Anna Negroni1, Eleonora Colantoni2, Maria Pierdomenico2, Francesca Palone2, Manuela Costanzo2, Salvatore Oliva2, Antonio Tiberti3, Salvatore Cucchiara2, Laura Stronati4. 1. Division of Health Protection Technologies, Territorial and Production Systems Sustainability Department, ENEA, Rome, Italy. Electronic address: anna.negroni@enea.it. 2. Department of Pediatrics and Infantile Neuropsychiatry, Pediatric Gastroenterology and Liver Unit, Sapienza University of Rome, Italy. 3. Department of Internal Medicine and Medical Specialties, Sapienza University of Rome, Rome, Italy. 4. Department of Cellular Biotechnology and Hematology, Sapienza University of Rome, Rome, Italy.
Abstract
BACKGROUND: Necroptosis is an inflammatory form of programmed cell death requiring receptor-interacting protein kinase 3 (RIP3) and mixed lineage kinase domain-like protein (MLKL). AIMS: The aim of this study is to examine in depth in vitro and ex vivo the contribution of necroptosis to intestinal inflammation. METHODS: In vitro: we used an intestinal cell line, HCT116RIP3, produced in our laboratory and overexpressing RIP3. Ex vivo: intestinal mucosal biopsies were taken from patients with inflammatory bowel disease (IBD) (20 with Crohn's disease; 20 with ulcerative colitis) and from 20 controls. RESULTS: RIP3-induced necroptosis triggers MLKL activation, increases cytokine/alarmin expression (IL-8, IL-1β, IL-33, HMGB1), NF-kBp65 translocation and NALP3 inflammasome assembly. It also affects membrane permeability by altering cell-cell junctional proteins (E-cadherin, Occludin, Zonulin-1). Targeting necroptosis through Necrostatin-1 significantly reduces intestinal inflammation in vitro and in cultured intestinal explants from IBD. CONCLUSION: We show for the first time in vitro and ex vivo that RIP3-driven necroptosis seriously affects intestinal inflammation by increasing pMLKL, activating different cytokines and alarmins, and altering epithelial permeability. The inhibition of necroptosis causes a significant decrease of all these effects. These data strongly support the view that targeting necroptosis may represent a promising new option for the treatment of inflammatory enteropathies.
BACKGROUND: Necroptosis is an inflammatory form of programmed cell death requiring receptor-interacting protein kinase 3 (RIP3) and mixed lineage kinase domain-like protein (MLKL). AIMS: The aim of this study is to examine in depth in vitro and ex vivo the contribution of necroptosis to intestinal inflammation. METHODS: In vitro: we used an intestinal cell line, HCT116RIP3, produced in our laboratory and overexpressing RIP3. Ex vivo: intestinal mucosal biopsies were taken from patients with inflammatory bowel disease (IBD) (20 with Crohn's disease; 20 with ulcerative colitis) and from 20 controls. RESULTS:RIP3-induced necroptosis triggers MLKL activation, increases cytokine/alarmin expression (IL-8, IL-1β, IL-33, HMGB1), NF-kBp65 translocation and NALP3 inflammasome assembly. It also affects membrane permeability by altering cell-cell junctional proteins (E-cadherin, Occludin, Zonulin-1). Targeting necroptosis through Necrostatin-1 significantly reduces intestinal inflammation in vitro and in cultured intestinal explants from IBD. CONCLUSION: We show for the first time in vitro and ex vivo that RIP3-driven necroptosis seriously affects intestinal inflammation by increasing pMLKL, activating different cytokines and alarmins, and altering epithelial permeability. The inhibition of necroptosis causes a significant decrease of all these effects. These data strongly support the view that targeting necroptosis may represent a promising new option for the treatment of inflammatory enteropathies.
Authors: Stefanie M Bader; Simon P Preston; Katie Saliba; Adam Lipszyc; Zoe L Grant; Liana Mackiewicz; Andrew Baldi; Anne Hempel; Michelle P Clark; Thanushi Peiris; William Clow; Jan Bjelic; Michael D Stutz; Philip Arandjelovic; Jack Teale; Fashuo Du; Leigh Coultas; James M Murphy; Cody C Allison; Marc Pellegrini; Andre L Samson Journal: Cell Death Differ Date: 2022-07-23 Impact factor: 12.067
Authors: Danielle Pretorius; Asher M Kahn-Krell; Wesley C LaBarge; Xi Lou; Ramaswamy Kannappan; Andrew E Pollard; Vladimir G Fast; Joel L Berry; Alan W Eberhardt; Jianyi Zhang Journal: Biomed Mater Date: 2021-02-26 Impact factor: 3.715
Authors: Eva Alegre-Cortés; Alicia Muriel-González; Saray Canales-Cortés; Elisabet Uribe-Carretero; Guadalupe Martínez-Chacón; Ana Aiastui; Adolfo López de Munain; Mireia Niso-Santano; Rosa A Gonzalez-Polo; José M Fuentes; Sokhna M S Yakhine-Diop Journal: Antioxidants (Basel) Date: 2020-06-15
Authors: Asher Kahn-Krell; Danielle Pretorius; Jianfa Ou; Vladimir G Fast; Silvio Litovsky; Joel Berry; Xiaoguang Margaret Liu; Jianyi Zhang Journal: Front Bioeng Biotechnol Date: 2021-06-11