Abstract Background: Air pollution in industrial and urban centers is related to negative effects... more Abstract Background: Air pollution in industrial and urban centers is related to negative effects in living organisms, including alterations in male fertility. The fine inhalable particulate matter is the most deleterious fraction of air pollution and consists of a complex mixture of particles sized 2.5 µm or less of aerodynamic diameter (PM2.5). Experimental studies had already showed a deleterious effect of air pollution of Sao Paulo city on fertility in male mice. The hypothesis of its toxicity is based on activation of oxidative stress and inflammatory via as well as DNA adducts, leading to epigenetic alterations. Aim: To evaluate, under different conditions, the effects of the exposure to air pollution of city of Sao Paulo on mice testis. Methods: A group of mice was exposed during gestational period and after birth, from the weaning day until adulthood (prenatal and post-weaning group - PNPWG); another group of mice was exposed to PM2.5 during gestational period only (prenatal group - PNG); a third group of mice was exposed to PM2.5 after birth, from the weaning day until adulthood only (post-weaning group - PWG); and finally, a fourth group of mice was exposed to filtered air during gestational period and from post-weaning day until adulthood (control group - CG). Analyzes were performed on testis from adult animals. Stereological techniques were used to analyze structures of testis and Johnsen's score to evaluate spermatogenesis in a qualitative way. DNA microarray were used to evaluate gene expression. Results: Testis of all the exposed groups showed tubular degeneration characteristics. In comparison with CG, the testis of PNPWG showed higher weight and volume, surface area of the seminiferous tubules, volume of the seminiferous epithelium and epididymis weight. Quality of spermatogenesis in PNPWG was also significantly worse in comparison with CG. PWG had similar results when compared with CG, however the only parameters with significantly difference were the volume of the seminiferous epithelium and the quality of spermatogenesis. Mean volume of the seminiferous epithelium from PNG was significantly lower when compared to CG. Gene expression analysis exhibited differential expression of genes mostly related to mitochondrial tRNA in group-to-group comparison. The mt-Ty (mitochondrially encoded tRNA tyrosine) gene was hiperexpressed in PNPWG in comparison with other groups. Conclusion: We demonstrated for the first time that the exposure to PM2.5 of the city of Sao Paulo causes tubular degeneration with decreased or absent spermatogenesis associated with differential expression of genes related to mitochondrial function in mice testis. In addition, we demonstrated that post-natal exposure seems to be more deleterious for the testicular function.
A sindrome de Kallmann (SK) e uma afeccao relativamente rara, caracterizada pela associacao de hi... more A sindrome de Kallmann (SK) e uma afeccao relativamente rara, caracterizada pela associacao de hipogonadismo hipogonadotrofico (HH) e anosmia/hiposmia. Embora a maioria dos casos de SK seja esporadica, diferentes padroes de heranca, recessivo ligado ao X, autossomico dominante e autossomico recessivo tem sido descritos, sendo que a forma ligada ao X da sindrome (SKX) encontra-se genotipica e fenotipicamente melhor definida. O gene KAL-1 localizado em Xp22.3 escapa a inativacao e codifica uma proteina, a anosmina, cujo papel tem sido relacionado ao processo de migracao de neuronios produtores de GnRH que ocorre durante a embriogenese humana. No entanto, muitos casos de hipogonadismo hipogonadotrofico nao podem ser relacionados a qualquer alteracao olfativa, sendo estes classificados como hipogonadismo hipogonadotrofico normosmicos (HHn). Mutacoes no gene do receptor do hormonio liberador de gonadotrofinas (GnRH-R) tem sido descritas numa parcela desses casos. Alem disso, os genes NELF (fator nasal embrionario do hormonio liberador de hormonio luteinizante) e EBF2 (?early B-cell factor?) tem sido relacionados ao processo de migracao de neuronios produtores de GnRH, sendo considerados possivelmente envolvidos na genese do HH. O presente trabalho foi elaborado com o objetivo de se investigar a base genetica das diferentes formas do HH. Para tal, foram avaliados 31 individuos com hipogonadismo hipogonadotrofico (pertencentes a 26 familias), 22 com anosmia e nove com olfacao normal, mediante a analise do cariotipo pela tecnica de bandamento GTG seguida da analise molecular dos genes KAL-1 e GnRH-R, por PCR e sequenciamento. Todos os pacientes estudados apresentaram cariotipo normal, a excecao de um caso esporadico de SK com microdelecao do locus KAL-1 detectada somente por FISH em estudo anteriormente realizado. Entre os pacientes com SK, foram detectadas mutacoes na regiao codificante do gene KAL-1 nas tres familias com heranca recessiva ligada ao X e em um caso esporadico. Duas familias apresentaram duas delecoes intragenicas similares envolvendo os exons 5-10; na terceira familia detectou-se uma mutacao 721C>T no exon 5 levando a um codon de parada prematuro. Esta mutacao ainda nao se encontra descrita na literatura. No caso esporadico identificou-se uma mutacao ?frameshift? 