RESUMO
BACKGROUND: Overexpression of the oncogene yes-associated-protein-1 (YAP1) is associated with increased cell proliferation in human cancers. YAP1 is a potential target of the Wnt/beta-catenin pathway, which plays an important role in adrenocortical tumors (ACT). The role of YAP1 in adrenocortical tumorigenesis has not been assessed. AIMS: To evaluate YAP1 expression in normal adrenals and pediatric ACT and its association with disease outcome. To investigate the interaction between YAP1 and the Wnt/beta-catenin pathway in adrenocortical cells. RESULTS: Strong YAP1 staining was present in fetal adrenals and pediatric ACT but weak in postnatal adrenals. In pediatric ACT, YAP1 mRNA overexpression was associated with death, recurrent/metastatic disease and lower overall survival. The inhibition of the Wnt/beta-catenin pathway increased YAP1 mRNA expression. siYAP1 increased CTNNB1/beta-catenin expression and nuclear staining regardless of DLV2, moreover, it decreased cell growth and impaired cell migration. MATERIALS AND METHODS: We assessed in 42 pediatric ACT samples the YAP1 protein expression by immunohistochemistry and mRNA expression by RT-qPCR and analyzed their association with outcome. As controls, we resort 32 fetal and postnatal normal adrenals for IHC and 10 normal adrenal cortices for RT-qPCR. The interaction between YAP1 and the Wnt/beta-catenin pathway was assessed in NCI-H295 adrenocortical cells by inhibiting the TCF/beta-catenin complex and by knocking down YAP1. CONCLUSION: YAP1 overexpression is a marker of poor prognosis for pediatric patients with ACT. In adrenocortical cells, there is a close crosstalk between YAP1 and Wnt/beta-catenin. These data open the possibility of future molecular therapies targeting Hippo/YAP1 signaling to treat advanced ACT.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Neoplasias do Córtex Suprarrenal/metabolismo , Córtex Suprarrenal/metabolismo , Fosfoproteínas/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Adolescente , Neoplasias do Córtex Suprarrenal/genética , Neoplasias do Córtex Suprarrenal/mortalidade , Carcinogênese , Linhagem Celular Tumoral , Proliferação de Células , Criança , Pré-Escolar , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Lactente , Masculino , Metástase Neoplásica , Recidiva Local de Neoplasia , Fosfoproteínas/genética , Transdução de Sinais , Análise de Sobrevida , Fatores de Transcrição , Proteínas Wnt/metabolismo , Proteínas de Sinalização YAP , beta Catenina/metabolismoRESUMO
STUDY OBJECTIVES: To evaluate bone quantity and quality in postmenarchal adolescents treated for idiopathic central precocious puberty (CPP) in childhood with a gonadotropin-releasing hormone analog (GnRHa) and to determine the serum concentrations of bone remodeling markers. DESIGN AND PARTICIPANTS: This cross-sectional study included 53 postmenarchal adolescent girls who were divided into 2 groups: 27 adolescents who were treated with GnRHa in childhood for idiopathic CPP (the CPP group) and 26 women who presented with physiological development of secondary sex traits (the control group). INTERVENTIONS: None. MAIN OUTCOME MEASURES: Weight, height, body mass index, age at menarche, time since menarche, body composition, bone mineral density (BMD), bone quality, and serum insulin, glucose, osteocalcin, and carboxyl-terminal telopeptide of type I collagen concentrations were compared in the 2 groups. BMD data were analyzed by using both dual-energy x-ray absorptiometry (DXA) and osteosonography, and body composition was measure with the use of DXA and electrical bioimpedance. RESULTS: BMD and bone quality did not differ significantly between the CPP and control groups when analyzed by using DXA or osteosonography. Serum osteocalcin concentration was significantly lower (P = .02) in the CPP than in the control group. Insulin was higher in the CPP group, and hyperinsulinemia was an independent predictor of bone quantity and quality assessed by using osteosonography. Body mass index and percent fat were determined by using DXA, and the duration of use of GnRHa treatment and the time since GnRHa discontinuation were not independent predictors of bone quantity and quality. CONCLUSION: Postmenarchal adolescents treated with GnRHa for CPP in childhood did not show a reduction in bone quantity or quality.
