Multi-omics Analysis Reveals Molecular Subtype of Mitochondrial Oxidative Stress and Prognostic Model Development in Sepsis

Background: Sepsis is a prevalent disease with high mortality involving severe systemic inflammatory responses. Although the mechanisms underlying sepsis have been widely explored, the occurrence and exacerbation of sepsis remain unclear, with limited therapeutic options. Inflammation and mitochondrial oxidative stress have been proposed as primary factors in the development of sepsis. Methods: In the present research, normal and sepsis samples were obtained from the Gene Expression Omnibus (GEO) database (GSE54514, GSE65682, and GSE95233). To identify the key mitochondrial oxidative stress-related gene (MOSRG) signature associated with sepsis, both weighted gene co-expression network analysis (WGCNA) and differential expression analysis were conducted. Least Absolute Shrinkage and Selection Operator (LASSO) analysis and univariate and multivariate Cox analysis were used to construct the prognostic risk model for sepsis. Immune infiltration characteristics were analyzed using the Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE) and single-sample Gene Set Enrichment Analysis (ssGSEA) algorithms. Single-cell RNA sequencing and in vitro experiments provided additional evidence for the pivotal role of RNA-binding protein, ribonuclease 2 (RNASE2) in the regulation of mitochondrial oxidative stress in sepsis. Results: Three MOSRGs RNASE2, CX3C chemokine receptor 1 (CX3CR1), and epoxide hydrolase 2 (EPHX2) were recognized as potential diagnostic indicators for sepsis in this study. The immune infiltration analysis provides strong evidence that three biomarkers were linked to immune-related mechanisms involved in the pathogenesis of sepsis. The pivotal role of RNASE2 in regulating mitochondrial oxidative stress during sepsis was confirmed using single-cell RNA-seq analysis and validated by in vitro molecular biology experiments. Inhibition of RNASE2 was found to significantly mitigate mitochondrial oxidative stress injury in sepsis. Conclusion: This research underscores the significant impact of mitochondrial oxidative stress-related genes on immune regulation in sepsis and highlights the potential therapeutic implications of candidate biomarkers.