Enteric mesenchymal cells support the growth of postnatal enteric neural stem cells
pubmed: wnt1 2021-05-07
Stem Cells. 2021 May 2. doi: 10.1002/stem.3388. Online ahead of print.
Interplay between embryonic enteric neural stem cells (ENSCs) and enteric mesenchymal cells (EMCs) in the embryonic gut is essential for normal development of the enteric nervous system. Disruption of these interactions underlies the pathogenesis of intestinal aganglionosis in Hirschsprung disease (HSCR). ENSC therapy has been proposed as a possible treatment for HSCR, but whether the survival and development of postnatal-derived ENSCs similarly rely on signals from the mesenchymal environment is unknown and has important implications for developing protocols to expand ENSCs for cell transplantation therapy. Enteric neural crest-derived cells (ENCDCs) and EMCs were cultured from the small intestine of Wnt1-Rosa26-tdTomato mice. EMCs promoted the expansion of ENCDCs 9.5-fold by inducing ENSC properties, including expression of Nes, Sox10, Sox2, and Ngfr. EMCs enhanced the neurosphere-forming ability of ENCDCs, and this persisted after withdrawal of the EMCs. These effects were mediated by paracrine factors and several ligands known to support neural stem cells were identified in EMCs. Using the optimized expansion procedures, neurospheres were generated from small intestine of the Ednrb-/- mouse model of HSCR. These ENSCs had similar proliferative and migratory capacity to Ednrb+/+ ENSCs, albeit neurospheres contained fewer neurons. ENSCs derived from Ednrb-/- mice generated functional neurons with similar calcium responses to Ednrb+/+ ENSCs and survived after transplantation into the aganglionic colon of Ednrb-/- recipients. EMCs act as supporting cells to ENSCs postnatally via an array of synergistically acting paracrine signaling factors. These properties can be leveraged to expand autologous ENSCs from patients with HSCR mutations for therapeutic application. © AlphaMed Press 2021 SIGNIFICANCE STATEMENT: Hirschsprung disease (HSCR) is a congenital disorder characterized by failure of the nervous system to develop in the most distal part of intestine. It is known that mesenchymal cells play an important role for this process. Surgical removal is the only available treatment for HSCR, but outcomes are often unsatisfactory. Cell-based therapy offers potential to preserve colorectal function. Here, intestinal mesenchymal cells are identified to act as supporting cells to enteric neural stem cells (ENSCs) postnatally. This phenomenon was leveraged to develop innovative protocols to expand ENSCs from mice with HSCR, bringing cell therapies closer to therapeutic application.