Intramuscular Enteric Glia Persist in Hirschsprung Disease and Undergo Neurogenesis in Response to GDNF-NCAM1 Signaling

Background & Aims Enteric glial cells (EGCs) play vital roles in gut homeostasis and neurogenesis. Hirschsprung disease (HSCR) is a neurocristopathy, yet paradoxically, neural crest-derived enteric glial cells (EGCs) are present within the muscle of the affected region. This study investigates the molecular identity, origins, and neurogenic potential of EGCs in the aganglionic mouse and human colon. Methods We utilized single-cell RNA sequencing (scRNA-seq) from aganglionic and ganglionated segments of 10-14 day old Ednrb-null mice (Plp1-GFP;Ednrb-/-) and wild-type controls. Results scRNA-seq revealed the absence of GFAP+ intraganglionic (IG) glia in aganglionic colon, while CAMK2b+ extraganglionic (EG) glia and Schwann-like cells (SLCs) were present. EG glia exhibited a transcriptional profile similar to SLCs, suggesting a possible shared embryonic origin. Conclusions A subpopulation of extraganglionic EGCs, transcriptionally similar to SLCs, persists in the aganglionic segment of HSCR. Overall design: Use of 10X Genomics technology to analyze single-cell transcriptomics in single cell suspensions of digested tissue from aganglionic and ganglionic tissue from Plp1-GFP;Ednrb-/- mice.