RIPK1 Mediates TNF-Induced Intestinal Crypt Apoptosis During Chronic NF-κB Activation
Wong, Jerry (Department Pathology, San Diego, California)
Garcia-Carbonell, Ricard (Universitat de Barcelona. Departament de Bioquímica i Biomedicina Molecular)
Zelic, Matija (University of Massachusetts Medical School)
Ho, Samuel B. (VA San Diego Healthcare System, San Diego, California)
Boland, Brigid S. (San Diego Digestive Disease Research Center, San Diego, California)
Yao, Shih-Jing (Department Pathology, San Diego, California)
Desai, Shalin A. (Department Pathology, San Diego, California)
Das, Soumita (Department Pathology, San Diego, California)
Planell, Núria (Institut d'Investigacions Biomèdiques August Pi i Sunyer)
Harris, Philip A. (Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania)
Font-Burgada, Joan (Department Pathology, San Diego, California)
Taniguchi, Koji (Department Pathology, San Diego, California)
Bertin, John (University of Massachusetts Medical School)
Salas, Azucena (Institut d'Investigacions Biomèdiques August Pi i Sunyer)
Pasparakis, Manolis (University of Cologne, Cologne, Germany)
Gough, Pete J. (Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania)
Kelliher, Michelle (University of Massachusetts Medical School)
Karin, Michael (Department Pathology, San Diego, California)
Guma, Monica 1971- (Universitat Autònoma de Barcelona. Departament de Medicina)

Date:	2019
Abstract:	Tumor necrosis factor (TNF) is a major pathogenic effector and a therapeutic target in inflammatory bowel disease (IBD), yet the basis for TNF-induced intestinal epithelial cell (IEC) death is unknown, because TNF does not kill normal IECs. Here, we investigated how chronic nuclear factor (NF)- κB activation, which occurs in human IBD, promotes TNF-dependent IEC death in mice. Human IBD specimens were stained for p65 and cleaved caspase-3. C57BL/6 mice with constitutively active IKKβ in IEC (Ikkβ(EE) IEC ), Ripk1 D138N/D138N knockin mice, and Ripk3 -/- mice were injected with TNF or lipopolysaccharide. Enteroids were also isolated from these mice and challenged with TNF with or without RIPK1 and RIPK3 inhibitors or butylated hydroxyanisole. Ripoptosome-mediated caspase-8 activation was assessed by immunoprecipitation. NF-κB activation in human IBD correlated with appearance of cleaved caspase-3. Congruently, unlike normal mouse IECs that are TNF-resistant, IECs in Ikkβ(EE) IEC mice and enteroids were susceptible to TNF-dependent apoptosis, which depended on the protein kinase function of RIPK1. Constitutively active IKKβ facilitated ripoptosome formation, a RIPK1 signaling complex that mediates caspase-8 activation by TNF. Butylated hydroxyanisole treatment and RIPK1 inhibitors attenuated TNF-induced and ripoptosome-mediated caspase-8 activation and IEC death in vitro and in vivo. Contrary to common expectations, chronic NF-κB activation induced intestinal crypt apoptosis after TNF stimulation, resulting in severe mucosal erosion. RIPK1 kinase inhibitors selectively inhibited TNF destructive properties while preserving its survival and proliferative properties, which do not require RIPK1 kinase activity. RIPK1 kinase inhibition could be a potential treatment for IBD.
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	IBD ; RIPK1 ; Intestinal Epithelial Cell ; Ripoptosome ; Cell Death ; BHA, butylated hydroxyanisole ; Cc-3, cleaved caspase-3 ; Cc-8, cleaved caspase-8 ; CD, Crohn's disease ; CHX, cycloheximide ; DPI, phenyleneiodonium ; IB, immunoblotting ; IBD, inflammatory bowel disease ; IEC, intestinal epithelial cell ; IHC, immunohistochemistry ; I.p., intraperitoneally ; LPS, lipopolysaccharide ; Nec-1, necrostatin-1 ; NF, nuclear factor ; PBS, phosphate-buffered saline ; Qpcr, quantitative polymerase chain reaction ; TNF, tumor necrosis factor ; UC, ulcerative colitis ; WT, wild type
Published in:	Cellular and Molecular Gastroenterology and Hepatology, Vol. 9 (october 2019) , p. 295-312, ISSN 2352-345X

DOI: 10.1016/j.jcmgh.2019.10.002
PMID: 31606566

18 p, 7.7 MB

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