Assuntos
Densidade Óssea/efeitos dos fármacos , Remodelação Óssea/efeitos dos fármacos , Hormônio Liberador de Gonadotropina/análogos & derivados , Puberdade Precoce/tratamento farmacológico , Absorciometria de Fóton , Adolescente , Biomarcadores/sangue , Composição Corporal , Estatura , Índice de Massa Corporal , Peso Corporal , Criança , Estudos Transversais , Feminino , Hormônio Liberador de Gonadotropina/efeitos adversos , Hormônio Liberador de Gonadotropina/uso terapêutico , Humanos , MenarcaRESUMO
CONTEXT AND OBJECTIVE: Sonic Hedgehog (SHH) and GLI2, an obligatory mediator of SHH signal transduction, are holoprosencephaly (HPE)-associated genes essential in pituitary formation. GLI2 variants have been found in patients with congenital hypopituitarism without complex midline cerebral defects (MCD). However, data on the occurrence of SHH mutations in these patients are limited. We screened for SHH and GLI2 mutations or copy number variations (CNV) in patients with congenital hypopituitarism without MCD or with variable degrees of MCD. PATIENTS AND METHODS: Detailed data on clinical, laboratory and neuroimaging findings of 115 patients presenting with congenital hypopituitarism without MCD, septo-optic dysplasia or HPE were analysed. The SHH and GLI2 genes were directly sequenced, and the presence of gene CNV was analysed by multiplex ligation-dependent probe amplification (MLPA). RESULTS: Anterior pituitary deficiency was found in 74% and 53% of patients with SOD or HPE, respectively. Diabetes insipidus was common in patients with HPE (47%) but infrequent in patients with congenital hypopituitarism or SOD (7% and 8%, respectively). A single heterozygous nonsense SHH mutation (p.Tyr175Ter) was found in a patient presenting with hypopituitarism and alobar HPE. No other SHH mutations or CNV were found. Nine GLI2 variations (8 missense and 1 frameshift) including a homozygous and a compound heterozygous variation were found in patients with congenital hypopituitarism or SOD, but not in HPE patients. No GLI2 CNV were found. CONCLUSION: SHH mutations or copy number variations are not a common cause of congenital hypopituitarism in patients without complex midline cerebral defects. GLI2 variants are found in some patients with congenital hypopituitarism without complex midline cerebral defects or septo-optic dysplasia. However, functional analyses of these variants are needed to strengthen genotype-phenotype relationship.
Assuntos
Proteínas Hedgehog/genética , Hipopituitarismo/congênito , Hipopituitarismo/genética , Mutação , Adolescente , Adulto , Encéfalo/fisiopatologia , Criança , Pré-Escolar , Feminino , Dosagem de Genes , Estudos de Associação Genética , Variação Genética , Heterozigoto , Holoprosencefalia/genética , Humanos , Lactente , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Imageamento por Ressonância Magnética , Masculino , Mutação de Sentido Incorreto , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fenótipo , Hipófise/metabolismo , Transdução de Sinais , Adulto Jovem , Proteína Gli2 com Dedos de ZincoRESUMO
BACKGROUND: Low expression of HLA class II antigens has been associated with more aggressive disease in several human malignancies including adult adrenocortical tumors (ACT), but their clinical relevance in pediatric ACT needs to be investigated. PROCEDURE: This study analyzed the expression profile of three class II histocompatibility genes (HLA-DRA, HLA-DPA1, and HLA-DPB1) in 58 consecutive pediatric ACT (13 adenomas and 45 carcinomas) by quantitative real time PCR and their association with clinical and biological features. HLA-DPA1 protein level was determined by immunohistochemistry. RESULTS: A significant association (P < 0.01) was observed between lower expression levels of the three genes analyzed and poor prognostic factors such as age ≥ 4 years, tumor size ≥ 200 cm(3), tumor weight ≥ 100 g, and metastatic disease; the presence of an unfavorable event and death. Underexpression of the HLA-DRA, HLA-DPA1, and HLA-DPB1 genes were associated with lower 5-year event-free survival (EFS) (P = 0.017, P < 0.001, and P = 0.017, respectively). Cox multivariate analysis showed that HLA-DPA1 was an independent prognostic factor (P = 0.029) when analyzed in association with stage IV, age and tumor size. Significantly lower EFS was also observed in patients with negative/weak immunostaining for HLA-DPA1 (P = 0.002). Similar results were observed when only patients classified as having carcinomas were analyzed. CONCLUSION: Our results suggest that lower expression of HLA-DRA, HLA-DPA1, and HLA-DPB1 genes may contribute to more aggressive disease in pediatric ACT. HLA-DPA1 immunostaining may represent potential aggressiveness marker in this tumor.
