Single-cell WGCNA combined with transcriptome sequencing to study the molecular mechanisms of TRPM4-related genes in obesity with experimental validation

While TRPM4 is crucial in immune and metabolic diseases, its role in obesity remains unclear. This study aimed to identify TRPM4-associated key genes in obesity. Obesity-related transcriptomic data were analyzed using single-cell RNA sequencing (scRNA-seq) to identify critical cell types, high-Dimensional Weighted Gene Co-expression Network Analysis (hdWGCNA) to determine module genes, and machine learning to screen key genes. A diagnostic nomogram was constructed, potential therapeutics were predicted, and reverse transcription quantitative PCR (RT-qPCR) validated gene expression in clinical samples. Endothelial cells were identified as critical, with ETV6, LAPTM5, and MCOLN3 highlighted as key genes. The nomogram demonstrated strong diagnostic efficacy, and compounds including cyclosporin A, retinoic acid, arsenious acid, and trichostatin A were predicted as potential modulators. RT-qPCR confirmed elevated ETV6 and LAPTM5 and reduced MCOLN3 expression in obesity (P < 0.001 and P < 0.01, respectively). Integrating bulk and scRNA-seq analyses identified endothelial cells and genes ETV6, LAPTM5, and MCOLN3 as key players, offering potential targets for TRPM4-related obesity interventions.