Abstract
Objective
HSCR is caused by disruption of complex signaling pathways within the gene regulatory network (GRN) during enteric nervous system (ENS) development, including glial cell line-derived neurotrophic factor (GDNF) and GDNF family receptor alpha-1 (GFRα1). However, pathogenic variants in all GRN genes account for only ~ 80% cases; therefore, the epigenetic role remains to be elucidated. We compared GDNF and GFRα1 expression between HSCR patients and controls.Results
qPCR revealed an upregulated GDNF expression in both ganglionic (206.37-fold) and aganglionic (126.35-fold) HSCR compared to control colons (ΔCT 7.65 ± 2.19 vs. 15.34 ± 1.34; p = 0.0001; and ΔCT 8.35 ± 2.57 vs. 15.34 ± 1.34; p = 0.0001). qPCR also showed an upregulated GFRα1 expression in both ganglionic (29.66-fold) and aganglionic (18.44-fold) HSCR compared to control colons (ΔCT 8.44 ± 1.98 vs. 13.33 ± 1.36; p = 0.0001; and ΔCT 9.13 ± 1.46 vs. 13.33 ± 1.36; p = 0.0001). In addition, no significant differences were observed between ganglionic and aganglionic segments for either GDNF or GFRα1 (ΔCT 7.65 ± 2.19 vs. 8.35 ± 2.57; p = 0.43; and ΔCT 8.44 ± 1.98 vs. 9.13 ± 1.46; p = 0.29). Our study demonstrates global aberrant expression of GDNF and GFRα1 in HSCR patients. These findings underscore the complexity of HSCR as a multifactorial disorder and highlight the importance of integrated signaling networks in ENS development, providing a rationale for further mechanistic and translational studies.