Assuntos
Neoplasias do Córtex Suprarrenal/imunologia , Cadeias alfa de HLA-DP/genética , Cadeias beta de HLA-DP/genética , Cadeias alfa de HLA-DR/genética , Adolescente , Neoplasias do Córtex Suprarrenal/mortalidade , Neoplasias do Córtex Suprarrenal/patologia , Criança , Pré-Escolar , Feminino , Cadeias alfa de HLA-DP/análise , Cadeias beta de HLA-DP/análise , Cadeias alfa de HLA-DR/análise , Humanos , Lactente , Masculino , PrognósticoRESUMO
BACKGROUND: The sonic hedgehog (SHH) pathway plays a key role in rodent adrenal cortex development and is involved in tumorigenesis in several human tissues, but data in human adrenal glands are limited. OBJECTIVES: The objectives of the study were to analyze the involvement of the SHH pathway in human adrenal development and tumorigenesis and the effects of SHH inhibition on an adrenocortical tumor (ACT) cell line. PATIENTS AND METHODS: Expression of SHH pathway components was evaluated by immunohistochemistry in 51 normal adrenals (33 fetal) and 34 ACTs (23 pediatric) and by quantitative PCR in 81 ACTs (61 pediatric) and 19 controls (10 pediatric). The effects of SHH pathway inhibition on gene expression and cell viability in the NCI-H295A adrenocortical tumor cell line after cyclopamine treatment were analyzed. RESULTS: SHH pathway proteins were present in fetal and postnatal normal adrenals and showed distinct patterns of spatiotemporal expression throughout development. Adult adrenocortical carcinomas presented with higher expression of PTCH1, SMO, GLI3, and SUFU compared with normal adult adrenal cortices. Conversely, pediatric ACTs showed lower mRNA expression of SHH, PTCH1, SMO, GLI1, and GLI3 compared with normal pediatric adrenal cortices. In vitro treatment with cyclopamine resulted in decreased GLI3, SFRP1, and CTNNB1 mRNA expression and ß-catenin staining as well as decreased cell viability. CONCLUSIONS: The SHH pathway is active in human fetal and postnatal adrenals, up-regulated in adult adrenocortical carcinomas, and down-regulated in pediatric ACTs. SHH pathway antagonism impaired cell viability. The SHH pathway is deregulated in ACTs and might provide a new target therapy to be explored.
