A novel CYP21A2 mutation identified a patient with classical 21-hydroxylase deficiency

sem informação8627575XXVI Annual Meeting of the Latin American Pediatric Endocrinology Societ

Preserved Fertility In A Patient With Gynecomastia Associated With The P.pro695ser Mutation In The Androgen Receptor

The androgen insensitivity syndrome (AIS) is described as a dysfunction of the androgen receptor (AR) in 46,XY individuals, which can be associated with mutations in the AR gene or can be due to unknown mechanisms. Different mutations in AIS generally cause variable phenotypes that range from a complete hormone resistance to a mild form usually associated with male infertility. The purpose of this study was to search for mutations in the AR gene in a fertile man with gynecomastia and to evaluate the influence of the mutation on the AR transactivation ability. Sequencing of the AR gene revealed the p.Pro695Ser mutation. It is located within the AR ligand-binding domain. Bioinformatics analysis indicated a deleterious role, which was verified after testing transactivation activity and N-/C-terminal (N/C) interaction by in vitro expression of a reporter gene and 2-hybrid assays. p.Pro695Ser showed low levels of both transactivation activity and N/C interaction at low dihydrotestosterone (DHT) conditions. As the ligand concentration increased, both transactivation activity and N/C interaction also increased and reached normal levels. Therefore, this study provides functional insights for the p.Pro695Ser mutation described here for the first time in a patient with mild AIS. The expression profile of p.Pro695Ser not only correlates to the patient's phenotype, but also suggests that a high-dose DHT therapy may overcome the functional deficit of the mutant AR.86350355Brinkmann, A.O., Molecular basis of androgen insensitivity (2001) Mol Cell Endocrinol, 179, pp. 105-109Brinkmann, A.O., Molecular mechanisms of androgen action Androgen Action: Methods in Molecular Biology, pp. 3-24. , Saatcioglu F (ed): Humana Press, New York 2011)Chang, C.S., Kokontis, J., Liao, S.T., Structural analysis of complementary dna and amino acid sequences of human and rat androgen receptors (1988) Proc Natl Acad Sci USA, 85, pp. 7211-7215Cheikhelard, A., Morel, Y., Thibaud, E., Lortat-Jacob, S., Jaubert, F., Long-term followup and comparison between genotype and phenotype in 29 cases of complete androgen insensitivity syndrome (2008) J Urol, 180, pp. 1496-1501Chu, J., Zhang, R., Zhao, Z., Zou, W., Han, Y., Male fertility is compatible with an arg(840)cys substitution in the ar in a large chinese family affected with divergent phenotypes of ar insensitivity syndrome (2002) J Clin Endocrinol Metab, 87, pp. 347-351Clinckemalie, L., Vanderschueren, D., Boonen, S., Claessens, F., The hinge region in androgen receptor control (2012) Mol Cell Endocrinol, 358, pp. 1-8Dörk, T., Schnieders, F., Jakubiczka, S., Wieacker, P., Schroeder-Kurth, T., Schmidtke, J., A new missense substitution at a mutational hot spot of the androgen receptor in siblings with complete androgen insensitivity syndrome (1998) Hum Mutat, 11, pp. 337-339Estébanez-Perpiñá, E., Arnold, L.A., Nguyen, P., Rodrigues, E.D., Mar, E., A surface on the androgen receptor that allosterically regulates coactivator binding (2007) Proc Natl Acad Sci USA, 104, pp. 16074-16079Giwercman, A., Kledal, T., Schwartz, M., Giwercman, Y.L., Leffers, H., Preserved male fertility despite decreased androgen sensitivity caused by a mutation in the ligand-binding domain of the androgen receptor gene (2000) J Clin Endocrinol Metab, 85, pp. 2253-2259He, B., Wilson, E.M., The nh(2)-terminal and carboxyl-terminal interaction in the human androgen receptor (2002) Mol Genet Metab, 75, pp. 293-298He, B., Gampe, R.T., Jr., Hnat, A.T., Faggart, J.L., Minges, J.T., Probing the functional link between androgen receptor coactivator and ligandbinding sites in prostate cancer