The Role of Neutrophil Extracellular Traps (NETs) in Non-alcoholic Fatty Liver Disease (NAFLD): A Comprehensive Analysis of NETs-related Genes

Abstract

Abstract Non-alcoholic Fatty Liver Disease (NAFLD), prevalent among adults, has become a dominant chronic liver condition worldwide, with a rising incidence of liver cirrhosis. The progression of NAFLD is critically influenced by Neutrophil Extracellular Traps (NETs), which play a key role in its pathogenesis. However, the specific functions of NETs-related genes within NAFLD necessitate further in-depth research. Our team utilized advanced methodologies including AddModuleScore, ssGSEA, and WGCNA for gene screening, identifying NETs-linked genes in single-cell and bulk transcriptomic data. Through algorithms such as Random Forest, Support Vector Machine, Least Absolute Shrinkage and Selection Operator, and Selector Operator, we identified ZFP36L2 and PHLDA1 as significant hub genes. Their role in NAFLD diagnosis was validated using the training dataset GSE164760 and further confirmed in an animal model. The study pinpointed 116 NET-associated genes, predominantly involved in immune and metabolic pathways. Notably, PHLDA1 and ZFP36L2 were determined as hub genes via machine learning techniques, contributing to a predictive model. These genes are involved in inflammatory and metabolic processes, with single-cell RNA sequencing (scRNA-seq) revealing distinct cellular communication patterns based on their expression. In conclusion, this research elucidates the molecular characteristics of NET-associated genes in NAFLD, identifying PHLDA1 and ZFP36L2 as potential biomarkers. By exploring their roles in the hepatic microenvironment, our findings offer significant insights for diagnosing and managing NAFLD, ultimately aiming to enhance patient outcomes.