Assuntos
Neoplasias do Córtex Suprarrenal/genética , Córtex Suprarrenal/embriologia , Córtex Suprarrenal/crescimento & desenvolvimento , Carcinoma Adrenocortical/genética , Proteínas Hedgehog/genética , Córtex Suprarrenal/metabolismo , Neoplasias do Córtex Suprarrenal/metabolismo , Carcinoma Adrenocortical/metabolismo , Adulto , Estudos de Casos e Controles , Células Cultivadas , Criança , Desenvolvimento Embrionário/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Proteínas Hedgehog/metabolismo , Humanos , Recém-Nascido , Gravidez , Estudos Retrospectivos , Transdução de Sinais/genéticaRESUMO
CONTEXT: The role of planar cell polarity (Wnt/PCP) and calcium-dependent (Wnt/Ca) noncanonical Wnt pathways in adrenocortical tumours (ACTs) is unknown. OBJECTIVES: To investigate the gene expression of Wnt/PCP and Wnt/Ca pathways and its association with TP53 p.R337H and CTNNB1 mutations in paediatric and adult ACTs and to correlate these findings with clinical outcome. PATIENTS: Expression of noncanonical Wnt-related genes was evaluated in 91 ACTs (66 children and 25 adults) by qPCR and the expression of beta-catenin, P53 and protein effectors of Wnt/Ca (NFAT) and Wnt/PCP (JNK) by immunohistochemistry. TP53 and CTNNB1 genes were sequenced. RESULTS: TP53 p.R337H mutation frequency was higher in children (86% vs 28%), while CTNNB1 mutation was higher in adults (32% vs 6%). Mortality was higher in adults harbouring TP53 p.R337H and in children with CTNNB1 mutations. Overexpression of WNT5A, Wnt/Ca ligand, was observed in children and adults. Overexpression of MAPK8 and underexpression of PRICKLE, Wnt/PCP mediators, were observed in paediatric but not in adult cases. Cytoplasmic/nuclear beta-catenin and P53 accumulation was observed in the majority of paediatric and adult ACTs as well as NFAT and JNK. Overexpression of MAPK8 and underexpression of PRICKLE were associated with mortality in children, while overexpression of WNT5A and underexpression of PRICKLE were associated with mortality in adults. CONCLUSIONS: In our study, TP53 p.R337H and CTNNB1 mutations correlated with poor prognosis in adults and children, respectively. We demonstrate, for the first time, the activation of Wnt/PCP and Wnt/Ca noncanonical pathway genes, and their association with poor outcome in children and adults, suggesting their putative involvement in ACTs aggressiveness.
Assuntos
Neoplasias do Córtex Suprarrenal/metabolismo , Via de Sinalização Wnt/fisiologia , Adolescente , Neoplasias do Córtex Suprarrenal/genética , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Imuno-Histoquímica , Lactente , Masculino , Pessoa de Meia-Idade , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Via de Sinalização Wnt/genética , Adulto Jovem , beta Catenina/genética , beta Catenina/metabolismoRESUMO
OBJECTIVES: Idiopathic central precocious puberty and its postponement with a (gonadotropin-releasing hormone) GnRH agonist are complex conditions, the final effects of which on bone mass are difficult to define. We evaluated bone mass, body composition, and bone remodeling in two groups of girls with idiopathic central precocious puberty, namely one group that was assessed at diagnosis and a second group that was assessed three years after GnRH agonist treatment. METHODS: The precocious puberty diagnosis and precocious puberty treatment groups consisted of 12 girls matched for age and weight to corresponding control groups of 12 (CD) and 14 (CT) girls, respectively. Bone mineral density and body composition were assessed by dual X-ray absorptiometry. Lumbar spine bone mineral density was estimated after correction for bone age and the mathematical calculation of volumetric bone mineral density. CONEP: CAAE-0311.0.004.000-06. RESULTS: Lumbar spine bone mineral density was slightly increased in individuals diagnosed with precocious puberty compared with controls; however, after correction for bone age, this tendency disappeared (CD = -0.74 + 0.9 vs. precocious puberty diagnosis = -1.73 + 1.2). The bone mineral density values of girls in the precocious puberty treatment group did not differ from those observed in the CT group. CONCLUSION: There is an increase in bone mineral density in girls diagnosed with idiopathic central precocious puberty. Our data indicate that the increase in bone mineral density in girls with idiopathic central precocious puberty is insufficient to compensate for the marked advancement in bone age observed at diagnosis. GnRH agonist treatment seems to have no detrimental effect on bone mineral density.
