ChIP-Seq-Based Approach in Mouse Enteric Precursor Cells Reveals New Potential Genes with a Role in Enteric Nervous System Development and Hirschsprung Disease.

Leticia Villalba-Benito,Raquel María Fernández,Ana Torroglosa,María José Moya-Jiménez,Salud Borrego,Berta Luzón-Toro,Guillermo Antiñolo

doi:10.3390/ijms21239061

Abstract

Hirschsprung disease (HSCR) is a neurocristopathy characterized by intestinal aganglionosis which is attributed to a failure in neural crest cell (NCC) development during the embryonic stage. The colonization of the intestine by NCCs is a process finely controlled by a wide and complex gene regulatory system. Several genes have been associated with HSCR, but many aspects still remain poorly understood. The present study is focused on deciphering the PAX6 interaction network during enteric nervous system (ENS) formation. A combined experimental and computational approach was performed to identify PAX6 direct targets, as well as gene networks shared among such targets as potential susceptibility factors for HSCR. As a result, genes related to PAX6 either directly (RABGGTB and BRD3) or indirectly (TGFB1, HRAS, and GRB2) were identified as putative genes associated with HSCR. Interestingly, GRB2 is involved in the RET/GDNF/GFRA1 signaling pathway, one of the main pathways implicated in the disease. Our findings represent a new contribution to advance in the knowledge of the genetic basis of HSCR. The investigation of the role of these genes could help to elucidate their implication in HSCR onset.

Highlights

Hirschsprung disease (HSCR, OMIM 142623) or aganglionic megacolon, is a disorder characterized by intestinal aganglionosis in a variable segment of the distal gut, whose main manifestation is constipation or intestinal obstruction [1]
enteric precursor cells (EPCs) grow in culture as cell aggregates known as neurosphere-like bodies (NLBs), which are a population of neural crest-derived multipotent stem cells present in postnatal gut
The current study was focused on the identification of PAX6 gene regulatory networks during enteric nervous system (ENS) development to detect additional genes and mechanisms to better outline HSCR etiology

Summary

Introduction

Hirschsprung disease (HSCR, OMIM 142623) or aganglionic megacolon, is a disorder characterized by intestinal aganglionosis in a variable segment of the distal gut, whose main manifestation is constipation or intestinal obstruction [1]. A large number of different genes have been related to HSCR. In most cases, these genes encode molecules that are widely involved in the major signaling pathways associated with HSCR, the most important being the RET/GDNF/GFRA1 and EDN3/EDNRB signaling pathways [4]. The genetic cause of the disease in a large percentage of HSCR patients remains unknown; the search for additional genes and mechanisms to better outline HSCR etiology is still required. HSCR, especially the short segment forms, is a disorder with a complex genetic basis that could be explained by the heterogeneity of ENS development, which is regulated by an ever-increasing range of molecules. Despite the fact that some families show an either dominant or recessive pattern, most of the cases show a complex and non-Mendelian mode of inheritance with variable penetrance [2,5]

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