Assessment of a potential application of endogenous stem cells to treat congenital disorders

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Congenital diseases often result in incomplete organogenesis due to impaired stem cells function caused by toxic effects of gene mutations. Although surgical interventions are the primary treatment for these organ defects, their effectiveness remains unsatisfactory. However, advancements in genome editing technology now enable the in vivo correction of gene mutations in stem cells. To assess the potential of diseased stem cells to restore their ability to form autonomous tissues through gene repair, we successfully developed a mouse model of Hirschsprung disease (HSCR), serving as a model for congenital stem cell disorders. This model facilitates the removal of the disease-causing mutation through Cre-loxP-mediated gene recombination. The heterozygous mouse exhibited enteric nervous system defects resembling HSCR pathology. By crossing this mouse with Phox2b-Cre mice, we confirmed that deleting RET S811F in vagal neural crest-derived ENS stem cells evaded the enteric nerve defect phenotype. Moreover, crossing the HSCR mouse with Dhh-Cre mice eliminated the dominant negative mutation in Schwann cell precursors, which crucial for postnatal intestinal neurogenesis. We observed an increase in mCherry-positive intestinal neurons in regions lacking proper nerve function, suggesting compensation by cells where dominant-negative mutations were removed. The analysis of removing the disease-causing mutation in vagal neural crest-derived cells at different developmental time periods (Embryonic day 10.5. E11.5, and E12.5) revealed no notable differences among the observed time periods.