Analysis and verification of potential ferroptosis-related diagnostic markers in the early stage of metabolic dysfunction-associated steatohepatitis.

Metabolic dysfunction-associated steatohepatitis (MASH) is currently the fastest-growing cause of liver-related deaths. Intervention in the early stage of MASH will effectively reverse the condition and avoid huge health and economic burdens. To date, our understanding of MASH remains quite limited. Its pathogenesis remains not fully understood, and effective diagnostic and therapeutic tools are lacking. Although ferroptosis is closely related to MASH and seen as a promising target, its exact mechanism in the early stage of MASH remains unclear. This study aims to analyze the ferroptosis-related genes that are differentially expressed in early MASH, identify potential diagnostic markers, and provide a new perspective for further in-depth research on the role of ferroptosis in early MASH. Gene expression datasets of human MASH patients were obtained from the GEO database. The early MASH gene expression data from GSE126848 and GSE162694 were analyzed. Genes with significant P-values were subjected to GO enrichment analysis. After intersecting the obtained genes with those in the Ferroptosis Database (FerrDb), ferroptosis signature genes in early MASH were screened and then analyzed via PPI network. Four hub genes (FXN, FABP4, QSOX1, CAPG) were selected and validated in the GSE135251 dataset. Their diagnostic efficiency was evaluated using ROC curve analysis. In vitro and in vivo MASH models, including HFCFD-induced mouse models and FFA-induced mouse liver cells, confirmed their expression changes related to ferroptosis. Also, in human fibrotic liver tissues, three of these hub genes were highly expressed in macrophages. These four genes may serve as potential diagnostic biomarkers for early MASH.