RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin

ABSTRACT Clostridium perfringens type F strains cause gastrointestinal disease when they produce a pore-forming toxin named C. perfringens enterotoxin (CPE). In human enterocyte-like Caco-2 cells, low CPE concentrations cause caspase-3-dependent apoptosis, while high CPE concentrations cause necrosi...

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Autores principales:	Archana Shrestha, Iman Mehdizadeh Gohari, Bruce A. McClane
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Clostridium perfringens enterotoxin apoptosis necroptosis RIP1 kinase RIP3 kinase MLKL Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/e5435ed5905e4157a311ebfe2783d3df
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spelling	oai:doaj.org-article:e5435ed5905e4157a311ebfe2783d3df2021-11-15T15:54:44ZRIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin10.1128/mBio.02985-192150-7511https://doaj.org/article/e5435ed5905e4157a311ebfe2783d3df2019-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02985-19https://doaj.org/toc/2150-7511ABSTRACT Clostridium perfringens type F strains cause gastrointestinal disease when they produce a pore-forming toxin named C. perfringens enterotoxin (CPE). In human enterocyte-like Caco-2 cells, low CPE concentrations cause caspase-3-dependent apoptosis, while high CPE concentrations cause necrosis. Since necrosis or apoptosis sometimes involves receptor-interacting serine/threonine-protein kinase-1 or 3 (RIP1 or RIP3), this study examined whether those kinases are important for CPE-induced apoptosis or necrosis. Highly specific RIP1 or RIP3 inhibitors reduced both CPE-induced apoptosis and necrosis in Caco-2 cells. Those findings suggested that the form of necrosis induced by treating Caco-2 cells with high CPE concentrations involves necroptosis, which was confirmed when high, but not low, CPE concentrations were shown to induce oligomerization of mixed-lineage kinase domain-like pseudokinase (MLKL), a key late step in necroptosis. Furthermore, an MLKL oligomerization inhibitor reduced cell death caused by high, but not low, CPE concentrations. Supporting RIP1 and RIP3 involvement in CPE-induced necroptosis, inhibitors of those kinases also reduced MLKL oligomerization during treatment with high CPE concentrations. Calpain inhibitors similarly blocked MLKL oligomerization induced by high CPE concentrations, implicating calpain activation as a key intermediate in initiating CPE-induced necroptosis. In two other CPE-sensitive cell lines, i.e., Vero cells and human enterocyte-like T84 cells, low CPE concentrations also caused primarily apoptosis/late apoptosis, while high CPE concentrations mainly induced necroptosis. Collectively, these results establish that high, but not low, CPE concentrations cause necroptosis and suggest that RIP1, RIP3, MLKL, or calpain inhibitors can be explored as potential therapeutics against CPE effects in vivo. IMPORTANCE C. perfringens type F strains are a common cause of food poisoning and antibiotic-associated diarrhea. Type F strain virulence requires production of C. perfringens enterotoxin (CPE). In Caco-2 cells, high CPE concentrations cause necrosis while low enterotoxin concentrations induce apoptosis. The current study determined that receptor-interacting serine/threonine-protein kinases 1 and 3 are involved in both CPE-induced apoptosis and necrosis in Caco-2 cells, while mixed-lineage kinase domain-like pseudokinase (MLKL) oligomerization is involved in CPE-induced necrosis, thereby indicating that this form of CPE-induced cell death involves necroptosis. High CPE concentrations also caused necroptosis in T84 and Vero cells. Calpain activation was identified as a key intermediate for CPE-induced necroptosis. These results suggest inhibitors of RIP1, RIP3, MLKL oligomerization, or calpain are useful therapeutics against CPE-mediated diseases.Archana ShresthaIman Mehdizadeh GohariBruce A. McClaneAmerican Society for MicrobiologyarticleClostridium perfringens enterotoxinapoptosisnecroptosisRIP1 kinaseRIP3 kinaseMLKLMicrobiologyQR1-502ENmBio, Vol 10, Iss 6 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Clostridium perfringens enterotoxin apoptosis necroptosis RIP1 kinase RIP3 kinase MLKL Microbiology QR1-502