1956delC no exon 12. Em relacao aos pacientes com HHn, nao foram identificadas mutacoes nos genes GnRH-R e KAL-1. Nesses casos, e nos pacientes com SK cuja etiologia nao pode ser esclarecida, foram ainda analisados os genes NELF e EBF2. Contudo, nenhuma alteracao nas sequencias desses genes foi identificada entre os pacientes da nossa casuistica. Em conclusao, nao foram encontradas evidencias cromossomicas da localizacao de outros genes candidatos responsaveis pelo surgimento do HH. Duas mutacoes novas e duas delecoes intragenicas semelhantes foram identificadas no gene KAL-1 entre os casos esporadicos e familiais da SK, observando-se que essas mutacoes ocorreram numa frequencia relativamente elevada em nossa casuistica. Os pacientes com HHn nao apresentaram mutacoes nos genes KAL-1 e GnRH-R. Nao foram observadas anomalias nas sequencias codificantes dos genes NELF e EBF2 em nenhuma das formas de HH Abstr
PurposeCushing’s disease (CD) is a severe illness generally caused by microcorticotropinomas (MIC... more PurposeCushing’s disease (CD) is a severe illness generally caused by microcorticotropinomas (MICs) and in approximately 7–20% of patients by macrocorticotropinomas (MACs). USP8-mutations have been identified as a major genetic cause of CD (~ 50%). Few studies have reported the distribution between MICs–MACs related to USP8-mutations and their genotype–phenotype correlations. Therefore, we aimed to evaluate USP8-mutations in a cohort of MICs–MACs from a unique center and to perform a systematic review and meta-analysis.MethodsDNA-tumor-tissues from 47 corticotropinomas (16 MICs and 31 MACs) were sequenced. Clinical-biochemical data, radiological imaging data and remission/recurrence rates were evaluated. In addition, we performed a meta-analysis of nine published series (n = 630).ResultsWe identified four different USP8-mutations previously described, in 11 out of 47 (23.4%) corticotropinomas; 8 out of 11 were MACs. The urinary cortisol levels of our patients with corticotrophin USP8-mutated-alleles were lower than those of patients with wild-type (WT) alleles (p ≤ 0.017). The frequency of USP8-mutated-alleles among the series was approximately 30% with a higher prevalence in female-patients (p < 0.1 × 10−4). Among the 5 series, the remission rates were higher in patients with USP8-mutated-alleles than in those with the USP8-WT-alleles (p < 0.1 × 10−4).ConclusionOur data, as well as the retrospective review of CD series associated with USP8-mutated alleles, show heterogeneous findings among the series. Several drawbacks included the lack of a systematic protocol to evaluate these patients before surgery and follow-up. Further prospective studies using a systematic protocol will provide more consistent information about the influence of the corticotropinomas with USP8-mutated alleles on the phenotype, responses to treatment and outcome of patients with CD.
Context Congenital hypogonadotropic hypogonadism (CHH) is a rare condition caused by GnRH deficie... more Context Congenital hypogonadotropic hypogonadism (CHH) is a rare condition caused by GnRH deficiency. Several genes have been associated with the pathogenesis of CHH, but most cases still remain without a molecular diagnosis. The advent of next-generation sequencing (NGS) has allowed the simultaneous genotyping of several regions, faster, making possible the extension of the genetic knowledge of CHH. Objective Genetic characterization of a large cohort of Brazilian CHH patients. Design and patients A cohort of 130 unrelated patients (91 males, 39 females) with CHH (75 normosmic CHH, 55 Kallmann syndrome) was studied using a panel containing 36 CHH-associated genes. Results Potential pathogenic or probably pathogenic variants were identified in 43 (33%) CHH patients. The genes ANOS1, FGFR1 and GNRHR were the most frequently affected. A novel homozygous splice site mutation was identified in the GNRH1 gene and a deletion of the entire coding sequence was identified in SOX10. Deleterio...
