Enteric neural stem cell transplant restores gut motility in mice with Hirschsprung disease
Ahmed A. Rahman, … , Allan M. Goldstein, Ryo Hotta
Ahmed A. Rahman, … , Allan M. Goldstein, Ryo Hotta
Published July 23, 2024
Citation Information: JCI Insight. 2024;9(17):e179755. https://doi.org/10.1172/jci.insight.179755.
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Research Article Cell biology Gastroenterology

Enteric neural stem cell transplant restores gut motility in mice with Hirschsprung disease

Abstract

The goal of this study was to determine if transplantation of enteric neural stem cells (ENSCs) can rescue the enteric nervous system, restore gut motility, reduce colonic inflammation, and improve survival in the Ednrb-KO mouse model of Hirschsprung disease (HSCR). ENSCs were isolated from mouse intestine, expanded to form neurospheres, and microinjected into the colons of recipient Ednrb-KO mice. Transplanted ENSCs were identified in recipient colons as cell clusters in “neo-ganglia.” Immunohistochemical evaluation demonstrated extensive cell migration away from the sites of cell delivery and across the muscle layers. Electrical field stimulation and optogenetics showed significantly enhanced contractile activity of aganglionic colonic smooth muscle following ENSC transplantation and confirmed functional neuromuscular integration of the transplanted ENSC-derived neurons. ENSC injection also partially restored the colonic migrating motor complex. Histological examination revealed a significant reduction in inflammation in ENSC-transplanted aganglionic recipient colon compared with that of sham-operated mice. Interestingly, mice that received cell transplant also had prolonged survival compared with controls. This study demonstrates that ENSC transplantation can improve outcomes in HSCR by restoring gut motility and reducing the severity of Hirschsprung-associated enterocolitis, the leading cause of death in human HSCR.

Authors

Ahmed A. Rahman, Takahiro Ohkura, Sukhada Bhave, Weikang Pan, Kensuke Ohishi, Leah Ott, Christopher Han, Abigail Leavitt, Rhian Stavely, Alan J. Burns, Allan M. Goldstein, Ryo Hotta

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Figure 6

ENSC transplantation restores colonic motility in mice with HSCR and prolongs their survival.

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ENSC transplantation restores colonic motility in mice with HSCR and pro...
Representative spatiotemporal map kymographs generated from video recordings of colonic motility from Ednrb WT (n = 6), Ednrb-KO (n = 5), and Ednrb-KO + cells (n = 3) mice 2 weeks after cell transplant, depicting colonic contraction (red) and relaxation (yellow) along the length of the colon over time. The propagating CMMCs observed in WT mice are absent in KO mice but are partially restored following cell transplantation (Ednrb-KO + Cells) (A). Simultaneous intraluminal pressure recordings show effective colorectal contractility in WT mice, minimal pressure generation in Ednrb-KO mice, and significant restoration after cell transplant (B and F). CMMC frequency (C), velocity (D), and distance propagated (E) are all markedly increased in the Ednrb-KO + Cells group compared with the Ednrb-KO group. (G) Survival curve of Ednrb-KO mice that underwent ENSC transplantation (n = 3) or no treatment (n = 5). Statistical significance was determined by log-rank (Mantel-Cox) test (G). All the values represent the mean of 2–4 animals for each group, repeated 2–3 times. Data are shown as the mean ± SEM. Statistical significance was determined by the 1-way ANOVA with a post hoc Tukey’s test (C–F). *P < 0.05, **P < 0.01, and ***P < 0.001 are statistically significant.