Abstract
Genetic studies of Hirschsprung disease, a common congenital malformation, have identified eight genes with mutations that can be associated with this condition. Mutations at individual loci are, however, neither necessary nor sufficient to cause clinical disease. We conducted a genome-wide association study in 43 Mennonite family trios using 2,083 microsatellites and single-nucleotide polymorphisms and a new multipoint linkage disequilibrium method that searches for association arising from common ancestry. We identified susceptibility loci at 10q11, 13q22 and 16q23; the gene at 13q22 is EDNRB, encoding a G protein–coupled receptor (GPCR) and the gene at 10q11 is RET, encoding a receptor tyrosine kinase (RTK). Statistically significant joint transmission of RET and EDNRB alleles in affected individuals and non-complementation of aganglionosis in mouse intercrosses between Ret null and the Ednrb hypomorphic piebald allele are suggestive of epistasis between EDNRB and RET. Thus, genetic interaction between mutations in RET and EDNRB is an underlying mechanism for this complex disorder.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Puffenberger, E.G. et al. Identity-by-descent and association mapping of a recessive gene for Hirschsprung disease on human chromosome 13q22. Hum. Mol. Genet. 3, 1217–1225 (1994).
Puffenberger, E.G. et al. A missense mutation of the endothelin-B receptor gene in multigenic Hirschsprung's disease. Cell 79, 1257–1266 (1994).
Badner, J.A., Sieber, W.K., Garver, K.L. & Chakravarti, A. A genetic study of Hirschsprung disease. Am. J. Hum. Genet. 46, 568–580 (1990).
Chakravarti, A. & Lyonnet, S. Hirschsprung disease. in The Metabolic and Molecular Bases of Inherited Disease 8th edn (eds Scriver, C.R., Beaudet, A.L., Valle, D., Sly, W.S., Childs, B., Kinzler, K. & Vogelstein, B.) 6231–6255 (McGraw–Hill, New York, 2001).
Amiel, J. et al. Large-scale deletions and SMADIP1 truncating mutations in syndromic Hirschsprung disease with involvement of midline structures. Am. J. Hum. Genet. 69, 1370–1377 (2001).
Chakravarti, A. Endothelin receptor-mediated signaling in Hirschsprung disease. Hum. Mol. Genet. 5, 303–307 (1996).
Bergey, L.L. The early settlement of Waterloo Township, Ontario, Canada. Pa. Mennon. Herit. 15, 9–20 (1992).
Bolk, S. et al. A human model for multigenic inheritance: phenotypic expression in Hirschsprung disease requires both the RET gene and a new 9q31 locus. Proc. Natl Acad. Sci. USA 97, 268–273 (2000).
Gabriel, S.B. et al. Segregation at three loci explains familial and population risk in Hirschsprung disease. Nat. Genet. 31, 89–93 (2002).
Takeuchi, T. et al. Expression of T-cadherin (CDH13, H-Cadherin) in human brain and its characteristics as a negative growth regulator of epidermal growth factor in neuroblastoma cells. J. Neurochem. 74, 1489–1497 (2000).
Borrello, M.G. et al. The full oncogenic activity of Ret/ptc2 depends on tyrosine 539, a docking site for phospholipase C gamma. Mol. Cell. Biol. 16, 2151–2163 (1996).
Obermeier, A. et al. Neuronal differentiation signals are controlled by nerve growth factor receptor/Trk binding sites for SHC and PLC gamma. EMBO J. 13, 1585–1590 (1994).
Fitze, G., Schreiber, M., Kuhlisch, E., Schackert, H.K. & Roesner, D. Association of RET protooncogene codon 45 polymorphism with Hirschsprung disease. Am. J. Hum. Genet. 65, 1469–1473 (1999).
Borrego, S. et al. Specific polymorphisms in the RET proto-oncogene are over-represented in patients with Hirschsprung disease and may represent loci modifying phenotypic expression. J. Med. Genet. 36, 771–774 (1999).
Borrego, S. et al. RET genotypes comprising specific haplotypes of polymorphic variants predispose to isolated Hirschsprung disease. J. Med. Genet. 37, 572–578 (2000).
Lane, P.W. Association of megacolon with two recessive spotting genes in the mouse. J. Hered. 57, 181–183 (1966).
Shin, M.K., Russell, L.B. & Tilghman, S.M. Molecular characterization of four induced alleles at the Ednrb locus. Proc. Natl Acad. Sci. USA 94, 13105–13110 (1997).
Schuchardt, A., D'Agati, V., Larsson-Blomberg, L., Costantini, F. & Pachnis, V. Defects in the kidney and enteric nervous system of mice lacking the tyrosine kinase receptor Ret. Nature 367, 380–383 (1994).
Webster, W. Aganglionic megacolon in piebald-lethal mice. Arch. Pathol. 97, 111–117 (1974).
