Stabilization of HIF through inhibition of Cullin-2 neddylation is protective in mucosal inflammatory responses

FASEB J. 2015 Jan;29(1):208-15. doi: 10.1096/fj.14-259663. Epub 2014 Oct 17.


There is interest in understanding post-translational modifications of proteins in inflammatory disease. Neddylation is the conjugation of the molecule neural precursor cell expressed, developmentally down-regulated 8 (NEDD8) to promote protein stabilization. Cullins are a family of NEDD8 targets important in the stabilization and degradation of proteins, such as hypoxia-inducible factor (HIF; via Cullin-2). Here, we elucidate the role of human deneddylase-1 (DEN-1, also called SENP8) in inflammatory responses in vitro and in vivo and define conditions for targeting neddylation in models of mucosal inflammation. HIF provides protection in inflammatory models, so we examined the contribution of DEN-1 to HIF stabilization. Pharmacologic targeting of neddylation activity with MLN4924 (IC50, 4.7 nM) stabilized HIF-1α, activated HIF promoter activity by 2.5-fold, and induced HIF-target genes in human epithelial cells up to 5-fold. Knockdown of DEN-1 in human intestinal epithelial cells resulted in increased kinetics in barrier formation, decreased permeability, and enhanced barrier restitution by 2 ± 0.5-fold. Parallel studies in vivo revealed that MLN4924 abrogated disease severity in murine dextran sulfate sodium colitis, including weight loss, colon length, and histologic severity. We conclude that DEN-1 is a regulator of cullin neddylation and fine-tunes the inflammatory response in vitro and in vivo. Pharmacologic inhibition of cullin neddylation may provide a therapeutic opportunity in mucosal inflammatory disease.

Keywords: hypoxia; inflammation; intestinal epithelia.

Publication types

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

MeSH terms

  • Animals
  • Cell Line
  • Cullin Proteins / antagonists & inhibitors
  • Cullin Proteins / metabolism*
  • Cyclopentanes / pharmacology
  • Disease Models, Animal
  • Endopeptidases / genetics
  • Endopeptidases / metabolism
  • Gene Knockdown Techniques
  • HeLa Cells
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • Inflammatory Bowel Diseases / metabolism*
  • Inflammatory Bowel Diseases / pathology
  • Inflammatory Bowel Diseases / prevention & control*
  • Intestinal Mucosa / metabolism
  • Intestinal Mucosa / pathology
  • Metabolic Networks and Pathways
  • Mice, Inbred C57BL
  • NEDD8 Protein
  • Protease Inhibitors / pharmacology
  • Protein Stability
  • Pyrimidines / pharmacology
  • Ubiquitins / metabolism


  • CUL2 protein, human
  • Cullin Proteins
  • Cyclopentanes
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • NEDD8 Protein
  • NEDD8 protein, human
  • Protease Inhibitors
  • Pyrimidines
  • Ubiquitins
  • Endopeptidases
  • SENP8 protein, human
  • pevonedistat