File Download
 
 
Supplementary

Conference Paper: Genetic study of a family segregating Shah-Waardenburg syndrome
  • Basic View
  • Metadata View
  • XML View
TitleGenetic study of a family segregating Shah-Waardenburg syndrome
 
AuthorsCui, L
Wong, EHM
Zhu, J
Firmato de, AM
Tam, PKH
Garcia-Barcelo, MM
 
Issue Date2012
 
CitationThe 3rd International Symposium on Development of the Enteric Nervous System: Cells, Signals and Genes, Hong Kong, 25-28 March 2012 [How to Cite?]
 
AbstractBackground: Type IV Waardenburg syndrome (WS4, MIM_277580), also known as Shah-Waardenburg syndrome or Waardenburg-Hirschsprung disease, is a congenital developmental disorder characterized by pigmentary abnormalities of the skin, eyes and hair, sensorineural deafness and Hirschsprung disease. The WS4 is caused by mutations in any of the following three genes: endothelin-3(EDN3), endothelin-B receptor (EDNRB), or SOX10. Materials and methods: Exons and exon/intron boundaries of the three candidate genes (EDN3, EDNRB, SOX10) were screened for mutations by direct sequencing of PCR products in a three-generation family (14 individuals) whose members were affected with WS4 or only HSCR. The family members were also genotyped for 2.5 million genetic markers (single nucleotide polymorphisms -SNP- and copy number variation probes -CNVs-) using Illumina Human Omni2.5-quad BeadChip. After quality control and pruning of SNPs in high linkage-disequilibrium, the Merlin software was used for parametric and non-parametric linkage. To test the effect of the mutation identified, wild-type and mutated mammalian expression vectors encoding for EDNRB isoforms (with green fluorescent protein -GFP-) were/will (isoform 3) transiently transfected in Human Embryonic Kidney 293 cells. Results: Direct sequencing revealed a heterozygous novel nonsense mutation in EDNRB (Chromosome 13). This mutation corresponds to M1V of the EDNRB isoforms 1 and 2 and M91V of the EDNRB isoform 3. It was present in four affected and two unaffected family members. The PolyPhen and SIFT prediction for M1V (isoforms 1&2) and for M91V (isoform 3) was damaging (abolishes the ATG start site) and benign respectively. Importantly, the 3 EDNRB transcripts are expressed in the human new-born gut. HEK 293 cells transfected with vector with wild-type EDNRB-GFP showed mainly a cell membrane localization, while the M1V mutated EDNRB-GFP revealed a clustered localization in cytoplasm indicating that EDNRB receptor cannot translocate to cell membrane. Analysis of M91V is underway. Our linkage analysis indicated that additional genes may be contributing to the phenotype. Conclusion: We found and characterized a novel nonsense mutation. The EDNRB mutation is deleterious in isoforms 1 and 2 (M1V) but predicted benign in isoform 3 (M91V) (in vitro is study underway). The variable manifestation of the disease may depend on the ratio between the EDNRB isoforms expressed in gut or/and additional genetic factors involved in the pathogenesis of the disease.
 
DescriptionSession: Genetics of Hirschsprung’s Disease, no. E28
Poster presentation
 
DC FieldValue
dc.contributor.authorCui, L
 
dc.contributor.authorWong, EHM
 
dc.contributor.authorZhu, J
 
dc.contributor.authorFirmato de, AM
 
dc.contributor.authorTam, PKH
 
dc.contributor.authorGarcia-Barcelo, MM
 
dc.date.accessioned2012-09-20T08:20:33Z
 
dc.date.available2012-09-20T08:20:33Z
 
dc.date.issued2012
 
dc.description.abstractBackground: Type IV Waardenburg syndrome (WS4, MIM_277580), also known as Shah-Waardenburg syndrome or Waardenburg-Hirschsprung disease, is a congenital developmental disorder characterized by pigmentary abnormalities of the skin, eyes and hair, sensorineural deafness and Hirschsprung disease. The WS4 is caused by mutations in any of the following three genes: endothelin-3(EDN3), endothelin-B receptor (EDNRB), or SOX10. Materials and methods: Exons and exon/intron boundaries of the three candidate genes (EDN3, EDNRB, SOX10) were screened for mutations by direct sequencing of PCR products in a three-generation family (14 individuals) whose members were affected with WS4 or only HSCR. The family members were also genotyped for 2.5 million genetic markers (single nucleotide polymorphisms -SNP- and copy number variation probes -CNVs-) using Illumina Human Omni2.5-quad BeadChip. After quality control and pruning of SNPs in high linkage-disequilibrium, the Merlin software was used for parametric and non-parametric linkage. To test the effect of the mutation identified, wild-type and mutated mammalian expression vectors encoding for EDNRB isoforms (with green fluorescent protein -GFP-) were/will (isoform 3) transiently transfected in Human Embryonic Kidney 293 cells. Results: Direct sequencing revealed a heterozygous novel nonsense mutation in EDNRB (Chromosome 13). This mutation corresponds to M1V of the EDNRB isoforms 1 and 2 and M91V of the EDNRB isoform 3. It was present in four affected and two unaffected family members. The PolyPhen and SIFT prediction for M1V (isoforms 1&2) and for M91V (isoform 3) was damaging (abolishes the ATG start site) and benign respectively. Importantly, the 3 EDNRB transcripts are expressed in the human new-born gut. HEK 293 cells transfected with vector with wild-type EDNRB-GFP showed mainly a cell membrane localization, while the M1V mutated EDNRB-GFP revealed a clustered localization in cytoplasm indicating that EDNRB receptor cannot translocate to cell membrane. Analysis of M91V is underway. Our linkage analysis indicated that additional genes may be contributing to the phenotype. Conclusion: We found and characterized a novel nonsense mutation. The EDNRB mutation is deleterious in isoforms 1 and 2 (M1V) but predicted benign in isoform 3 (M91V) (in vitro is study underway). The variable manifestation of the disease may depend on the ratio between the EDNRB isoforms expressed in gut or/and additional genetic factors involved in the pathogenesis of the disease.
 