PurposeFilamin A (FLNA) expression is related to dopamine receptor type 2 (DRD2) expression in pr... more PurposeFilamin A (FLNA) expression is related to dopamine receptor type 2 (DRD2) expression in prolactinomas. Nevertheless, in corticotrophinomas, there are few studies about DRD2 expression and no data on FLNA. Therefore, we evaluated FLNA and DRD2 expression in corticotrophinomas and their association with tumor characteristics.MethodsDRD2 and FLNA expression by immunohistochemistry, using H-score, based on the percentage of positive cells in a continuous scale of 0–300, were evaluated in 23 corticotrophinomas samples from patients submitted to neurosurgery. In six patients, treatment with cabergoline was indicated after non curative surgery.ResultsTwenty-two patients were female and one male. Regarding tumor size, 10 were micro and 12 were macroadenomas. DRD2 expression was found in 89% of cases and did not correlate with FLNA expression. Moreover, the response to cabergoline, observed in 33% of the cases, did not correlate with DRD2 nor FLNA expression. FLNA expression was not associated with clinical and tumor characteristics, except for sphenoid sinus invasion.ConclusionsIn our cohort of corticotrophinomas, DRD2 expression was not associated with FLNA expression nor to the response to CAB. Nonetheless, FLNA expression could be related to tumor invasiveness.
Complete tumor removal by transsphenoidal surgery is usually difficult for large nonfunctioning p... more Complete tumor removal by transsphenoidal surgery is usually difficult for large nonfunctioning pituitary adenomas (NFPAs). A validated medical treatment may be useful for their management. This study evaluates the clinical efficacy of the dopaminergic agonist cabergoline for residual NFPA. We conducted a randomized, parallel, open-label clinical trial that compared cabergoline with nonintervention in patients with residual NFPA after transsphenoidal surgery over 2 years. The primary outcome was clinical efficacy (tumor reduction). The secondary outcome was the relationship between tumor dopamine D2 receptor (D2R) expression and clinical responsiveness. Tumor measurements and clinical evaluations were performed every 6 months. In total, 59 and 57 individuals were randomly assigned to the study and control groups, respectively. At the end of the study, residual tumor shrinkage, stabilization, and enlargement were observed in 28.8%, 66.1%, and 5.1% of patients, respectively, in the medical-therapy group and in 10.5%, 73.7%, and 15.8% of patients, respectively, in the control group (P=0.01). The progression-free survival rate was 23.2 and 20.8 months for the study and control groups, respectively (P=0.01). D2R was not associated with cabergoline responsiveness. No major side effects were related to cabergoline use. Cabergoline was an effective drug for treating residual NFPA, and its use was associated with a high rate of tumor shrinkage (ClinicalTrials.gov NCT03271918).
Tumor recurrence or incomplete resection in nonfunctioning pituitary adenomas (NFPAs) is relative... more Tumor recurrence or incomplete resection in nonfunctioning pituitary adenomas (NFPAs) is relatively common. However, predictive factors of tumor recurrence in NFPAs are not well established. We evaluated possible factors related to tumor recurrence in a large cohort of NFPAs at a single pituitary neurosurgery center. A retrospective analysis was conducted of 410 medical records of patients with NFPAs treated by transsphenoidal surgery between 2000 and 2014. Among the participants, 210 were female (51.0%). A total of 14.1% had giant adenomas. Null-cell pituitary adenomas (n = 239; 58.9%) were the most frequent, followed by silent gonadotroph adenomas (n = 112; 27.3%). Null-cell adenomas were more frequent in women (P = 0.008) and silent gonadotroph adenomas were more frequent in men (P = 0.004). Recurrence was not related to sex or age. Tumor recurrence occurred more often among silent corticotropic adenomas and giant adenomas (hazard ratio 2.45; P < 0.0001 and hazard ratio 2.35; ...
Sirtuins 1-7 (SIRT) are a highly conserved family of histone deacetylases involved in the regulat... more Sirtuins 1-7 (SIRT) are a highly conserved family of histone deacetylases involved in the regulation of longevity that have a considerable impact in transcription, DNA repair regulation, telomeric stability, cell senescence and apoptosis. In the present study, SIRT1-7 mRNA levels were evaluated in 37 somatotropinomas and 31 nonfunctioning pituitary adenomas (NFPAs) using qPCR and relation to tumor size, invasiveness and Ki-67 proliferative index was made. Overexpression of SIRT1 was observed in 86.5% of somatotropinomas versus 41.9% of NFPAs (P < 0.01). SIRT3 was more underexpressed in NFPAs than somatotropinomas (77.4 and 40.5%, respectively, P < 0.01) as well as SIRT4 and SIRT7. Despite the lack of association between sirtuins and invasiveness or Ki-67 index, SIRT1 and SIRT3 expressions were related to tumor size. Mean of the largest diameter was smaller in adenomas with SIRT1 overexpression than with normal expression (P < 0.01) and SIRT3 underexpression was associated w...
