title: CBX2 gene analysis in patients with 46,XY and 46,XX gonadal disorders of sex development.
abstract: OBJECTIVE
  To investigate a cohort of patients with gonadal disorders of sex development (DSD) for causative CBX2 gene mutations and or gene copy number changes.
  
  
  METHODS
  Genetic association study.
  
  
  METHODS
  University laboratory and tertiary university-based referral center.
  
  
  METHODS
  47 patients with different forms of 46,XY or 46,XX gonadal DSD.
  
  
  METHODS
  CBX2 gene sequencing and development of a synthetic probe set for multiplex ligation probe amplification (MLPA) to detect CBX2 copy number changes, and reverse-transcriptase polymerase chain reaction (RT-PCR) to evaluate CBX2 expression in two different cell-line types.
  
  
  METHODS
  Gene sequence alteration and or partial or complete gene copy number variations, and detection of CBX2 mRNA isoforms.
  
  
  RESULTS
  We detected 10 sequence alterations, 9 reported single nucleotide polymorphisms (SNPs), and a previously unreported variant. This was a silent c.1356G>A transition that may represent a normal variant. A rare SNP (c.1411C>G, p.471Pro>Ala) was found in heterozygous form in one patient. No deletions or duplications were detected by MLPA. Expression of both CBX2 mRNA isoforms was documented in gonadal fibroblasts and Epstein Barr virus (EBV)-transformed lymphocytes.
  
  
  CONCLUSIONS
  No pathogenic CBX2 mutation was detected. Both CBX2 isoforms are expressed in gonadal fibroblasts and EBV-transformed lymphocytes. This study does not support CBX2 gene disruption as a common cause of gonadal DSD.
authors.affiliation: Department of Molecular Medicine and Surgery, Karolinska Institutet Stockholm, Karolinska University Hospital, Stockholm, Sweden. ameli.norling@ki.se
authors.forename: Ameli
authors.initials: A
authors.lastname: Norling
authors.forename: Angelica Lindén
authors.initials: AL
authors.lastname: Hirschberg
authors.forename: Erik
authors.initials: E
authors.lastname: Iwarsson
authors.forename: Anna
authors.initials: A
authors.lastname: Wedell
authors.forename: Michela
authors.initials: M
authors.lastname: Barbaro
mesh_terms: D058531:46, XX Testicular Disorders of Sex Development / Q000235:genetics*; D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D005260:Female; D005347:Fibroblasts / Q000502:physiology; D018628:Gene Dosage / Q000235:genetics; D006061:Gonadal Dysgenesis, 46,XY / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D017354:Point Mutation / Q000235:genetics; D063150:Polycomb Repressive Complex 1 / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D016649:Primary Ovarian Insufficiency / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055815:Young Adult
chemical_list: C570479:CBX2 protein, human; D063150:Polycomb Repressive Complex 1