NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) modulation by intracellular Cl– concentration

Abstract: The impairment of the cystic fibrosis transmembrane conductance regulator (CFTR) activity induces intracellular chloride (Cl– ) accumulation. The anion Cl– , acting as a second messenger, stimulates the secretion of interleukin-1β (IL-1β), which starts an autocrine positive feedback loop. Here, we show that NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) are indirectly modulated by the intracellular Cl– concentration, showing maximal expression and activity at 75 mM Cl– , in the presence of the ionophores nigericin and tributyltin. The expression of PYD and CARD domain containing (PYCARD/ASC) remained constant from 0 to 125 mM Cl– . The CASP1 inhibitor VX-765 and the NLRP3 inflammasome inhibitor MCC950 completely blocked the Cl– - stimulated IL-1β mRNA expression and partially the IL-1β secretion. DCF fluorescence (cellular reactive oxygen species, cROS) and MitoSOX fluorescence (mitochondrial ROS, mtROS) also showed maximal ROS levels at 75 mM Cl– , a response strongly inhibited by the ROS scavenger N-acetyl-L-cysteine (NAC) or the NADPH oxidase (NOX) inhibitor GKT137831. These inhibitors also affected CASP1 and NLRP3 mRNA and protein expression. More importantly, the serum/glucocorticoid regulated kinase 1 (SGK1) inhibitor GSK650394, or its shRNAs, completely abrogated the IL-1β mRNA response to Cl– and the IL-1β secretion, interrupting the autocrine IL-1β loop. The results suggest that Cl– effects are mediated by SGK1, in which under Cl– modulation stimulates the secretion of mature IL-1β, in turn, responsible for the upregulation of ROS, CASP1, NLRP3 and IL-1β itself, through autocrine signalling.

Bibliographic Details

Main Authors:	Clauzure, Mariángeles, Valdivieso, Ángel Gabriel, Dugour, Andrea V., Mori, Consuelo, Massip Copiz, María Macarena, Aguilar, María de los Angeles, Sotomayor, Verónica, Asensio, Cristian J. A., Figueroa, Juan M., Santa Coloma, Tomás Antonio
Format:	Artículo biblioteca
Language:	eng
Published:	Wiley-Blackwell 2021
Subjects:	FIBROSIS QUISTICA REGULADOR DE LA CONDUCTANCIA TRANSMEMBRANA, REGULADOR DE CONDUCTANCIA TRANSMEMBRANA DE LA FIBROSIS QUÍSTICA, SEÑALIZACION AUTOCRINA, ANIÓN CLORURO, CANAL DE CLORURO, INFLAMASOMA, INTERLEUCINA, NLRP3, SEGUNDO MENSAJERO, SGK1,
Online Access:	https://repositorio.uca.edu.ar/handle/123456789/14599
Tags:	Add Tag No Tags, Be the first to tag this record!