Perturbation of neddylation-dependent NF-κB responses in the intestinal epithelium drives apoptosis and inhibits resolution of mucosal inflammation.
Recent work has revealed a central role for neddylation (the conjugation of a Nedd8-moiety to Cullin proteins) in the fine tuning of the NF-κB response (via Cullin-1). In the present study, we investigated the contribution of Cullin-1 neddylation and NF-κB signaling to mucosal inflammatory responses in vitro and in vivo. Initial in vitro studies using cultured intestinal epithelial cells revealed that the neddylation inhibitor MLN4924 prominently induces the deneddylation of Cullin-1. Parallel western blot, luciferase reporter and gene target assays identified MLN4924 as a potent inhibitor of intestinal epithelial NF-κB. Subsequent studies revealed that MLN4924 potently induces epithelial apoptosis but only in the presence of additional inflammatory stimuli. In vivo administration of MLN4924 (3 mg/kg/d) in a TNBS-induce colitis model significantly accentuated disease severity. Indeed, MLN4924 resulted in worsened clinical scores and increased mortality early in the inflammatory response. Histologic analysis of the colon revealed that neddylation inhibition results in increased tissue damage and significantly increased mucosal apoptosis as determined by TUNEL and cleaved caspase-3 staining, particularly prominent within the epithelium. Extensions of these studies revealed that ongoing inflammation is associated with significant loss of deneddylase-1 (SENP8) expresssion. These studies reveal that intact Cullin-1 neddylation is central to resolution of acute inflammation.
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045 Department of Anesthesiology, University Hospital Bonn, Germany.
Stefan F
SF
Ehrentraut
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045.
Valerie F
VF
Curtis
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045.
Ruth X
RX
Wang
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045.
Bejan J
BJ
Saeedi
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045 Department of Anesthesiology, University Hospital Bonn, Germany.
Heidi
H
Ehrentraut
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045 Eastern Colorado Veterans Affairs Hospital, Denver, CO.
Joseph C
JC
Onyiah
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045.
Caleb J
CJ
Kelly
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045.
Eric L
EL
Campbell
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045.
Louise E
LE
Glover
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045 Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717.
Douglas J
DJ
Kominsky
Mucosal Inflammation Program, Department of Medicine and Immunology, University of Colorado School of Medicine, Aurora, CO 80045 Eastern Colorado Veterans Affairs Hospital, Denver, CO Sean.Colgan@UCDenver.edu.
Sean P
SP
Colgan
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