Assuntos
Composição Corporal/fisiologia , Densidade Óssea/fisiologia , Hormônio Liberador de Gonadotropina/agonistas , Puberdade Precoce/tratamento farmacológico , Puberdade Precoce/patologia , Absorciometria de Fóton , Adolescente , Fatores Etários , Composição Corporal/efeitos dos fármacos , Índice de Massa Corporal , Densidade Óssea/efeitos dos fármacos , Estudos de Casos e Controles , Criança , Feminino , Humanos , Valores de Referência , Estatísticas não Paramétricas , Resultado do Tratamento , Adulto JovemRESUMO
CONTEXT: CTNNB1/ß-catenin mutations and activation of Wnt/ß-catenin pathway are frequent in adult adrenocortical tumors (ACT), but data on childhood ACT are lacking. OBJECTIVE: The aim of the study was to investigate the presence of Wnt/ß-catenin pathway abnormalities in childhood ACT. PATIENTS AND METHODS: Clinicopathological findings and outcome of 62 childhood ACT patients were analyzed regarding CTNNB1 mutations and the expression of Wnt-related genes (CTNNB1; WNT4, a Wnt ligand; SFRP1, DKK3, and AXIN1, Wnt inhibitors; TCF7, a transcription factor; and MYC and WISP2, target genes) by quantitative PCR and immunohistochemistry. RESULTS: CTNNB1-activating mutations were found in only four of 62 ACT (6%), all of them harboring TP53 mutation. There was association between the presence of CTNNB1 mutations and death (P = 0.02). Diffuse ß-catenin accumulation was found in 71% of ACT, even in ACT without CTNNB1 mutations. Compared to normal adrenals, ACT presented increased expression of CTNNB1 (P = 0.008) and underexpression of Wnt inhibitor genes: DKK3 (P < 0.0001), SFRP1 (P = 0.05), and AXIN1 (P = 0.04). With regard to Wnt/ß-catenin target genes, ACT presented increased expression of WISP2 but lower expression of MYC. Higher overall survival was associated with underexpression of SFRP1 (P = 0.01), WNT4 (P = 0.004), and TCF7 (P < 0.01). CONCLUSIONS: CTNNB1 mutations are not common in childhood ACT but appear to associate with poor prognosis. Nevertheless, most ACT exhibit increased expression of ß-catenin and WISP2 and reduced expression of Wnt inhibitor genes (DKK3, SFRP1, and AXIN1). Thus, in addition to CTNNB1 mutations, other genetic events affecting the Wnt/ß-catenin pathway may be involved in childhood adrenocortical tumorigenesis.
Assuntos
Neoplasias do Córtex Suprarrenal/metabolismo , Transdução de Sinais/fisiologia , Proteínas Wnt/fisiologia , beta Catenina/fisiologia , Adolescente , Neoplasias do Córtex Suprarrenal/genética , Proteína Axina/fisiologia , Proteínas de Sinalização Intercelular CCN , Criança , Pré-Escolar , Estudos de Coortes , DNA/genética , DNA/isolamento & purificação , Feminino , Humanos , Imuno-Histoquímica , Lactente , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Masculino , Mutação/fisiologia , Proteínas Proto-Oncogênicas c-myc/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/isolamento & purificação , Proteínas Repressoras , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/genética , Análise de Sobrevida , Fator 1 de Transcrição de Linfócitos T/fisiologia , Fatores de Transcrição/fisiologia , Resultado do Tratamento , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteínas Wnt/antagonistas & inibidores , Proteínas Wnt/genética , Proteína Wnt4/fisiologia , beta Catenina/genéticaRESUMO
Gómez-López-Hernández (GLH) syndrome or cerebello-trigeminal dysplasia is a neurocutaneous syndrome whose etiology is unknown at the present time. We report two additional Brazilian patients, including the oldest one known to date (age 29). Here, we review the expanded phenotype in four patients with new clinical, psychiatric, radiological, and molecular investigations. One patient may have hypomania within the bipolar spectrum disorder with onset in childhood and adolescence. Primary growth hormone (GH) deficiency was ruled out in all patients, although one of them might have developed secondary GH deficiency due to partial hypopituitarism following severe hydrocephalus. Brain magnetic resonance angiography disclosed no azygous anterior cerebral artery (ACA) but only normal variants. Molecular analysis of the lysosomal acid phosphatase gene (ACP2) was performed, but no pathogenic mutations were identified. We present an overview of the phenotypic features of all patients described to date. There are currently 12 unrelated patients reported in the literature, 5 of whom are Brazilian. We discuss new molecular insights and speculate about the pathogenesis of GLH syndrome.
