Narcolepsy type 1 (NT1) is a chronic neurological disorder with a genetic predisposition, yet its precise molecular pathogenesis remains unclear. Despite increasing recognition of its clinical and genetic basis, the precise molecular mechanisms remain unclear. This study aimed to identify NT1-associated hub genes and investigate their biological functions and interactions through comprehensive bioinformatics analysis, followed by experimental validation.

NT1-related microarray data (GSE21592) were retrieved from Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified using integrated analysis with R software and the GEO2R platform. Functional enrichment analyses for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were conducted using Database for Annotation, Visualization, and Integrated Discovery online tool. Protein–protein interaction (PPI) networks were constructed using STRING database and visualized with Cytoscape. Quantitative real-time PCR (qRT-PCR) was used to experimentally validate the expression of identified hub genes in NT1 patients.

A total of 148 DEGs were identified. GO analysis revealed involvement primarily in biosynthesis, humoral immune response, viral gene expression, oxidoreductase activity, cytoplasmic translation, etc. KEGG analysis showed enrichment in oxidative phosphorylation, ribosomal pathways, thermogenesis, and glutathione metabolism pathways. Five hub genes – CREB1, PIK3R1, MED1, GATA3, and KDM5A – were identified from the PPI network. qRT-PCR validation confirmed significantly reduced expression of these genes in NT1 patients compared to healthy controls.

Our study identified and experimentally validated five critical hub genes associated with NT1, providing new insights into its molecular mechanisms and highlighting potential therapeutic targets for future research and clinical intervention strategies.