spellingShingle	Clostridium perfringens enterotoxin apoptosis necroptosis RIP1 kinase RIP3 kinase MLKL Microbiology QR1-502 Archana Shrestha Iman Mehdizadeh Gohari Bruce A. McClane RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin
description	ABSTRACT Clostridium perfringens type F strains cause gastrointestinal disease when they produce a pore-forming toxin named C. perfringens enterotoxin (CPE). In human enterocyte-like Caco-2 cells, low CPE concentrations cause caspase-3-dependent apoptosis, while high CPE concentrations cause necrosis. Since necrosis or apoptosis sometimes involves receptor-interacting serine/threonine-protein kinase-1 or 3 (RIP1 or RIP3), this study examined whether those kinases are important for CPE-induced apoptosis or necrosis. Highly specific RIP1 or RIP3 inhibitors reduced both CPE-induced apoptosis and necrosis in Caco-2 cells. Those findings suggested that the form of necrosis induced by treating Caco-2 cells with high CPE concentrations involves necroptosis, which was confirmed when high, but not low, CPE concentrations were shown to induce oligomerization of mixed-lineage kinase domain-like pseudokinase (MLKL), a key late step in necroptosis. Furthermore, an MLKL oligomerization inhibitor reduced cell death caused by high, but not low, CPE concentrations. Supporting RIP1 and RIP3 involvement in CPE-induced necroptosis, inhibitors of those kinases also reduced MLKL oligomerization during treatment with high CPE concentrations. Calpain inhibitors similarly blocked MLKL oligomerization induced by high CPE concentrations, implicating calpain activation as a key intermediate in initiating CPE-induced necroptosis. In two other CPE-sensitive cell lines, i.e., Vero cells and human enterocyte-like T84 cells, low CPE concentrations also caused primarily apoptosis/late apoptosis, while high CPE concentrations mainly induced necroptosis. Collectively, these results establish that high, but not low, CPE concentrations cause necroptosis and suggest that RIP1, RIP3, MLKL, or calpain inhibitors can be explored as potential therapeutics against CPE effects in vivo. IMPORTANCE C. perfringens type F strains are a common cause of food poisoning and antibiotic-associated diarrhea. Type F strain virulence requires production of C. perfringens enterotoxin (CPE). In Caco-2 cells, high CPE concentrations cause necrosis while low enterotoxin concentrations induce apoptosis. The current study determined that receptor-interacting serine/threonine-protein kinases 1 and 3 are involved in both CPE-induced apoptosis and necrosis in Caco-2 cells, while mixed-lineage kinase domain-like pseudokinase (MLKL) oligomerization is involved in CPE-induced necrosis, thereby indicating that this form of CPE-induced cell death involves necroptosis. High CPE concentrations also caused necroptosis in T84 and Vero cells. Calpain activation was identified as a key intermediate for CPE-induced necroptosis. These results suggest inhibitors of RIP1, RIP3, MLKL oligomerization, or calpain are useful therapeutics against CPE-mediated diseases.
format	article
author	Archana Shrestha Iman Mehdizadeh Gohari Bruce A. McClane
author_facet	Archana Shrestha Iman Mehdizadeh Gohari Bruce A. McClane
author_sort	Archana Shrestha
title	RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin
title_short	RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin
title_full	RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin
title_fullStr	RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin
title_full_unstemmed	RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin
title_sort	rip1, rip3, and mlkl contribute to cell death caused by <named-content content-type="genus-species">clostridium perfringens</named-content> enterotoxin
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/e5435ed5905e4157a311ebfe2783d3df
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RIP1, RIP3, and MLKL Contribute to Cell Death Caused by <named-content content-type="genus-species">Clostridium perfringens</named-content> Enterotoxin

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