title: Mutational Spectrum of CYP24A1 Gene in a Cohort of Italian Patients with Idiopathic Infantile Hypercalcemia.
abstract: OBJECTIVE
  Loss-of-function mutations in the CYP24A1 gene, which encodes the vitamin D-24 hydroxylase, have been recognized as a cause of elevated 1,25-dihydroxyvitamin D concentrations, hypercalcemia, hypercalciuria, nephrocalcinosis and nephrolithiasis in infants and adults. As only a case report describing 2 adult patients has been reported in Italian population, we report here the mutation analysis of CYP24A1 gene in an Italian cohort of 12 pediatric and adult patients with idiopathic infantile hypercalcemia (IIH).
  
  
  METHODS
  We performed mutational screening of CYP24A1 gene in a cohort of 12 Italian patients: 8 children with nephrocalcinosis, hypercalcemia and PTH levels <10 pg/ml and 4 adult patients with nephrolithiasis, mild hypercalcemia and PTH levels <10 pg/ml from 11 unrelated Italian families. Clinical and biochemical data were collected. Genomic DNA was extracted from peripheral blood leucocytes using standard methods, and whole coding sequence of CYP24A1 gene was analysed in all patients and family members by polymerase chain reaction and direct sequencing. The potential pathogenicity of the newly identified missense mutations was evaluated by 3 different in silico approaches (Sorting Intolerant from Tolerant, Polyphen and Mutation Taster) and by comparative analysis in 14 different species using ClustalW software.
  
  
  RESULTS
  CYP24A1 bi-allelic mutations were found in 8 individuals from 7 Italian families (7/11; 64%). Overall, 6 different CYP24A1 mutations, including one small deletion (p.Glu143del), 4 missense mutations (p.Leu148Pro; p.Arg396Trp; p.Pro503Leu; p.Glu383Gln) and one nonsense mutation (p.Tyr220*) were identified. Two out of 6 mutations (p.Tyr220* and p.Pro503Leu) were not previously described. Moreover, a new CYP24A1 variant was identified by genetic screening of asymptomatic controls.
  
  
  CONCLUSIONS
  To the best of our knowledge, this is the first report of a CYP24A1 molecular analysis performed in an Italian cohort of adult and pediatric Italian patients. This study (1) confirms that CYP24A1 plays a causal role in some but not all cases of IIH (64%); (2) expands the spectrum of known CYP24A1 pathogenic mutations; (3) describes 2 hotspots detected in 50% of all Italian cases; and (4) emphasizes the importance of recognition and genetic diagnosis of CYP24A1 defects in infantile as well as adult hypercalcemia.
authors.affiliation: Nephrology, Dialysis and Transplantation Unit, Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Maddalena
authors.initials: M
authors.lastname: Gigante
authors.forename: Luisa
authors.initials: L
authors.lastname: Santangelo
authors.forename: Sterpeta
authors.initials: S
authors.lastname: Diella
authors.forename: Gianluca
authors.initials: G
authors.lastname: Caridi
authors.forename: Lucia
authors.initials: L
authors.lastname: Argentiero
authors.forename: Maria Michela
authors.initials: MM
authors.lastname: D''Alessandro
authors.forename: Marida
authors.initials: M
authors.lastname: Martino
authors.forename: Emma Diletta
authors.initials: ED
authors.lastname: Stea
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Ardissino
authors.forename: Vincenza
authors.initials: V
authors.lastname: Carbone
authors.forename: Silvana
authors.initials: S
authors.lastname: Pepe
authors.forename: Domenico
authors.initials: D
authors.lastname: Scrutinio
authors.forename: Silvio
authors.initials: S
authors.lastname: Maringhini
authors.forename: Gian Marco
authors.initials: GM
authors.lastname: Ghiggeri
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Grandaliano
authors.forename: Mario
authors.initials: M
authors.lastname: Giordano
authors.forename: Loreto
authors.initials: L
authors.lastname: Gesualdo
mesh_terms: D000328:Adult; D002648:Child; D002675:Child, Preschool; D015331:Cohort Studies; D005260:Female; D006801:Humans; D006934:Hypercalcemia / Q000235:genetics*; D007558:Italy; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010375:Pedigree; D065668:Vitamin D3 24-Hydroxylase / Q000235:genetics*; D055815:Young Adult
chemical_list: C585549:CYP24A1 protein, human; D065668:Vitamin D3 24-Hydroxylase