Hosoda, K. et al. Targeted and natural (piebald-lethal) mutations of endothelin-B receptor gene produce megacolon associated with spotted coat color in mice. Cell 79, 1267–1276 (1994).
Pachnis, V., Mankoo, B. & Costantini, F. Expression of the c-ret proto-oncogene during mouse embryogenesis. Development 119, 1005–1017 (1993).
Nataf, V., Lecoin, L., Eichmann, A. & Le Douarin, N.M. Endothelin-B receptor is expressed by neural crest cells in the avian embryo. Proc. Natl Acad. Sci. USA 93, 9645–9650 (1996).
Leibl, M.A. et al. Expression of endothelin 3 by mesenchymal cells of embryonic mouse caecum. Gut 44, 246–252 (1999).
Bourne, H.R. Signal transduction. Team blue sees red. Nature 376, 727–729 (1995).
van Biesen, T. et al. Receptor-tyrosine-kinase- and G β γ-mediated MAP kinase activation by a common signalling pathway. Nature 376, 781–784 (1995).
Prenzel, N. et al. EGF receptor transactivation by G-protein-coupled receptors requires metalloproteinase cleavage of proHB-EGF. Nature 402, 884–888 (1999).
Luttrell, L.M., Daaka, Y. & Lefkowitz, R.J. Regulation of tyrosine kinase cascades by G-protein-coupled receptors. Curr. Opin. Cell. Biol. 11, 177–183 (1999).
Auricchio, A. et al. Double heterozygosity for a RET substitution interfering with splicing and an EDNRB missense mutation in Hirschsprung disease. Am. J. Hum. Genet. 64, 1216–1221 (1999).
Rhim, H. et al. Spatially restricted hypopigmentation associated with an Ednrbs-modifying locus on mouse chromosome 10. Genome Res. 10, 17–29 (2000).
Miller, S.A., Dykes, D.D. & Polesky, H.F. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 16, 1215 (1988).
Weissenbach, J. et al. A second-generation linkage map of the human genome. Nature 359, 794–801 (1992).
Ceccherini, I. et al. Identification of the Cys634→Tyr mutation of the RET proto-oncogene in a pedigree with multiple endocrine neoplasia type 2A and localized cutaneous lichen amyloidosis. J. Endocrinol. Invest. 17, 201–204 (1994).
Mulligan, L.M. et al. Germ-line mutations of the RET proto-oncogene in multiple endocrine neoplasia type 2A. Nature 363, 458–460 (1993).
Lairmore, T.C. et al. A 1.5-megabase yeast artificial chromosome contig from human chromosome 10q11.2 connecting three genetic loci (RET, D10S94, and D10S102) closely linked to the MEN2A locus. Proc. Natl Acad. Sci. USA 90, 492–496 (1993).
Kent, W.J. BLAT—the BLAST-like alignment tool. Genome Res. 12, 656–664 (2002).
Gordon, D., Abajian, C. & Green, P. Consed: a graphical tool for sequence finishing. Genome Res. 8, 195–202 (1998).
Andrew, S.D., Delhanty, P.J., Mulligan, L.M. & Robinson, B.G. Sp1 and Sp3 transactivate the RET proto-oncogene promoter. Gene 256, 283–291 (2000).
Acknowledgements
We thank previous and current members of the Chakravarti lab for their contributions to this study, helpful discussions and comments on this manuscript; J. Scott for invaluable help in family recruitment; and A. Lynn for map construction. This study was begun at the Department of Genetics of Case Western Reserve University and is a portion of the Ph.D. dissertation of M.M.C.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
A.C. is a paid member of the Scientific Advisory Board of Affymetrix. The terms of this arrangement are being managed by Johns Hopkins University in accordance with its conflict of interest policies.
Rights and permissions
About this article
Cite this article
Carrasquillo, M., McCallion, A., Puffenberger, E. et al. Genome-wide association study and mouse model identify interaction between RET and EDNRB pathways in Hirschsprung disease. Nat Genet 32, 237–244 (2002). https://doi.org/10.1038/ng998
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ng998
This article is cited by
-
Stem cell-based therapy for hirschsprung disease, do we have the guts to treat?
Gene Therapy (2022)
-
Copy number variations in candidate genomic regions confirm genetic heterogeneity and parental bias in Hirschsprung disease
Orphanet Journal of Rare Diseases (2019)
-
Hirschsprung disease — integrating basic science and clinical medicine to improve outcomes
Nature Reviews Gastroenterology & Hepatology (2018)
-
Advances in understanding the association between Down syndrome and Hirschsprung disease (DS–HSCR)
Pediatric Surgery International (2018)
-
Hirschsprung’s disease: clinical dysmorphology, genes, micro-RNAs, and future perspectives
Pediatric Research (2017)