and androgen insensitivity (2006) J Biol Chem, 281, pp. 6648-6663Helsen, C., Kerkhofs, S., Clinckemalie, L., Spans, L., Laurent, M., Structural basis for nuclear hormone receptor dna binding (2012) Mol Cell Endocrinol, 348, pp. 411-417Hiort, O., Sinnecker, G.H., Holterhus, P.M., Nitsche, E.M., Kruse, K., Inherited and de novo androgen receptor gene mutations: Investigation of single-case families (1998) J Pediat, 132, pp. 939-943Hiort, O., Holterhus, P.M., Horter, T., Schulze, W., Kremke, B., Significance of mutations in the androgen receptor gene in males with idiopathic infertility (2000) J Clin Endocrinol Metab, 85, pp. 2810-2815Hughes, I.A., Disorders of sex development: A new definition and classification (2008) Best Pract Res Clin Endocrinol Metab, 22, pp. 119-134Hughes, I.A., Werner, R., Bunch, T., Hiort, O., Androgen insensitivity syndrome (2012) Semin Reprod Med, 30, pp. 432-442Jääskeläinen, J., Deeb, A., Schwabe, J.W., Mongan, N.P., Martin, H., Hughes, I.A., Human androgen receptor gene ligand-binding-domain mutations leading to disrupted interaction between the n-and c-terminal domains (2006) J Mol Endocrinol, 36, pp. 361-368Larrea, F., Benavides, G., Scaglia, H., Kofman-Alfaro, S., Ferrusca, E., Gynecomastia as a familial incomplete male pseudohermaphroditism type 1: A limited androgen resistance syndrome (1978) J Clin Endocrinol Metab, 46, pp. 961-970Lubahn, D.B., Joseph, D.R., Sullivan, P.M., Willard, H.F., French, F.S., Wilson, E.M., Cloning of human androgen receptor complementary DNA and localization to the X chromosome (1988) Science, 240, pp. 327-330Maclean, H.E., Warne, G.L., Zajac, J.D., Localization of functional domains in the androgen receptor (1997) J Steroid Biochem Mol Biol, 62, pp. 233-242Morris, J.M., The syndrome of testicular feminization in male pseudohermaphroditism (1953) Am J Obstet Gynecol, 65, pp. 1192-1211Petroli, R.J., Maciel-Guerra, A.T., Soardi, F.C., De Calais, F.L., Guerra-Junior, G., De Mello, M.P., Severe forms of partial androgen insensitivity syndrome due to p.l830f novel mutation in androgen receptor gene in a brazilian family (2011) BMC Res Notes, 4, p. 173Powell, S.M., Christiaens, V., Voulgaraki, D., Waxman, J., Claessens, F., Bevan, C.L., Mechanisms of androgen receptor signalling via steroid receptor coactivator-1 in prostate (2004) Endocr Relat Cancer, 11, pp. 117-130Quigley, C.A., De Bellis, A., Marschke, K.B., El-Awady, M.K., Wilson, E.M., French, F.S., Androgen receptor defects: Historical, clinical, and molecular perspectives (1995) Endocr Rev, 16, pp. 271-321Sack, J.S., Kish, K.F., Wang, C., Attar, R.M., Kiefer, S.E., Crystallographic structures of the ligandbinding domains of the androgen receptor and its t877a mutant complexed with the natural agonist dihydrotestosterone (2001) Proc Natl Acad Sci USA, 98, pp. 4904-4909Trapman, J., Klaassen, P., Kuiper, G.G., Van Der Korput, J.A., Faber, P.W., Cloning, structure and expression of a cdna encoding the human androgen receptor (1988) Biochem Biophys Res Commun, 153, pp. 241-248Werner, R., Schütt, J., Hannema, S., Röpke, A., Wieacker, P., Androgen receptor gene mutations in androgen insensitivity syndrome cause distinct patterns of reduced activation of androgen-responsive promoter constructs (2006) J Steroid Biochem Mol Biol, 101, pp. 1-10Werner, R., Zhan, J., Gesing, J., Struve, D., Hiort, O., In-vitro characterization of androgen receptor mutations associated with complete androgen insensitivity syndrome reveals distinct functional deficits (2008) Sex Dev, 2, pp. 73-83Zhou, J., Liu, B., Geng, G., Wu, J.H., Study of the impact of the t877a mutation on ligand-induced helix-12 positioning of the androgen receptor resulted in design and synthesis of novel antiandrogens (2010) Proteins, 78, pp. 623-63

Two Novel Mutations In The Thyroid Hormone Receptor β In Patients With Resistance To Thyroid Hormone (rth β): Clinical, Biochemical, And Molecular Data

