Defective OXPHOS perturbs the neuronal differentiation of human enteric neural crest cells as revealed by the iPSC model of Hirschsprung disease and single-cell transcriptomic analysis [Poster presentation]

Abstract

<p>Hirschsprung disease (HSCR) is characterized by the absence of enteric neurons caused by the defects of enteric neural crest cells (ENCCs), leading to intestinal obstruction. Here, using iPSC-based models of HSCR and single-cell transcriptomic analysis, we identified a gene set of 118 genes commonly dysregulated in all HSCR-ENCCs, with <em>HDAC1</em> found to be a key regulator of these genes. Furthermore, upregulation of RNA splicing mediators and enhanced alternative splicing events were associated with severe form of HSCR. In particular, the higher inclusion rate of exon 9 in <em>PTBP1</em> and the perturbed expression of a PTBP1-target, <em>PKM, </em>were significantly enriched in these HSCR-ENCCs, and associated with the defective oxidative-phosphorylation (OXPHOS) and impaired neurogenesis. Inhibition of HDAC1 and up-regulation of <em>PKM1</em> significantly enhanced the differentiation capacity of HSCR-ENCCs. In sum, we have defined the core factors underpinning HSCR disease and demonstrated the implications of OXPHOS in ENCC development and HSCR pathogenesis.</p>