ACTH-dependent hypercortisolism caused by a pituitary adenoma [Cushing's disease (CD)] is the... more ACTH-dependent hypercortisolism caused by a pituitary adenoma [Cushing's disease (CD)] is the most common cause of endogenous Cushing's syndrome. CD is often associated with several morbidities, including hypertension, diabetes, osteoporosis/bone fractures, secondary infections, and increased cardiovascular mortality. While the majority (≈80%) of the corticotrophinomas visible on pituitary magnetic resonance imaging are microadenomas (MICs, <10 mm of diameter), some tumors are macroadenomas (MACs, ≥10 mm) with increased growth potential and invasiveness, exceptionally exhibiting malignant demeanor. In addition, larger and invasive MACs are associated with a significant increased risk of local complications, such as hypopituitarism and visual defects. Given the clinical and molecular heterogeneity of corticotrophinomas, the aim of this study was to investigate the pattern of genetic differential expression between MIC and MAC, including the invasiveness grade as a criterio...
Suppressor of cytokine signaling 2 (SOCS2) is a STAT5b-regulated gene and one of its functions is... more Suppressor of cytokine signaling 2 (SOCS2) is a STAT5b-regulated gene and one of its functions is to influence growth and development through negative regulatory effects on GH/IGF-1 pathway. So, we evaluate the potential influence of SOCS2 single nucleotide polymorphisms (SNPs) on clinical and laboratorial characteristics of a large cohort of Brazilian patients with acromegaly. Four SOCS2 SNPs (rs3782415, rs3816997, rs3825199 and rs11107116) were selected and genotyped by real-time PCR using specific Taqman probe assays. A total of 186 patients (116 women, age range 26-88 years) were evaluated. No association of SOCS2 genotypes was observed with none of the following clinical and laboratorial characteristics: age, sex, body mass index, comorbidities, basal GH, oral glucose tolerance test GH nadir, IGF-I, ULNR-IGF-I. Despite of the key role of SOCS2 in the regulation of GH receptor signaling, we did not find any significant association between SOCS2 polymorphisms and acromegaly.
Medical science monitor basic research, Jan 30, 2015
BACKGROUND DREAM (Downstream Regulatory Element Antagonistic Modulator) is a neuronal calcium sen... more BACKGROUND DREAM (Downstream Regulatory Element Antagonistic Modulator) is a neuronal calcium sensor that was suggested to modulate TSH receptor activity and whose overexpression provokes an enlargement of the thyroid gland in transgenic mice. The aim of this study was to investigate somatic mutations and DREAM gene expression in human multinodular goiter (MNG). MATERIAL AND METHODS DNA and RNA samples were obtained from hyperplastic thyroid glands of 60 patients (54 females) with benign MNG. DREAM mutations were evaluated by PCR and direct automatic sequencing, whereas relative quantification of mRNA was performed by real-time PCR. Over- and under-expression were defined as a 2-fold increase and decrease in comparison to normal thyroid tissue, respectively. RQ M (relative quantification mean); SD (standard deviation). RESULTS DREAM expression was detected in all nodules evaluated. DREAM mRNA was overexpressed in 31.7% of MNG (RQ M=6.26; SD=5.08), whereas 53.3% and 15% had either no...
Arquivos brasileiros de endocrinologia e metabologia
Congenital isolated hypogonadotropic hypogonadism (IHH) is characterized by partial or complete l... more Congenital isolated hypogonadotropic hypogonadism (IHH) is characterized by partial or complete lack of pubertal development due to defects in migration, synthesis, secretion or action of gonadotropin-releasing hormone (GnRH). Laboratory diagnosis is based on the presence of low levels of sex steroids, associated with low or inappropriately normal levels of pituitary gonadotropins (LH and FSH). Secretion of other pituitary hormones is normal, as well magnetic resonance imaging of the hypothalamohypophyseal tract, which shows absence of an anatomical defects. When IHH is associated with olfactory abnormalities (anosmia or hyposmia), it characterizes Kallmann syndrome. A growing list of genes is involved in the etiology of IHH, suggesting the heterogeneity and complexity of the genetic bases of this condition. Defects in olfactory and GnRH neuron migration are the etiopathogenic basis of Kallmann syndrome. Mutations in KAL1, FGFR1/FGF8, PROK2/PROKR2, NELF, CHD7, HS6ST1 and WDR11 are a...