The syndrome of resistance to thyroid hormone (RTH β) is an inherited disorder characterized by variable tissue hyposensitivity to 3,5,30-l-triiodothyronine (T3), with persistent elevation of free-circulating T3 (FT3) and free thyroxine (FT4) levels in association with nonsuppressed serum thyrotropin (TSH). Clinical presentation is variable and the molecular analysis of THRB gene provides a short cut diagnosis. Here, we describe 2 cases in which RTH β was suspected on the basis of laboratory findings. The diagnosis was confirmed by direct THRB sequencing that revealed 2 novel mutations: the heterozygous p.Ala317Ser in subject 1 and the heterozygous p.Arg438Pro in subject 2. Both mutations were shown to be deleterious by SIFT, PolyPhen, and Align GV-GD predictive methods

A case of 46,XY DSD due to a novel mutation in the HSD17B3 gene

sem informação8626161XXVI Annual Meeting of the Latin American Pediatric Endocrinology Societ

46,xx Dsd And Antley-bixler Syndrome Due To Novel Mutations In The Cytochrome P450 Oxidoreductase Gene [dds 46,xx E Síndrome De Antley-bixler Causada Por Novas Mutações No Gene Da Enzima P450 Oxidorredutase]

Deficiency of the enzyme P450 oxidoreductase is a rare form of congenital adrenal hyperplasia with characteristics of combined and partial impairments in steroidogenic enzyme activities, as P450 oxidoreductase transfers electrons to CYP21A2, CYP17A1, and CYP19A1. It results in disorders of sex development and skeletal malformations similar to Antley-Bixley syndrome. We report the case of a 9-year-old girl who was born with virilized genitalia (Prader stage V), absence of palpable gonads, 46,XX karyotype, and hypergonadotropic hypogonadism. During the first year of life, ovarian cyst, partial adrenal insufficiency, and osteoarticular changes, such as mild craniosynostosis, carpal and tarsal synostosis, and limited forearm pronosupination were observed. Her mother presented severe virilization during pregnancy. The molecular analysis of P450 oxidoreductase gene revealed compound heterozygosis for the nonsense p.Arg223*, and the novel missense p.Met408Lys, inherited from the father and the mother, respectively. © ABEM todos os direitos reservados.568578585Miller, W.L., Minireview: Regulation of steroidogenesis by electron transfer (2005) Endocrinology, 146, pp. 2544-2550Scott, R.R., Gomes, L.G., Huang, N., Van Vliet, G., Miller, W.L., Apparent manifesting heterozygosity in P450 oxidoreductase deficiency and its effect on coexisting 21-hydroxylase deficiency (2007) J Clin Endocrinol Metab, 92, pp. 2318-2322Auchus, R.J., Lee, T.C., Miller, W.L., Cytochrome b5 augments the 17,20-lyase activity of human P450c17 without direct electron transfer (1998) J Biol Chem, 273, pp. 3158-3165Flück, C.E., Tajima, T., Pandey, A.V., Arlt, W., Okuhara, K., Verge, C.F., Mutant P450 oxidoreductase causes disordered steroidogenesis with and without Antley-Bixler syndrome (2004) Nat Genet, 36, pp. 228-230Arlt, W., Walker, E.A., Draper, N., Ivison, H.E., Ride, J.P., Hammer, F., Congenital adrenal hyperplasia caused by mutant P450 oxidoreductase and human androgen synthesis: Analytical study (2004) Lancet, 363, pp. 2128-2135Scott, R.R., Miller, W.L., Genetic and clinical features of P450 oxidoreductase deficiency (2008) Horm Res, 69, pp. 266-275Polusani, S.R., Kar, R., Riquelme, M.A., Masters, B.S., Panda, S.P., Regulation of gap junction function and connexin 43 expression by cytochrome P450 oxidoreductase (CYPOR) (2011) Biochem Biophys Res Commun, 411, pp. 490-495Tomalik-Scharte, D., Maiter, D., Kirchheiner, J., Ivison, H.E., Fuhr, U., Arlt, W., Impaired hepatic drug and steroid metabolism in congenital adrenal hyperplasia due to P450 oxidoreductase deficiency (2010) Eur J Endocrinol, 163, pp. 919-924(2012) Human Gene Mutation Database, , http://www.hgmd.cf.ac.uk/ac/index.php, Available at, Accessed on: Nov 17Flück, C.E., Pandey, A.V., Clinical and biochemical consequences of P450 oxidoreductase deficiency (2011) Endocr Dev, 20, pp. 63-79Krone, N., Reisch, N., Idkowiak, J., Dhir, V., Ivison, H.E., Hughes, B.A., Genotype-phenotype