Deriving human ENS lineages for cell therapy and drug discovery in Hirschsprung disease

Abstract

The enteric nervous system (ENS) is the largest component of the autonomic nervous system with neuron numbers surpassing those present in the spinal cord1. The ENS has been called the “second brain”1 given its autonomy, remarkable neurotransmitter diversity and complex cytoarchitecture. Defects in ENS development are responsible for many human disorders including Hirschsprung's disease (HSCR). HSCR is a caused by the developmental failure of ENS progenitors to migrate into the GI tract in particular the distal colon2. Human ENS development remains poorly understood due to the lack of an easily accessible model system.Here we demonstrate the efficient derivation and isolation of ENS progenitors from human pluripotent stem cells (hPSCs) and their further differentiation into functional enteric neurons. In vitro derived ENS precursors are capable of targeted migration in the developing chick embryo and extensive colonization of the adult mouse colon. In vivo engraftment and migration of hPSC-derived ENS precursors rescues disease-related mortality in HSCR mice (EDNRBs-l/s-l), though mechanism of action remains unclear. Finally, EDNRB null mutant ENS precursors enable modeling of HSCR-related migration defects and the identification of Pepstatin A as candidate therapeutics. Our study establishes the first hPSC-based platform for the study of human ENS development and presents cell and drug-based strategies for the treatment of HSCR.

A Modular Platform for Differentiation of Human PSCs into All Major Ectodermal Lineages

“…The ability of generating regional specific CNS lineages is well known in the field and similar strategies are currently under development for generating specific NC lineages such as the switch from the default cranial to vagal and enteric NC (Fattahi et al, 2016). The final step in a human ectodermal lineage project will be the induction, isolation and molecular characterization of the various neural and non-neuronal subtypes generated from each region specific ectodermal lineage.…”

White paper on guidelines concerning enteric nervous system stem cell therapy for enteric neuropathies

“…Here we identify the main candidate stem/progenitor cell populations and highlight the respective pros and cons of each cell type in the context of enteric neuronal replacement. Of note is an exciting study recently published by Studer and colleagues (Fattahi et al, 2016). These authors directed human embryonic stem (ES) cells towards a vagal neural crest (i.e.…”

Human induced pluripotent stem cells for modelling neurodevelopmental disorders

“…Such hiPSC-based drug discovery systems are also already well on their way. In some cases, abnormal neurobiological phenotypes have already been reversed with existing drugs 74,176,177 . It is likely that the predictive validity of this system will only increase as the neurons used become more sophisticated and more accurately representative of human neurons.…”