YTHDF1 alleviates sepsis by upregulating WWP1 to induce NLRP3 ubiquitination and inhibit caspase-1-dependent pyroptosis

Abstract

Pyroptosis is inflammation-associated caspase-1-dependent programmed cell death, which confers a crucial role in sepsis. The present study intends to investigate the regulatory network and function of the microarray-predicted YTHDF1 in caspase-1-dependent pyroptosis of sepsis. Peripheral blood of patients with sepsis was collected to determine WWP1 and YTHDF1 expression. An in vitro sepsis cell model was induced in RAW264.7 cells using lipopolysaccharide (LPS) and ATP and an in vivo septic mouse model by cecal ligation and perforation (CLP). After gain- and loss-of-function assays in vitro and in vivo, TNF-α and IL-1β levels and the cleavage of gasdermin-D (GSDMD) were detected by ELISA and Western blot assay, followed by determination of lactate dehydrogenase (LDH) activity. Immunoprecipitation and meRIP assay were performed to detect the ubiquitination of NLRP3 and the m6A modification of WWP1 mRNA. The binding of WWP1 to YTHDF1 was explored using RIP-RT-qPCR and dual luciferase gene reporter assay. It was noted that WWP1 and YTHDF1 were downregulated in clinical sepsis samples, LPS + ATP-treated RAW264.7 cells, and CLP-induced mice. The ubiquitination of NLRP3 was promoted after overexpression of WWP1. WWP1 translation could be promoted by YTHDF1. Then, WWP1 or YTHDF1 overexpression diminished LDH activity, NLRP3 inflammasomes and caspase-1-mediated cleavage of GSDMD in LPS + ATP-induced RAW264.7 cells. Overexpressed YTHDF1 restrained inflammatory response in CLP-induced mice. Collectively, the alleviatory effect of m6A reader protein YTHDF1 may be achieved through promotion of NLRP3 ubiquitination and inhibition of caspase-1-dependent pyroptosis by upregulating WWP1.

USP9X promotes lipopolysaccharide‐stimulated acute lung injury by deubiquitination of NLRP3

“…Moreover, Enteropathogenic Escherichia coli uses NleA to inhibit NLRP3 inflammasome activation by ubiquitination of NLRP3 (Yen et al, 2015), and LPS along with ATP stimulation initiated K63-and K48-linked polyubiquitination of NLRP3 (J. Tang et al, 2020;Zhang et al, 2022). In the present study, we found that increased K48-linked polyubiquitination of NLRP3 was observed upon downregulating USP9X.…”

“…Previous studies showed that hepatitis C virus infection inhibited K63‐linked polyubiquitination of NLRP3 (Ramachandran et al, 2021) and Giardia infection‐induced NLRP3 inflammasome activation was involved in its K63 deubiquitination (L. Liu et al, 2021). Moreover, Enteropathogenic Escherichia coli uses NleA to inhibit NLRP3 inflammasome activation by ubiquitination of NLRP3 (Yen et al, 2015), and LPS along with ATP stimulation initiated K63‐ and K48‐linked polyubiquitination of NLRP3 (J. Tang et al, 2020; Zhang et al, 2022). In the present study, we found that increased K48‐linked polyubiquitination of NLRP3 was observed upon downregulating USP9X.…”

Silencing M6a Reader Ythdc1 Reduces Inflammatory Response in Sepsis-Induced Cardiomyopathy by Inhibiting Serpina3n Expression

“…Among many epigenetic modifications studied, the m 6 A modification in RNA has been proven to play an important role in the development and treatment of sepsis. However, the specific function of m 6 A “reading” protein in SIC inflammatory reaction is unclear and whether m 6 A is involved in this process remains unknown (14,41–43). In this study, the m 6 A reader, YTHDC1, was found to increase after LPS stimulation.…”

The Role of Post-Translational Modifications in Regulation of NLRP3 Inflammasome Activation

“…Recent advances in the development of pharmacological targeting of ubiquitination and deubiquitination have uncovered a great potential for treatments of cancer, neurodegenerative disorders, inflammatory disorders, immunological diseases and microbial infection [ 30 ]. YTH N6-methyladenosine (m6A) RNA-binding protein 1 (YTHDF1), a reader of m6A, alleviates cecal ligation and perforation-induced sepsis through promoting NLRP3 ubiquitination [ 31 ]. Tranilast blunts NLRP3 inflammasome assembly via enhancing NLRP3 ubiquitination, contributing to the amelioration of vascular inflammation and atherosclerosis [ 32 ].…”