analysis in congenital adrenal hyperplasia due to P450 oxidoreductase deficiency (2012) J Clin Endocrinol Metab, 97, pp. E257-E267Sambrook, J., Fristsch, E.F., Maniatis, T.E., (1989) Molecular Cloning: A laboratory manual, , Cold Spring Harbor, NY: Cold Spring Harbor Laboratory PressXia, C., Panda, S.P., Marohnic, C.C., Martásek, P., Masters, B.S., Kim, J.J., Structural basis for human NADPH-cytochrome P450 oxidoreductase deficiency (2011) Proc Natl Acad Sci U S A, 108, pp. 13486-13491Idkowiak, J., O'Riordan, S., Reisch, N., Malunowicz, E.M., Collins, F., Kerstens, M.N., Pubertal presentation in seven patients with congenital adrenal hyperplasia due to P450 oxidoreductase deficiency (2011) J Clin Endocrinol Metab, 96 (3), pp. E453-E462Hamdane, D., Xia, C., Im, S.C., Zhang, H., Kim, J.J., Waskell, L., Structure and function of an NADPH-cytochrome P450 oxidoreductase in an open conformation capable of reducing cytochrome P450 (2009) J Biol Chem, 284 (17), pp. 11374-11384Xia, C., Hamdane, D., Shen, A.L., Choi, V., Kasper, C.B., Pearl, N.M., Conformational changes of NADPH-cytochrome P450 oxidoreductase are essential for catalysis and cofactor binding (2011) J Biol Chem, 286, pp. 16246-16260Herkert, J.C., Blaauwwiekel, E.E., Hoek, A., Veenstra-Knol, H.E., Kema, I.P., Arlt, W., A rare cause of congenital adrenal hyperplasia: Antley-Bixler syndrome due to POR deficiency (2011) Neth J Med, 69, pp. 281-283Fukami, M., Hasegawa, T., Horikawa, R., Ohashi, T., Nishimura, G., Homma, K., Cytochrome P450 oxidoreductase deficiency in three patients initially regarded as having 21-hydroxylase deficiency and/or aromatase deficiency: Diagnostic value of urine steroid hormone analysis (2006) Pediatr Res, 59, pp. 276-280Flück, C.E., Miller, W.L., P450 oxidoreductase deficiency: A new form of congenital adrenal hyperplasia (2006) Curr Opin Pediatr, 18, pp. 435-441Homma, K., Hasegawa, T., Nagai, T., Adachi, M., Horikawa, R., Fujiwara, I., Urine steroid hormone profile analysis in cytochrome P450 oxidoreductase deficiency: Implication for the backdoor pathway to dihydrotestosterone (2006) J Clin Endocrinol Metab, 91, pp. 2643-2649But, W.M., Lo, I.F., Shek, C.C., Tse, W.Y., Lam, S.T., Ambiguous genitalia, impaired steroidogenesis, and Antley-Bixler syndrome in a patient with P450 oxidoreductase deficiency (2010) Hong Kong Med J, 16, pp. 59-62Iijima, S., Ohishi, A., Ohzeki, T., Cytochrome P450 oxidoreductase deficiency with Antley-Bixler syndrome: Steroidogenic capacities (2009) J Pediatr Endocrinol Metab, 22, pp. 469-475Ko, J.M., Cheon, C.K., Kim, G.H., Yoo, H.W., A case of Antley-Bixler syndrome caused by compound heterozygous mutations of the cytochrome P450 oxidoreductase gene (2009) Eur J Pediatr, 168, pp. 877-880Sahakitrungruang, T., Huang, N., Tee, M.K., Agrawal, V., Russell, W.E., Crock, P., Clinical, genetic, and enzymatic characterization of P450 oxidoreductase deficiency in four patients (2009) J Clin Endocrinol Metab, 94, pp. 4992-5000Adachi, M., Tachibana, K., Asakura, Y., Yamamoto, T., Hanaki, K., Oka, A., Compound heterozygous mutations of cytochrome P450 oxidoreductase gene (POR) in two patients with Antley-Bixler syndrome (2004) Am J Med Genet A, 128, pp. 333-339New, M.I., Nonclassical congenital adrenal hyperplasia and the polycystic ovarian syndrome (1993) Ann NY Acad Sci, 687, pp. 193-205Rosa, S., Duff, C., Meyer, M., Lang-Muritano, M., Balercia, G., Boscaro, M., P450c17 deficiency: Clinical and molecular characterization of six patients (2007) J Clin Endocrinol Metab, 92, pp. 1000-1007Shima, M., Tanae, A., Miki, K., Katsumata, N., Matsumoto, S., Nakajima, S., Mechanism for the development of ovarian cysts in patients with congenital lipoid adrenal hyperplasia (2000) Eur J Endocrinol, 142, pp. 274-279Belgorosky, A., Pepe, C., Marino, R., Guercio, G., Saraco, N., Vaiani, E., 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Supplementary Material for: Functional Impact of Novel Androgen Receptor Mutations on the Clinical Manifestation of Androgen Insensitivity Syndrome