The genetic aetiology of congenital hypopituitarism (CH) is not entirely elucidated. FGFR1 and PR... more The genetic aetiology of congenital hypopituitarism (CH) is not entirely elucidated. FGFR1 and PROKR2 loss-of-function mutations are classically involved in hypogonadotrophic hypogonadism (HH), however, due to the clinical and genetic overlap of HH and CH; these genes may also be involved in the pathogenesis of CH. Using a candidate gene approach, we screened 156 Brazilian patients with combined pituitary hormone deficiencies (CPHD) for loss-of-function mutations in FGFR1 and PROKR2. We identified three FGFR1 variants (p.Arg448Trp, p.Ser107Leu and p.Pro772Ser) in four unrelated patients (two males) and two PROKR2 variants (p.Arg85Cys and p.Arg248Glu) in two unrelated female patients. Five of the six patients harbouring the variants had a first-degree relative that was an unaffected carrier of it. Results of functional studies indicated that the new FGFR1 variant p.Arg448Trp is a loss-of-function variant, while p.Ser107Leu and p.Pro772Ser present signalling activity similar to the wi...
Abstract Background: Air pollution in industrial and urban centers is related to negative effects... more Abstract Background: Air pollution in industrial and urban centers is related to negative effects in living organisms, including alterations in male fertility. The fine inhalable particulate matter is the most deleterious fraction of air pollution and consists of a complex mixture of particles sized 2.5 µm or less of aerodynamic diameter (PM2.5). Experimental studies had already showed a deleterious effect of air pollution of Sao Paulo city on fertility in male mice. The hypothesis of its toxicity is based on activation of oxidative stress and inflammatory via as well as DNA adducts, leading to epigenetic alterations. Aim: To evaluate, under different conditions, the effects of the exposure to air pollution of city of Sao Paulo on mice testis. Methods: A group of mice was exposed during gestational period and after birth, from the weaning day until adulthood (prenatal and post-weaning group - PNPWG); another group of mice was exposed to PM2.5 during gestational period only (prenatal group - PNG); a third group of mice was exposed to PM2.5 after birth, from the weaning day until adulthood only (post-weaning group - PWG); and finally, a fourth group of mice was exposed to filtered air during gestational period and from post-weaning day until adulthood (control group - CG). Analyzes were performed on testis from adult animals. Stereological techniques were used to analyze structures of testis and Johnsen's score to evaluate spermatogenesis in a qualitative way. DNA microarray were used to evaluate gene expression. Results: Testis of all the exposed groups showed tubular degeneration characteristics. In comparison with CG, the testis of PNPWG showed higher weight and volume, surface area of the seminiferous tubules, volume of the seminiferous epithelium and epididymis weight. Quality of spermatogenesis in PNPWG was also significantly worse in comparison with CG. PWG had similar results when compared with CG, however the only parameters with significantly difference were the volume of the seminiferous epithelium and the quality of spermatogenesis. Mean volume of the seminiferous epithelium from PNG was significantly lower when compared to CG. Gene expression analysis exhibited differential expression of genes mostly related to mitochondrial tRNA in group-to-group comparison. The mt-Ty (mitochondrially encoded tRNA tyrosine) gene was hiperexpressed in PNPWG in comparison with other groups. Conclusion: We demonstrated for the first time that the exposure to PM2.5 of the city of Sao Paulo causes tubular degeneration with decreased or absent spermatogenesis associated with differential expression of genes related to mitochondrial function in mice testis. In addition, we demonstrated that post-natal exposure seems to be more deleterious for the testicular function.
A sindrome de Kallmann (SK) e uma afeccao relativamente rara, caracterizada pela associacao de hi... more A sindrome de Kallmann (SK) e uma afeccao relativamente rara, caracterizada pela associacao de hipogonadismo hipogonadotrofico (HH) e anosmia/hiposmia. Embora a maioria dos casos de SK seja esporadica, diferentes padroes de heranca, recessivo ligado ao X, autossomico dominante e autossomico recessivo tem sido descritos, sendo que a forma ligada ao X da sindrome (SKX) encontra-se genotipica e fenotipicamente melhor definida. O gene KAL-1 localizado em Xp22.3 escapa a inativacao e codifica uma proteina, a anosmina, cujo papel tem sido relacionado ao processo de migracao de neuronios produtores de GnRH que ocorre durante a embriogenese humana. No entanto, muitos casos de hipogonadismo hipogonadotrofico nao podem ser relacionados a qualquer alteracao olfativa, sendo estes classificados como hipogonadismo hipogonadotrofico normosmicos (HHn). Mutacoes no gene do receptor do hormonio liberador de gonadotrofinas (GnRH-R) tem sido descritas numa parcela desses casos. Alem disso, os genes NELF (fator nasal embrionario do hormonio liberador de hormonio luteinizante) e EBF2 (?early B-cell factor?) tem