Assuntos
Anormalidades Múltiplas/diagnóstico , Anormalidades Múltiplas/genética , Síndromes Neurocutâneas/diagnóstico , Síndromes Neurocutâneas/genética , Anormalidades Múltiplas/patologia , Adulto , Alopecia/patologia , Brasil , Cerebelo/anormalidades , Criança , Transtornos do Crescimento/patologia , Humanos , Masculino , Síndromes Neurocutâneas/patologia , Fenótipo , SíndromeRESUMO
OBJECTIVES: To identify the prevalence of iron deficiency in the population studied, as well as verifying if such deprivation is associated with vitamin A deficiency. METHODS: One hundred seventy-nine children, > or = 24 months and < 72 months of age, with no diarrhea and/or fever at collection were studied. Vitamin A deficiency identification was carried out through serum 30-day dose-response test. Samples of peripheral blood from fasting children was obtained for hemoglobin counts, serum iron, and unsaturated iron binding capacity assays. Information about the presence of diarrhea and/or fever during the 15 days preceding the study was also obtained. RESULTS: 35.8% (64/179) of the children presented iron deficiency and 75.4% (135/179), vitamin A deficiency. 29.1% (52/179) of the children presented both iron and vitamin A deficiencies. Iron deficiency was not associated with vitamin A deficiency. A separate analysis for each hematimetric index also demonstrated no significant difference between children with or without vitamin A deficiency. Children aged 24 to 36 months presented significantly higher prevalence rates of iron deficiency (p = 0.0005) as did children with diarrhea and/or fever during the 15 days preceding the study (p = 0.003). CONCLUSIONS: Although iron deficiency was not associated with vitamin A deficiency, high rates of both deficiencies were exhibited in a "healthy" population with low malnutrition indices. Such situations are known as "hidden hunger". Younger children presented a higher risk of iron deficiency as did children with diarrhea and/or fever during the 15 days preceding the study.
Assuntos
Anemia Ferropriva/epidemiologia , Deficiência de Vitamina A/epidemiologia , Anemia Ferropriva/complicações , Brasil/epidemiologia , Criança , Pré-Escolar , Diarreia/epidemiologia , Feminino , Febre/epidemiologia , Humanos , Masculino , Prevalência , Deficiência de Vitamina A/complicaçõesRESUMO
Premature thelarche (PT) is characterised by precocious breast development without any other sign of puberty, normal height velocity (HV) and normal bone maturation, while girls with central precocious puberty (CPP) show increased HV, bone maturation and increased serum IGF-I and IGFBP-3 levels. This prompted us to study serum IGF-I and IGFBP-3 concentrations in girls with PT. Thirty-nine girls with premature breast development were studied and classified as PT or CPP according to clinical and laboratory evaluation. Normal prepubertal and pubertal girls were studied as controls. Serum IGF-I and IGFBP-3 were determined in all girls by IRMA. IGF-I levels in PT (155 +/- 61 microg/l) were lower than in CPP (337 +/- 149 microg/l) or late-pubertal controls (355 +/- 84 microg/l) and similar to those found in prepubertal (113 +/- 72 microg/l) and early-pubertal (222 +/- 81 microg/l) girls. Considering the SDS of IGF-I for chronological age (CA), the values observed in PT were in an intermediate position between CPP and prepubertal controls and statistically similar to those observed in CPP and prepubertal girls. IGFBP-3 levels in PT (2.1 +/- 0.5 mg/l) were similar to those found in CPP (2.5 +/- 0.8 mg/l), but only the latter were higher than in prepubertal girls (1.9 +/- 0.9 mg/l). IGF-I/IGFBP-3 molar ratios in PT were in an intermediate position between CPP and prepubertal controls. In conclusion, IGF-I and IGF-I/IGFBP-3 values in PT are intermediate between those observed in prepubertal children and in CPP, suggesting that PT could be a very early stage of puberty with slight but real changes in the GH-IGF axis.