The −172 A-to-G variation in ADAM17 gene promoter region affects EGR1/ADAM17 pathway and confers susceptibility to septic mortality with sepsis-3.0 criteria

• Published in: International immunopharmacology 2022-01, Vol.102, p.108385-108385, Article 108385
• Main Authors: He, Junbing, Zhao, Tian, Liu, Lizhen, Liao, Shuanglin, Yang, Shuai, Lu, Furong, Hong, Yuan, Wei, Ning, Cheng, Hongxiao, Zhang, Wenying, Shao, Yiming
• Format: Article
• Language: English
• Subjects: ADAM protein; ADAM17; ADAM17 Protein - genetics; ADAM17 Protein - immunology; Adult; Aged; Alleles; Animals; Assaying; Binding; Bioinformatics; Chromatin; Cytokines; Cytokines - blood; Cytokines - immunology; Early Growth Response Protein 1 - genetics; Early Growth Response Protein 1 - immunology; EGR-1 protein; EGR1; Endotoxemia; Enzyme-linked immunosorbent assay; Female; Gene polymorphism; Genotypes; Human Umbilical Vein Endothelial Cells; Humans; Immunoprecipitation; Inflammation; Inflammatory response; Kaplan-Meier Estimate; Leukocytes, Mononuclear - metabolism; Lipopolysaccharides; Male; Medical prognosis; Mice; Mice, Inbred C57BL; Middle Aged; Mortality; Patients; Polymorphism; Polymorphism, Single Nucleotide; Prognosis; Promoter Regions, Genetic; Proteolysis; RAW 264.7 Cells; Sepsis; Sepsis - genetics; Sepsis - immunology; Sepsis - mortality; Survival; Survival analysis
• ISSN: 1567-5769; 1878-1705
• DOI: 10.1016/j.intimp.2021.108385

•ADAM17 −172A > G polymorphism conferred susceptibility to septic mortality.•ADAM17 −172A > G polymorphism enhanced ADAM17 expression by the EGR1/ADAM17 pathway.•Inhibition of EGR1 attenuated ADAM17 expression and septic response.•Our findings provided new targets for sepsis risk assessment and prevention. A disintegrin and metalloproteinase 17 (ADAM17) is a proteolytic cleaving protein with a crucial function in the inflammatory responses, especially sepsis. But the clear role of ADAM17 in sepsis and the underlying mechanism remained unknown. In this study, we aim to determine the clinical association of ADAM17 −172A > G (rs12692386) promoter polymorphism with sepsis and to further explore the effect and mechanism of the early growth response 1 (EGR1)/ADAM17 pathway in inflammatory process during sepsis. A total of 477 sepsis patients and 750 controls were enrolled in this study to determine the association of ADAM17 −172A > G polymorphism with sepsis. The transcription factor binding to the promoter region of ADAM17 gene was predicted by bioinformatics analysis and verified by Chromatin Immunoprecipitation (ChIP) and luciferase assays. Quantitative real-time PCR and Western blot were performed to detect EGR1 and ADAM17 expression. Cytokine production was detected by enzyme-linked immunosorbent assay. The effect of EGR1/ADAM17 pathway on sepsis-induced inflammatory responses was evaluated in EGR1-silenced cells and endotoxemia mouse model. The frequencies of non-survivors among the sepsis patients with the −172AG/GG genotypes and G allele were distinctly higher than those among patients with the AA genotype (53.9% vs. 39.7%, OR = 1.779, 95% CI = 1.119–2.829, P = 0.0142) and A allele (30.9% vs. 22.2%, OR = 1.570, 95% CI = 1.095–2.251, P = 0.0136). The Kaplan-Meier survival analysis indicated that the 28-day survival in septic patients with −172AG/GG genotypes of this functional ADAM17 promoter polymorphism was much worse than in the AA genotype carriers (log-rank = 5.358, P = 0.021). The results of in vitro lipopolysaccharide-stimulated and luciferase assays indicated that the −172 A-to-G variation could functionally upregulate promoter activity and transcription of ADAM17 gene via enhancing the binding affinity of its promoter region with the EGR1. The ChIP assay identified the direct interaction. Further studies demonstrated that inhibition of EGR1 significantly decreased ADAM17 expression and the pro-inflammatory cytokine secretion in vitro, and improve