Identification and Validation of Autophagy-Related Genes in Hirschsprung’s Disease

Abstract

Abstract Background Hirschsprung's disease (HSCR) is a congenital disorder characterized by aganglionosis in the intermuscular and submucosal nerve plexus of the intestines, resulting in impaired gastrointestinal function. The exact etiology and pathogenesis of HSCR are yet not fully understood. Accumulating evidence suggests that autophagy plays an important role in the pathogenesis of HSCR, but its specific mechanism needs to be further studied. Methods This study used the online Gene Expression Omnibus (GEO) microarray expression profiling datasets GSE96854 and GSE98502. R software was used to identify autophagy-related genes that displayed potential differential expression in HSCR. The differentially expressed autophagy-related genes were analyzed using correlation analysis, tissue-specific gene expression profiling, gene ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and protein-protein interaction (PPI) network analysis. Finally, HSCR specimens were utilized for in vitro verification of mRNA levels of key genes using quantitative real-time polymerase chain reaction (qRT-PCR). Results A total of 20 autophagy-related genes exhibiting differential expression were identified, comprising 15 up-regulated and five down-regulated genes. Enrichment analysis with the GO and KEGG pathways revealed significant enrichment in pathways associated with the regulation of autophagy, specifically macroautophagy. By conducting PPI network analysis and constructing key modules, we identified nine hub genes. Subsequently, the consistency between SIRT1 expression in the HSCR model and bioinformatics analysis of mRNA chip results was confirmed through qRT-PCR validation. Conclusion Through bioinformatics analysis, we identified 20 potential autophagy-related genes associated with HSCR. Among them, the upregulation of SIRT1 expression may impact the occurrence and progression of HSCR by regulating autophagy pathways. This provides a fresh outlook on the etiology of Hirschsprung's disease.