Role of cholinergic neuroimmune signaling pathway disruption in the pathogenesis of hirschsprung-associated enterocolitis

Federal Project Grant Award Summary

Massachusetts General Hospital received a $439,216 Project Grant award dated July 8, 2025, from the National Institute of Allergy and Infectious Diseases under the Allergy and Infectious Diseases Research program (CFDA 93.855) to conduct a two-year research initiative examining the role of cholinergic neuroimmune signaling pathway disruption in Hirschsprung-associated enterocolitis (HAEC). The research will characterize neuroimmune mechanisms underlying HAEC, a devastating bowel inflammatory disorder affecting children with Hirschsprung disease, and test cell-based therapeutic strategies to restore immune homeostasis and improve epithelial barrier function. The project will utilize murine models, cell and organoid co-culture technologies, optogenetics, and cell transplantation techniques to investigate how loss of the enteric nervous system disrupts innate lymphoid cell and tuft cell interactions, ultimately leading to intestinal inflammation and immune dysregulation.

The anticipated deliverables include scientific findings regarding the functional relationships between enteric neuronal cells, immune cells, and epithelial architecture in the context of HAEC pathophysiology, as well as proof-of-concept data supporting enteric neuronal cell therapy and optogenetic interventions as potential therapeutic approaches. These research outputs will advance understanding of neuroimmune interactions in gastrointestinal inflammatory disorders and may inform treatment strategies to reduce morbidity and mortality in pediatric patients with Hirschsprung disease. The project period extends through July 7, 2027, with performance conducted at the award recipient's Research Management Division in Boston, Massachusetts.

Abstract

Hirschsprung disease (hscr) is a congenital disorder of gastrointestinal tract characterized by the absence of enteric nervous system (ENS) along variable lengths of distal intestine. Children with hscr are at risk for a devastating bowel inflammatory disorder, called hirschsprung- associated enterocolitis (haec), that represents the leading cause of long-term morbidity and mortality and is associated with significant healthcare costs. Gaps in therapy are due to our incomplete understanding of the molecular and cellular causes of this condition. While lack of the ENS and immune dysregulation have been implicated in the pathophysiology of the haec, little is known about the role of neuroimmune interactions, which are so important in multiple functional gastrointestinal diseases. Our broad objective is to characterize the neuroimmune mechanisms underlying haec and to leverage this knowledge to test cell-based strategies to restore immune homeostasis and improve epithelial barrier function and the GUT immunity. Based on our preliminary data, we hypothesize that disruption of normal ENS-innate lymphoid cell (ILC)-tuft cell (TC) interactions leads to the development and progression of intestinal inflammation. By utilizing murine model of hscr, cell and organoid CO-culture technologies, optogenetics, and cell transplantation techniques, we propose to test our hypothesis that loss of ENS in the aganglionic bowel of hscr leads to disfunction of ILC-TC circuit that is associated with alterations of epithelial architecture and immune cell infiltration and disrupted intestinal innate immunity. We will also test our hypothesis that enteric neuronal cell therapy and optogenetic approach can restore immune homeostasis in vivo. The proposed studies will enhance our understanding of the role of the ENS in the immune dysregulation and marked declines in ILC and TC populations observed in the haec, and the results will allow US to identify novel potential therapies that target the primary cause of haec pathophysiology.