sido relacionados ao processo de migracao de neuronios produtores de GnRH, sendo considerados possivelmente envolvidos na genese do HH. O presente trabalho foi elaborado com o objetivo de se investigar a base genetica das diferentes formas do HH. Para tal, foram avaliados 31 individuos com hipogonadismo hipogonadotrofico (pertencentes a 26 familias), 22 com anosmia e nove com olfacao normal, mediante a analise do cariotipo pela tecnica de bandamento GTG seguida da analise molecular dos genes KAL-1 e GnRH-R, por PCR e sequenciamento. Todos os pacientes estudados apresentaram cariotipo normal, a excecao de um caso esporadico de SK com microdelecao do locus KAL-1 detectada somente por FISH em estudo anteriormente realizado. Entre os pacientes com SK, foram detectadas mutacoes na regiao codificante do gene KAL-1 nas tres familias com heranca recessiva ligada ao X e em um caso esporadico. Duas familias apresentaram duas delecoes intragenicas similares envolvendo os exons 5-10; na terceira familia detectou-se uma mutacao 721C>T no exon 5 levando a um codon de parada prematuro. Esta mutacao ainda nao se encontra descrita na literatura. No caso esporadico identificou-se uma mutacao ?frameshift? 1956delC no exon 12. Em relacao aos pacientes com HHn, nao foram identificadas mutacoes nos genes GnRH-R e KAL-1. Nesses casos, e nos pacientes com SK cuja etiologia nao pode ser esclarecida, foram ainda analisados os genes NELF e EBF2. Contudo, nenhuma alteracao nas sequencias desses genes foi identificada entre os pacientes da nossa casuistica. Em conclusao, nao foram encontradas evidencias cromossomicas da localizacao de outros genes candidatos responsaveis pelo surgimento do HH. Duas mutacoes novas e duas delecoes intragenicas semelhantes foram identificadas no gene KAL-1 entre os casos esporadicos e familiais da SK, observando-se que essas mutacoes ocorreram numa frequencia relativamente elevada em nossa casuistica. Os pacientes com HHn nao apresentaram mutacoes nos genes KAL-1 e GnRH-R. Nao foram observadas anomalias nas sequencias codificantes dos genes NELF e EBF2 em nenhuma das formas de HH Abstr
PurposeCushing’s disease (CD) is a severe illness generally caused by microcorticotropinomas (MIC... more PurposeCushing’s disease (CD) is a severe illness generally caused by microcorticotropinomas (MICs) and in approximately 7–20% of patients by macrocorticotropinomas (MACs). USP8-mutations have been identified as a major genetic cause of CD (~ 50%). Few studies have reported the distribution between MICs–MACs related to USP8-mutations and their genotype–phenotype correlations. Therefore, we aimed to evaluate USP8-mutations in a cohort of MICs–MACs from a unique center and to perform a systematic review and meta-analysis.MethodsDNA-tumor-tissues from 47 corticotropinomas (16 MICs and 31 MACs) were sequenced. Clinical-biochemical data, radiological imaging data and remission/recurrence rates were evaluated. In addition, we performed a meta-analysis of nine published series (n = 630).ResultsWe identified four different USP8-mutations previously described, in 11 out of 47 (23.4%) corticotropinomas; 8 out of 11 were MACs. The urinary cortisol levels of our patients with corticotrophin USP8-mutated-alleles were lower than those of patients with wild-type (WT) alleles (p ≤ 0.017). The frequency of USP8-mutated-alleles among the series was approximately 30% with a higher prevalence in female-patients (p < 0.1 × 10−4). Among the 5 series, the remission rates were higher in patients with USP8-mutated-alleles than in those with the USP8-WT-alleles (p < 0.1 × 10−4).ConclusionOur data, as well as the retrospective review of CD series associated with USP8-mutated alleles, show heterogeneous findings among the series. Several drawbacks included the lack of a systematic protocol to evaluate these patients before surgery and follow-up. Further prospective studies using a systematic protocol will provide more consistent information about the influence of the corticotropinomas with USP8-mutated alleles on the phenotype, responses to treatment and outcome of patients with CD.
Context Congenital hypogonadotropic hypogonadism (CHH) is a rare condition caused by GnRH deficie... more Context Congenital hypogonadotropic hypogonadism (CHH) is a rare condition caused by GnRH deficiency. Several genes have been associated with the pathogenesis of CHH, but most cases still remain without a molecular diagnosis. The advent of next-generation sequencing (NGS) has allowed the simultaneous genotyping of several regions, faster, making possible the extension of the genetic knowledge of CHH. Objective Genetic characterization of a large cohort of Brazilian CHH patients. Design and patients A cohort of 130 unrelated patients (91 males, 39 females) with CHH (75 normosmic CHH, 55 Kallmann syndrome) was studied using a panel containing 36 CHH-associated genes. Results Potential pathogenic or probably pathogenic variants were identified in 43 (33%) CHH patients. The genes ANOS1, FGFR1 and GNRHR were the most frequently affected. A novel homozygous splice site mutation was identified in the GNRH1 gene and a deletion of the entire coding sequence was identified in SOX10. Deleterio...
