Notch signalling governs human enteric nervous system progenitor dynamics

The enteric nervous system (ENS) is the main branch of the peripheral nervous system that innervates the gastrointestinal tract controlling vital functions. It arises during embryogenesis via migration and differentiation of neural crest-derived ENS progenitors. Perturbation of these processes, caused by mutations in key signalling pathway components and transcription factors, prevents progenitor colonisation of the distal gut causing aganglionic phenotypes and enteric neuropathies such as Hirschsprung (HSCR) disease. While animal models implicate Notch signalling in ENS specification, its role in human ENS progenitor cell fate decisions remains unclear. Here, we employ a human pluripotent stem cell-based model to show that Notch signalling regulates the tempo of ENS progenitor differentiation. Quantitative modelling of our in vitro data supports a branching lineage model marked by an early pro-neurogenic bias; Notch signalling attenuation accelerates differentiation coincident with a shift toward increased gliogenesis. Furthermore, we establish that Notch signalling influences human ENS progenitor migration. Together, these findings provide mechanistic insights into how Notch signalling disruption may contribute to the pathogenesis of human intestinal aganglionosis.