Expression of PROKR1 and PROKR2 in human enteric neural precursor cells and identification of sequence variants suggest a role in HSCR

PLoS One. 2011;6(8):e23475. doi: 10.1371/journal.pone.0023475. Epub 2011 Aug 12.

Abstract

Background: The enteric nervous system (ENS) is entirely derived from neural crest and its normal development is regulated by specific molecular pathways. Failure in complete ENS formation results in aganglionic gut conditions such as Hirschsprung's disease (HSCR). Recently, PROKR1 expression has been demonstrated in mouse enteric neural crest derived cells and Prok-1 was shown to work coordinately with GDNF in the development of the ENS.

Principal findings: In the present report, ENS progenitors were isolated and characterized from the ganglionic gut from children diagnosed with and without HSCR, and the expression of prokineticin receptors was examined. Immunocytochemical analysis of neurosphere-forming cells demonstrated that both PROKR1 and PROKR2 were present in human enteric neural crest cells. In addition, we also performed a mutational analysis of PROKR1, PROKR2, PROK1 and PROK2 genes in a cohort of HSCR patients, evaluating them for the first time as susceptibility genes for the disease. Several missense variants were detected, most of them affecting highly conserved amino acid residues of the protein and located in functional domains of both receptors, which suggests a possible deleterious effect in their biological function.

Conclusions: Our results suggest that not only PROKR1, but also PROKR2 might mediate a complementary signalling to the RET/GFRα1/GDNF pathway supporting proliferation/survival and differentiation of precursor cells during ENS development. These findings, together with the detection of sequence variants in PROKR1, PROK1 and PROKR2 genes associated to HSCR and, in some cases in combination with RET or GDNF mutations, provide the first evidence to consider them as susceptibility genes for HSCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Child, Preschool
  • Cohort Studies
  • Enteric Nervous System / cytology
  • Enteric Nervous System / metabolism
  • Female
  • Gastrointestinal Hormones / genetics
  • Gastrointestinal Hormones / metabolism
  • Gene Expression
  • Genetic Predisposition to Disease / genetics
  • Glial Cell Line-Derived Neurotrophic Factor / genetics
  • Glial Cell Line-Derived Neurotrophic Factor / metabolism
  • Glial Cell Line-Derived Neurotrophic Factor Receptors / genetics
  • Glial Cell Line-Derived Neurotrophic Factor Receptors / metabolism
  • Hirschsprung Disease / genetics*
  • Hirschsprung Disease / metabolism
  • Hirschsprung Disease / pathology
  • Humans
  • Immunohistochemistry
  • Infant
  • Infant, Newborn
  • Male
  • Microscopy, Confocal
  • Mutation*
  • Neural Stem Cells / metabolism*
  • Neuropeptides / genetics
  • Neuropeptides / metabolism
  • Proto-Oncogene Proteins c-ret / genetics
  • Proto-Oncogene Proteins c-ret / metabolism
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism
  • Receptors, Peptide / genetics*
  • Receptors, Peptide / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Vascular Endothelial Growth Factor, Endocrine-Gland-Derived / genetics
  • Vascular Endothelial Growth Factor, Endocrine-Gland-Derived / metabolism

Substances

  • GFRA1 protein, human
  • Gastrointestinal Hormones
  • Glial Cell Line-Derived Neurotrophic Factor
  • Glial Cell Line-Derived Neurotrophic Factor Receptors
  • Neuropeptides
  • PROK1 protein, human
  • PROK2 protein, human
  • PROKR1 protein, human
  • PROKR2 protein, human
  • Receptors, G-Protein-Coupled
  • Receptors, Peptide
  • Vascular Endothelial Growth Factor, Endocrine-Gland-Derived
  • Proto-Oncogene Proteins c-ret
  • RET protein, human