title: WNT3 involvement in human bladder exstrophy and cloaca development in zebrafish.
abstract: Bladder exstrophy, a severe congenital urological malformation when a child is born with an open urinary bladder, is the most common form of bladder exstrophy-epispadias complex (BEEC) with an incidence of 1:30,000 children of Caucasian descent. Recent studies suggest that WNT genes may contribute to the etiology of bladder exstrophy. Here, we evaluated WNT-pathway genes in 20 bladder exstrophy patients using massively parallel sequencing. In total 13 variants were identified in WNT3, WNT6, WNT7A, WNT8B, WNT10A, WNT11, WNT16, FZD5, LRP1 and LRP10 genes and predicted as potentially disease causing, of which seven variants were novel. One variant, identified in a patient with a de novo nonsynonymous substitution in WNT3 (p.Cys91Arg), was further evaluated in zebrafish. Knock down of wnt3 in zebrafish showed cloaca malformations, including disorganization of the cloaca epithelium and expansion of the cloaca lumen. Our study suggests that the function of the WNT3 p.Cys91Arg variant was altered, since RNA overexpression of mutant Wnt3 RNA does not result in embryonic lethality as seen with wild-type WNT3 mRNA. Finally, we also mutation screened the WNT3 gene further in 410 DNA samples from BEEC cases and identified one additional mutation c.638G>A (p.Gly213Asp), which was paternally inherited. In aggregate our data support the involvement of WNT-pathway genes in BEEC and suggest that WNT3 in itself is a rare cause of BEEC.
authors.affiliation: Department of Women's and Children's Health and Center for Molecular Medicine, Department of Clinical Genetics and.
authors.forename: Izabella
authors.initials: I
authors.lastname: Baranowska Körberg
authors.affiliation: Department of Molecular Medicine and Surgery and Center of Molecular Medicine and.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hofmeister
authors.affiliation: Department of Women's and Children's Health and Center for Molecular Medicine.
authors.forename: Ellen
authors.initials: E
authors.lastname: Markljung
authors.affiliation: Department of Women's and Children's Health and Center for Molecular Medicine.
authors.forename: Jia
authors.initials: J
authors.lastname: Cao
authors.affiliation: Department of Molecular Medicine and Surgery and Center of Molecular Medicine and Department of Clinical Genetics and Science for Life Laboratory, Karolinska Institutet Science Park, SE-17165 Stockholm, Sweden.
authors.forename: Daniel
authors.initials: D
authors.lastname: Nilsson
authors.affiliation: Department of Clinical Chemistry and Clinical Pharmacology.
authors.forename: Michael
authors.initials: M
authors.lastname: Ludwig
authors.affiliation: Institute of Human Genetics, Department of Genomics, Life & Brain Centre and.
authors.forename: Markus
authors.initials: M
authors.lastname: Draaken
authors.affiliation: Department of Pediatric Surgery, Queen Silvias Children's Hospital, SE-41685 Gothenburg, Sweden.
authors.forename: Gundela
authors.initials: G
authors.lastname: Holmdahl
authors.affiliation: Department of Women's and Children's Health, Uppsala Academic Children Hospital, SE-75185 Uppsala, Sweden and.
authors.forename: Gillian
authors.initials: G
authors.lastname: Barker
authors.affiliation: Institute of Human Genetics, Department of Neonatology, Children's Hospital University of Bonn, D-53127 Bonn, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Reutter
authors.affiliation: Department of Clinical Neuroscience, Center for Molecular Medicine, Karolinska Institutet, SE-17176 Stockholm, Sweden.
authors.forename: Vladana
authors.initials: V
authors.lastname: Vukojević
authors.affiliation: Department of Pediatric Surgery, University Hospital Lund, SE-22185 Lund, Sweden.
authors.forename: Christina
authors.initials: C
authors.lastname: Clementson Kockum
authors.affiliation: Department of Women's and Children's Health and Center for Molecular Medicine, Department of Clinical Genetics and.
authors.forename: Johanna
authors.initials: J
authors.lastname: Lundin
authors.affiliation: Department of Molecular Medicine and Surgery and Center of Molecular Medicine and Department of Clinical Genetics and.
authors.forename: Anna
authors.initials: A
authors.lastname: Lindstrand
authors.affiliation: Department of Women's and Children's Health and Center for Molecular Medicine, Pediatric Surgery, Astrid Lindgren Children Hospital, Karolinska University Hospital, SE-17176 Stockholm, Sweden, agneta.nordenskjold@ki.se.
authors.forename: Agneta
authors.initials: A
authors.lastname: Nordenskjöld
mesh_terms: D000818:Animals; D001746:Bladder Exstrophy / Q000235:genetics*; D002988:Cloaca / Q000196:embryology* / Q000378:metabolism*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D056726:Genetic Association Studies; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D009154:Mutation; D041681:NIH 3T3 Cells; D016366:Open Reading Frames; D019683:Penetrance; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D011487:Protein Conformation; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics; D060508:Wnt3 Protein / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D060508:Wnt3 Protein