PurposeFilamin A (FLNA) expression is related to dopamine receptor type 2 (DRD2) expression in pr... more PurposeFilamin A (FLNA) expression is related to dopamine receptor type 2 (DRD2) expression in prolactinomas. Nevertheless, in corticotrophinomas, there are few studies about DRD2 expression and no data on FLNA. Therefore, we evaluated FLNA and DRD2 expression in corticotrophinomas and their association with tumor characteristics.MethodsDRD2 and FLNA expression by immunohistochemistry, using H-score, based on the percentage of positive cells in a continuous scale of 0–300, were evaluated in 23 corticotrophinomas samples from patients submitted to neurosurgery. In six patients, treatment with cabergoline was indicated after non curative surgery.ResultsTwenty-two patients were female and one male. Regarding tumor size, 10 were micro and 12 were macroadenomas. DRD2 expression was found in 89% of cases and did not correlate with FLNA expression. Moreover, the response to cabergoline, observed in 33% of the cases, did not correlate with DRD2 nor FLNA expression. FLNA expression was not associated with clinical and tumor characteristics, except for sphenoid sinus invasion.ConclusionsIn our cohort of corticotrophinomas, DRD2 expression was not associated with FLNA expression nor to the response to CAB. Nonetheless, FLNA expression could be related to tumor invasiveness.
Complete tumor removal by transsphenoidal surgery is usually difficult for large nonfunctioning p... more Complete tumor removal by transsphenoidal surgery is usually difficult for large nonfunctioning pituitary adenomas (NFPAs). A validated medical treatment may be useful for their management. This study evaluates the clinical efficacy of the dopaminergic agonist cabergoline for residual NFPA. We conducted a randomized, parallel, open-label clinical trial that compared cabergoline with nonintervention in patients with residual NFPA after transsphenoidal surgery over 2 years. The primary outcome was clinical efficacy (tumor reduction). The secondary outcome was the relationship between tumor dopamine D2 receptor (D2R) expression and clinical responsiveness. Tumor measurements and clinical evaluations were performed every 6 months. In total, 59 and 57 individuals were randomly assigned to the study and control groups, respectively. At the end of the study, residual tumor shrinkage, stabilization, and enlargement were observed in 28.8%, 66.1%, and 5.1% of patients, respectively, in the medical-therapy group and in 10.5%, 73.7%, and 15.8% of patients, respectively, in the control group (P=0.01). The progression-free survival rate was 23.2 and 20.8 months for the study and control groups, respectively (P=0.01). D2R was not associated with cabergoline responsiveness. No major side effects were related to cabergoline use. Cabergoline was an effective drug for treating residual NFPA, and its use was associated with a high rate of tumor shrinkage (ClinicalTrials.gov NCT03271918).
Tumor recurrence or incomplete resection in nonfunctioning pituitary adenomas (NFPAs) is relative... more Tumor recurrence or incomplete resection in nonfunctioning pituitary adenomas (NFPAs) is relatively common. However, predictive factors of tumor recurrence in NFPAs are not well established. We evaluated possible factors related to tumor recurrence in a large cohort of NFPAs at a single pituitary neurosurgery center. A retrospective analysis was conducted of 410 medical records of patients with NFPAs treated by transsphenoidal surgery between 2000 and 2014. Among the participants, 210 were female (51.0%). A total of 14.1% had giant adenomas. Null-cell pituitary adenomas (n = 239; 58.9%) were the most frequent, followed by silent gonadotroph adenomas (n = 112; 27.3%). Null-cell adenomas were more frequent in women (P = 0.008) and silent gonadotroph adenomas were more frequent in men (P = 0.004). Recurrence was not related to sex or age. Tumor recurrence occurred more often among silent corticotropic adenomas and giant adenomas (hazard ratio 2.45; P < 0.0001 and hazard ratio 2.35; ...
