lectins Datura stramonium and peanut agglutinin (by ∼74 and ∼43%, respectively), and O-GlcNAc protein modification. With respect to function, 6F-GalNAc (Ac3) treatment inhibited growth factor signaling and reduced in vivo angiogenesis by ∼33%. Although the analog was readily transformed in cells into the uridine 5'-diphosphate (UDP)-activated form, it was not incorporated into GAGs. Rather, it strongly reduced cellular UDP-GalNAc and UDP-GlcNAc pools. Together with data from the literature, these findings indicate that nucleotide sugar depletion without incorporation is a common mechanism of sugar analogs for inhibiting GAG/glycan biosynthesis.

Keywords: angiogenesis; glycobiology; growth factor signaling; sugar analog.

Publication types

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

MeSH terms

  • Acetylgalactosamine / analogs & derivatives*
  • Acetylgalactosamine / chemistry
  • Acetylgalactosamine / pharmacology
  • Animals
  • Cell Line
  • Chick Embryo
  • Fibroblast Growth Factor 2 / metabolism
  • Glycosaminoglycans / antagonists & inhibitors
  • Glycosaminoglycans / biosynthesis*
  • HeLa Cells
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Neovascularization, Physiologic / drug effects
  • Polysaccharides / antagonists & inhibitors
  • Polysaccharides / biosynthesis
  • Signal Transduction / drug effects
  • Structure-Activity Relationship
  • Uridine Diphosphate N-Acetylgalactosamine / metabolism
  • Uridine Diphosphate N-Acetylglucosamine / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Glycosaminoglycans
  • Polysaccharides
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Fibroblast Growth Factor 2
  • Uridine Diphosphate N-Acetylglucosamine
  • Uridine Diphosphate N-Acetylgalactosamine
  • Acetylgalactosamine