<p>Androgens are responsible for the development and maintenance of male sex characteristics. Dysfunctions in androgen action due to mutations in the androgen receptor gene (<i>AR</i>) can lead to androgen insensitivity syndrome (AIS) that can be classified as mild (MAIS), partial (PAIS), or complete (CAIS). We have analyzed functional effects of p.Ser760Thr, p.Leu831Phe, p.Ile899Phe, p.Leu769Val, and p.Pro905Arg mutations and the combination p.Gln799Glu + p.Cys807Phe that were identified in patients with PAIS or CAIS. The p.Leu769Val and p.Pro905Arg mutations showed complete disruption of AR action under physiological hormone concentrations; however, they differed in high DHT concentrations especially in the N/C terminal interaction assay. Mutations p.Ser760Thr, p.Leu831Phe, p.Ile899Phe presented transactivation activities higher than 20% of the wild type in physiological hormone concentrations and increased with higher DHT concentrations. However, each one showed a different profile in the N/C interaction assay. When p.Gln799Glu and p.Cys807Phe were analyzed in combination, transactivation activities <10% in physiologic hormone conditions indicated an association with a CAIS phenotype. We conclude that the functional analysis elucidated the role of mutant ARs, giving clues for the molecular mechanisms associated with different clinical AIS manifestations. Differences in hormone-dependent profiles may provide a basis for the response to treatment in each particular case.</p

Níveis séricos de hemoglobina em adolescentes segundo estágio de maturação sexual Hemoglobin serum levels in adolescents according to sexual maturation stage

A adolescência constitui etapa de risco para o desenvolvimento da anemia ferropriva, uma vez que ocorre aumento da necessidade de ferro decorrente do crescimento estatural e da maturação biológica. Estudaram-se 130 adolescentes, de ambos os sexos, para verificar os valores de hemoglobina sérica em diferentes fases de maturação sexual. Utilizou-se o método de fotometria para dosar a hemoglobina sérica e realizou-se auto-avaliação do estágio de maturação sexual com base nos critérios de Tanner. Os níveis médios de hemoglobina foram semelhantes entre sexos, bem como entre meninas que menstruavam ou não. O nível médio de hemoglobina foi de 13,3g/dL tanto para os meninos como para as meninas (p=0,64), com desvios-padrão de 1,12 e de 0,83, respectivamente. Entre os adolescentes estudados, 7,7% tinham anemia ferropriva. Recomenda-se atenção ao grupo de adolescentes, devido ao aumento da necessidade de ferro durante o estirão de crescimento, principalmente entre as meninas, aumentando a suscetibilidade à anemia.<br>During adolescence, the risk of development of iron-deficiency anemia is higher because of the growth spurt and the sexual maturation which increase the iron requirement. One hundred and thirty adolescents (males and females) were studied, in order to assess the serum hemoglobin values in different sexual maturation stages. The photometric method was used and a self-evaluation of the sexual maturation stage based on Tanner's criteria was applied. The average hemoglobin values were similar for both sexes, as well for girls who had menstruated or not. The average hemoglobin values were 13.3 (s.d. 1.12)g/dL for males and 13.3 (s.d. 0.83)g/dL for females (p=0.64). Among the studied adolescents, 7.7% had iron-deficiency anemia. Due to the iron requirement increase during the growth spurt, mainly in females, and the higher susceptibility to iron deficiency anemia, special attention to the adolescents is recommended