pevonedistat and Inflammatory-Bowel-Diseases

Neddylation, a newly identified post-translational modification, is significant for the activity and stability of target proteins. The exact role of neddylation in the pathogenesis of inflammatory bowel disease, specifically those mediated by dendritic cells (DCs), was still rarely reported. Here, we showed that inhibition of neddylation protected mice from mucosal inflammation. Targeting neddylation also inhibited DC maturation characterized by reduced cytokine production, down-regulated costimulatory molecules and suppressed capacity in allogeneic T cell stimulation. Additionally, inactivation of neddylation promotes caspase dependent apoptosis of DCs. These phenomena were attributed to the inactivation of mTOR, which was caused by Cullin-1 deneddylation induced Deptor accumulation. Together, our findings revealed that neddylation inhibition suppressed DC functions through mTOR signaling pathway and provided a potential therapeutic opportunity in inflammatory bowel diseases.

Topics: Animals; Apoptosis; Caspase 3; Caspase 7; Cell Differentiation; Cells, Cultured; Cullin Proteins; Cyclopentanes; Cytokines; Dendritic Cells; Disease Models, Animal; Down-Regulation; Enzyme Inhibitors; Humans; Inflammatory Bowel Diseases; Intracellular Signaling Peptides and Proteins; Male; Mice; Mice, Inbred C57BL; Protein Processing, Post-Translational; Pyrimidines; Signal Transduction; TOR Serine-Threonine Kinases

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.

Topics: Animals; Cell Line; Cullin Proteins; Cyclopentanes; Disease Models, Animal; Endopeptidases; Gene Knockdown Techniques; HeLa Cells; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Inflammatory Bowel Diseases; Intestinal Mucosa; Metabolic Networks and Pathways; Mice, Inbred C57BL; NEDD8 Protein; Protease Inhibitors; Protein Stability; Pyrimidines; Ubiquitins