Regulatory gene analysis in enteric neural crest cells stratified by GDNF activation status: An insight into...

Incomplete migration of enteric neural crest cells (ENCCs) into the colon underlies the pathogenesis of Hirschsprung disease, resulting in aganglionosis and disordered intestinal motility. Although activation of the glial cell line-derived neurotrophic factor (GDNF) pathway has been implicated in neurosphere differentiation, specific regulatory genes governing ENCC differentiation and proliferation remain poorly characterized. This study seeks to elucidate regulatory genes and pathways involved in neural crest cell differentiation, furthering our understanding of GDNF's role in enteric nervous system development.

Mus musculus (mice) ENCC gene expression data were obtained from Gene Expression Omnibus (GEO), a public functional genomics repository, to compare GDNF (n = 13) and non-GDNF (n = 12) samples in dataset GSE34208. We calculated repressive tendency scores using TRIAGE R to rank genes based on their typical epigenetic repression and identify unique regulatory genes in GDNF+ and GDNF- neurospheres. Regulatory pathways were then characterized using Gene Ontology (GO) database.

TRIAGE-based prioritization revealed greater similarity among GDNF + samples than between GDNF+ and GDNF- samples. GO enrichment analysis of 300 top-ranked genes from GDNF + samples demonstrated significant enrichment in biological processes relevant to ENS development. Homeobox (HOX) transcription factors emerged as a key regulatory gene class in the GDNF + condition, appearing consistently across pathways related to gut patterning and neuronal differentiation.

Our analysis clarifies the regulatory mechanisms governing neural crest cell differentiation and proliferation and establishes a foundation for future research into the molecular regulation of enteric neural crest cell development and Hirschsprung disease pathophysiology.

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