dc.descriptionSession: Genetics of Hirschsprung’s Disease, no. E28
 
dc.descriptionPoster presentation
 
dc.identifier.citationThe 3rd International Symposium on Development of the Enteric Nervous System: Cells, Signals and Genes, Hong Kong, 25-28 March 2012 [How to Cite?]
 
dc.identifier.hkuros211258
 
dc.identifier.urihttp://hdl.handle.net/10722/165593
 
dc.languageeng
 
dc.relation.ispartofInternational Symposium on Development of the Enteric Nervous System: Cells, Signals and Genes
 
dc.titleGenetic study of a family segregating Shah-Waardenburg syndrome
 
dc.typeConference_Paper
 
<?xml encoding="utf-8" version="1.0"?>
<item><contributor.author>Cui, L</contributor.author>
<contributor.author>Wong, EHM</contributor.author>
<contributor.author>Zhu, J</contributor.author>
<contributor.author>Firmato de, AM</contributor.author>
<contributor.author>Tam, PKH</contributor.author>
<contributor.author>Garcia-Barcelo, MM</contributor.author>
<date.accessioned>2012-09-20T08:20:33Z</date.accessioned>
<date.available>2012-09-20T08:20:33Z</date.available>
<date.issued>2012</date.issued>
<identifier.citation>The 3rd International Symposium on Development of the Enteric Nervous System: Cells, Signals and Genes, Hong Kong, 25-28 March 2012</identifier.citation>
<identifier.uri>http://hdl.handle.net/10722/165593</identifier.uri>
<description>Session: Genetics of Hirschsprung&#8217;s Disease, no. E28</description>
<description>Poster presentation</description>
<description.abstract>Background: Type IV Waardenburg syndrome 
(WS4, MIM_277580), also known as 
Shah-Waardenburg syndrome or 
Waardenburg-Hirschsprung disease, is a congenital 
developmental disorder characterized by 
pigmentary abnormalities of the skin, eyes and hair, 
sensorineural deafness and Hirschsprung disease. 
The WS4 is caused by mutations in any of the 
following three genes: endothelin-3(EDN3), 
endothelin-B receptor (EDNRB), or SOX10. 

Materials and methods: Exons and exon/intron 
boundaries of the three candidate genes (EDN3, 
EDNRB, SOX10) were screened for mutations by 
direct sequencing of PCR products in a 
three-generation family (14 individuals) whose 
members were affected with WS4 or only HSCR. 
The family members were also genotyped for 2.5 
million genetic markers (single nucleotide 
polymorphisms -SNP- and copy number variation 
probes -CNVs-) using Illumina Human 
Omni2.5-quad BeadChip. After quality control and 
pruning of SNPs in high linkage-disequilibrium, 
the Merlin software was used for parametric and 
non-parametric linkage. To test the effect of the 
mutation identified, wild-type and mutated 
mammalian expression vectors encoding for 
EDNRB isoforms (with green fluorescent protein 
-GFP-) were/will (isoform 3) transiently 
transfected in Human Embryonic Kidney 293 cells.


Results: Direct sequencing revealed a 
heterozygous novel nonsense mutation in EDNRB 
(Chromosome 13). This mutation corresponds to 
M1V of the EDNRB isoforms 1 and 2 and M91V of 
the EDNRB isoform 3. It was present in four 
affected and two unaffected family members. The 
PolyPhen and SIFT prediction for M1V (isoforms 
1&amp;2) and for M91V (isoform 3) was damaging 
(abolishes the ATG start site) and benign 
respectively. Importantly, the 3 EDNRB transcripts 
are expressed in the human new-born gut. HEK 293 
cells transfected with vector with wild-type EDNRB-GFP showed mainly a cell membrane 
localization, while the M1V mutated EDNRB-GFP 
revealed a clustered localization in cytoplasm 
indicating that EDNRB receptor cannot translocate 
to cell membrane. Analysis of M91V is underway. 
Our linkage analysis indicated that additional genes 
may be contributing to the phenotype. 
 
Conclusion: We found and characterized a novel 
nonsense mutation. The EDNRB mutation is 
deleterious in isoforms 1 and 2 (M1V) but 
predicted benign in isoform 3 (M91V) (in vitro is 
study underway). The variable manifestation of the 
disease may depend on the ratio between the 
EDNRB isoforms expressed in gut or/and 
additional genetic factors involved in the 
pathogenesis of the disease.</description.abstract>
<language>eng</language>
<relation.ispartof>International Symposium on Development of the Enteric Nervous System: Cells, Signals and Genes</relation.ispartof>
<title>Genetic study of a family segregating Shah-Waardenburg syndrome</title>
<type>Conference_Paper</type>
<identifier.hkuros>211258</identifier.hkuros>
</item>