Inhibition of chondroitin sulfate proteoglycans enhances cell migration following enteric neural stem cell...

Enteric nervous system development relies on migration, proliferation and differentiation of enteric neural crest-derived cells (ENCDCs), processes that are regulated by mesenchymal morphogens and the extracellular matrix (ECM). Hirschsprung disease (HD) is a congenital neurocristopathy in which abnormal gut motility results from loss of ENCDC-derived enteric ganglia in the colorectum. While enteric neural stem cell transplantation is a proposed therapy, current efforts are limited by poor cell migration in HD models. To address this, we investigated ECM-derived inhibitory cues on enteric neural stem cell transplantation efficiency. Immunohistochemistry of embryonic chick hindgut reveals dynamic chondroitin sulfate proteoglycan (CSPG) expression during development. In vitro assays show that CSPGs inhibit ENCDC migration, an effect reversed by chondroitinase ABC. Transplantation of enteric neurospheres into chick and mouse colon results in limited engraftment, but chondroitinase ABC significantly enhances migration of transplanted cells. Single-cell RNA sequencing of postnatal mouse neurospheres reveals high expression of CD44, a transmembrane glycoprotein that can act as a functional CSPG on the cell surface. Blocking CD44 function promotes neurosphere-derived ENCDC migration. Modulating inhibitory ECM components may enhance the success of regenerative therapies for HD.