title: Prevalence of mutations in TSHR, GNAS, PRKAR1A and RAS genes in a large series of toxic thyroid adenomas from Galicia, an iodine-deficient area in NW Spain.
abstract: OBJECTIVE
  Toxic thyroid adenoma (TA) is a common cause of hyperthyroidism. Mutations in the TSH receptor (TSHR) gene, and less frequently in the adenylate cyclase-stimulating G alpha protein (GNAS) gene, are well established causes of TA in Europe. However, genetic causes of TA remain unknown in a small percentage of cases. We report the first study to investigate mutations in TSHR, GNAS, protein kinase, cAMP-dependent, regulatory, type I alpha (PRKAR1A) and RAS genes, in a large series of TA from Galicia, an iodine-deficient region in NW Spain.
  
  
  METHODS
  Eighty-five TA samples were obtained surgically from 77 hyperthyroid patients, operated on for treatment of non-autoimmune toxic nodular goitre. After DNA extraction, all coding exons of TSHR, GNAS and PRKAR1A genes, and exons 2 and 3 of HRAS, KRAS and NRAS were amplified by PCR and sequenced. Previously unreported mutants were cloned in expression vectors and their basal constitutive activities were determined by quantification of cAMP response element (CRE)-luciferase activity in CO7 cells transfected with wild-type and mutant plasmids.
  
  
  RESULTS
  TSHR gene mutations were found in 52 (61.2%) samples, GNAS gene mutations in 4 (4.71%) samples and no PRKAR1A or RAS mutations were found. Only three previously unreported mutations were found, two affecting the TSHR, A623F and I635V, and one affecting the G-protein alpha-subunit (Gsalpha), L203P. All mutant proteins showed higher CRE-luciferase activity than their wild-type counterparts.
  
  
  CONCLUSIONS
  TA in a hyperthyroid population living in Galicia, a Spanish iodine-deficient region, harbours elevated frequencies of TSHR and GNAS mutations activating the cAMP pathway. However, the genetic cause of TA was undetermined in 34% of the TA samples.
authors.affiliation: Unidade de Enfermedades Tiroideas e Metabólicas (UETeM), Complexo Hospitalario Universitary de Santiago (CHUS), University of Santiago de Compostela, Santiago de Compostela, 15705, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Palos-Paz
authors.forename: O
authors.initials: O
authors.lastname: Perez-Guerra
authors.forename: J
authors.initials: J
authors.lastname: Cameselle-Teijeiro
authors.forename: C
authors.initials: C
authors.lastname: Rueda-Chimeno
authors.forename: F
authors.initials: F
authors.lastname: Barreiro-Morandeira
authors.forename: J
authors.initials: J
authors.lastname: Lado-Abeal
authors.forename: D
authors.initials: D
authors.lastname: Araujo Vilar
authors.forename: R
authors.initials: R
authors.lastname: Argueso
authors.forename: O
authors.initials: O
authors.lastname: Barca
authors.forename: M
authors.initials: M
authors.lastname: Botana
authors.forename: J M
authors.initials: JM
authors.lastname: Cabezas-Agrícola
authors.forename: P
authors.initials: P
authors.lastname: Catalina
authors.forename: L
authors.initials: L
authors.lastname: Dominguez Gerpe
authors.forename: T
authors.initials: T
authors.lastname: Fernandez
authors.forename: A
authors.initials: A
authors.lastname: Mato
authors.forename: A
authors.initials: A
authors.lastname: Nuño
authors.forename: M
authors.initials: M
authors.lastname: Penin
authors.forename: B
authors.initials: B
authors.lastname: Victoria
mesh_terms: D000236:Adenoma / Q000453:epidemiology / Q000235:genetics*; D000328:Adult; D000368:Aged; D002864:Chromogranins; D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit / Q000235:genetics*; D019353:Endemic Diseases; D005260:Female; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics*; D011905:Genes, ras / Q000235:genetics*; D020022:Genetic Predisposition to Disease / Q000453:epidemiology; D006801:Humans; D006980:Hyperthyroidism / Q000453:epidemiology / Q000235:genetics; D007455:Iodine / Q000172:deficiency; D008297:Male; D008875:Middle Aged; D009154:Mutation; D015995:Prevalence; D011989:Receptors, Thyrotropin / Q000235:genetics*; D013030:Spain; D013964:Thyroid Neoplasms / Q000453:epidemiology / Q000235:genetics*
chemical_list: D002864:Chromogranins; D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit; C517834:PRKAR1A protein, human; D011989:Receptors, Thyrotropin; D007455:Iodine; C406732:GNAS protein, human; D019205:GTP-Binding Protein alpha Subunits, Gs