title: Molecular basis of non-syndromic hypospadias: systematic mutation screening and genome-wide copy-number analysis of 62 patients.
abstract: OBJECTIVE
  What percentage of cases with non-syndromic hypospadias can be ascribed to mutations in known causative/candidate/susceptibility genes or submicroscopic copy-number variations (CNVs) in the genome?
  
  
  CONCLUSIONS
  Monogenic and digenic mutations in known causative genes and cryptic CNVs account for >10% of cases with non-syndromic hypospadias. While known susceptibility polymorphisms appear to play a minor role in the development of this condition, further studies are required to validate this observation.
  
  
  BACKGROUND
  Fifteen causative, three candidate, and 14 susceptible genes, and a few submicroscopic CNVs have been implicated in non-syndromic hypospadias.
  
  
  METHODS
  Systematic mutation screening and genome-wide copy-number analysis of 62 patients.
  
  
  METHODS
  The study group consisted of 57 Japanese and five Vietnamese patients with non-syndromic hypospadias. Systematic mutation screening was performed for 25 known causative/candidate/susceptibility genes using a next-generation sequencer. Functional consequences of nucleotide alterations were assessed by in silico assays. The frequencies of polymorphisms in the patient group were compared with those in the male general population. CNVs were analyzed by array-based comparative genomic hybridization and characterized by fluorescence in situ hybridization.
  
  
  RESULTS
  Seven of 62 patients with anterior or posterior hypospadias carried putative pathogenic mutations, such as hemizygous mutations in AR, a heterozygous mutation in BNC2, and homozygous mutations in SRD5A2 and HSD3B2. Two of the seven patients had mutations in multiple genes. We did not find any rare polymorphisms that were abundant specifically in the patient group. One patient carried mosaic dicentric Y chromosome.
  
  
  CONCLUSIONS
  The patient group consisted solely of Japanese and Vietnamese individuals and clinical and hormonal information of the patients remained rather fragmentary. In addition, mutation analysis focused on protein-altering substitutions.
  
  
  CONCLUSIONS
  Our data provide evidence that pathogenic mutations can underlie both mild and severe hypospadias and that HSD3B2 mutations cause non-syndromic hypospadias as a sole clinical manifestation. Most importantly, this is the first report documenting possible oligogenicity of non-syndromic hypospadias.
  
  
  BACKGROUND
  This study was funded by the Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology; by the Grant-in-Aid from the Japan Society for the Promotion of Science; by the Grants from the Ministry of Health, Labour and Welfare, from the National Center for Child Health and Development and from the Takeda Foundation. The authors have no competing interests to disclose.
  
  
  BACKGROUND
  Not applicable.
authors.affiliation: Department of Molecular Endocrinology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan Department of Renal and Genitourinary Surgery, Hokkaido University Graduate School of Medicine, Sapporo 060-8638, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kon
authors.affiliation: Department of Molecular Endocrinology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Suzuki
authors.affiliation: Department of Endocrinology, The Vietnam National Hospital of Pediatrics, Hanoi, Vietnam.
authors.forename: V C
authors.initials: VC
authors.lastname: Dung
authors.affiliation: Department of Endocrinology and Metabolism, Tokyo Metropolitan Children's Medical Center, Tokyo 183-8561, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Hasegawa
authors.affiliation: Department of Renal and Genitourinary Surgery, Hokkaido University Graduate School of Medicine, Sapporo 060-8638, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Mitsui
authors.affiliation: Department of Endocrinology and Metabolism, Kanagawa Children's Medical Center, Yokohama 232-0066, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Muroya
authors.affiliation: Division of Urology, National Center for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ueoka
authors.affiliation: Department of Pediatrics, Toyama Prefectural Central Hospital, Toyama 930-0975, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Igarashi
authors.affiliation: Department of Pediatrics, Niigata University School of Medicine, Niigata 951-8510, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Nagasaki
authors.affiliation: Department of Pediatrics, Dokkyo Medical University Koshigaya Hospital, Koshigaya 343-0845, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Oto
authors.affiliation: Division of Endocrinology and Metabolism, Aichi Children's Health and Medical Center, Obu 474-8710, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Hamajima
authors.affiliation: Department of Urology, Aichi Children's Health and Medical Center, Obu 474-8710, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yoshino
authors.affiliation: Department of Molecular Endocrinology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Igarashi
authors.affiliation: Department of Molecular Endocrinology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kato-Fukui
authors.affiliation: Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Nakabayashi
authors.affiliation: Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Hayashi
authors.affiliation: Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Hata
authors.affiliation: National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Matsubara
authors.affiliation: Department of Renal and Genitourinary Surgery, Hokkaido University Graduate School of Medicine, Sapporo 060-8638, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Moriya
authors.affiliation: Department of Molecular Endocrinology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan Department of Pediatrics, Hamamatsu University School of Medicine, Hamamatsu 431-3192, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Ogata
authors.affiliation: Department of Renal and Genitourinary Surgery, Hokkaido University Graduate School of Medicine, Sapporo 060-8638, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Nonomura
authors.affiliation: Department of Molecular Endocrinology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan fukami-m@ncchd.go.jp.
authors.forename: M
authors.initials: M
authors.lastname: Fukami
mesh_terms: D056915:DNA Copy Number Variations; D004252:DNA Mutational Analysis; D020022:Genetic Predisposition to Disease; D006801:Humans; D007021:Hypospadias / Q000235:genetics*; D008297:Male; D011110:Polymorphism, Genetic
keywords: copy-number; hypospadias; mutation; polymorphism; susceptibility