Sirtuins 1-7 (SIRT) are a highly conserved family of histone deacetylases involved in the regulat... more Sirtuins 1-7 (SIRT) are a highly conserved family of histone deacetylases involved in the regulation of longevity that have a considerable impact in transcription, DNA repair regulation, telomeric stability, cell senescence and apoptosis. In the present study, SIRT1-7 mRNA levels were evaluated in 37 somatotropinomas and 31 nonfunctioning pituitary adenomas (NFPAs) using qPCR and relation to tumor size, invasiveness and Ki-67 proliferative index was made. Overexpression of SIRT1 was observed in 86.5% of somatotropinomas versus 41.9% of NFPAs (P < 0.01). SIRT3 was more underexpressed in NFPAs than somatotropinomas (77.4 and 40.5%, respectively, P < 0.01) as well as SIRT4 and SIRT7. Despite the lack of association between sirtuins and invasiveness or Ki-67 index, SIRT1 and SIRT3 expressions were related to tumor size. Mean of the largest diameter was smaller in adenomas with SIRT1 overexpression than with normal expression (P < 0.01) and SIRT3 underexpression was associated w...
ACTH-dependent hypercortisolism caused by a pituitary adenoma [Cushing's disease (CD)] is the... more ACTH-dependent hypercortisolism caused by a pituitary adenoma [Cushing's disease (CD)] is the most common cause of endogenous Cushing's syndrome. CD is often associated with several morbidities, including hypertension, diabetes, osteoporosis/bone fractures, secondary infections, and increased cardiovascular mortality. While the majority (≈80%) of the corticotrophinomas visible on pituitary magnetic resonance imaging are microadenomas (MICs, <10 mm of diameter), some tumors are macroadenomas (MACs, ≥10 mm) with increased growth potential and invasiveness, exceptionally exhibiting malignant demeanor. In addition, larger and invasive MACs are associated with a significant increased risk of local complications, such as hypopituitarism and visual defects. Given the clinical and molecular heterogeneity of corticotrophinomas, the aim of this study was to investigate the pattern of genetic differential expression between MIC and MAC, including the invasiveness grade as a criterio...
Suppressor of cytokine signaling 2 (SOCS2) is a STAT5b-regulated gene and one of its functions is... more Suppressor of cytokine signaling 2 (SOCS2) is a STAT5b-regulated gene and one of its functions is to influence growth and development through negative regulatory effects on GH/IGF-1 pathway. So, we evaluate the potential influence of SOCS2 single nucleotide polymorphisms (SNPs) on clinical and laboratorial characteristics of a large cohort of Brazilian patients with acromegaly. Four SOCS2 SNPs (rs3782415, rs3816997, rs3825199 and rs11107116) were selected and genotyped by real-time PCR using specific Taqman probe assays. A total of 186 patients (116 women, age range 26-88 years) were evaluated. No association of SOCS2 genotypes was observed with none of the following clinical and laboratorial characteristics: age, sex, body mass index, comorbidities, basal GH, oral glucose tolerance test GH nadir, IGF-I, ULNR-IGF-I. Despite of the key role of SOCS2 in the regulation of GH receptor signaling, we did not find any significant association between SOCS2 polymorphisms and acromegaly.
Medical science monitor basic research, Jan 30, 2015
BACKGROUND DREAM (Downstream Regulatory Element Antagonistic Modulator) is a neuronal calcium sen... more BACKGROUND DREAM (Downstream Regulatory Element Antagonistic Modulator) is a neuronal calcium sensor that was suggested to modulate TSH receptor activity and whose overexpression provokes an enlargement of the thyroid gland in transgenic mice. The aim of this study was to investigate somatic mutations and DREAM gene expression in human multinodular goiter (MNG). MATERIAL AND METHODS DNA and RNA samples were obtained from hyperplastic thyroid glands of 60 patients (54 females) with benign MNG. DREAM mutations were evaluated by PCR and direct automatic sequencing, whereas relative quantification of mRNA was performed by real-time PCR. Over- and under-expression were defined as a 2-fold increase and decrease in comparison to normal thyroid tissue, respectively. RQ M (relative quantification mean); SD (standard deviation). RESULTS DREAM expression was detected in all nodules evaluated. DREAM mRNA was overexpressed in 31.7% of MNG (RQ M=6.26; SD=5.08), whereas 53.3% and 15% had either no...
Arquivos brasileiros de endocrinologia e metabologia
Congenital isolated hypogonadotropic hypogonadism (IHH) is characterized by partial or complete l... more Congenital isolated hypogonadotropic hypogonadism (IHH) is characterized by partial or complete lack of pubertal development due to defects in migration, synthesis, secretion or action of gonadotropin-releasing hormone (GnRH). Laboratory diagnosis is based on the presence of low levels of sex steroids, associated with low or inappropriately normal levels of pituitary gonadotropins (LH and FSH). Secretion of other pituitary hormones is normal, as well magnetic resonance imaging of the hypothalamohypophyseal tract, which shows absence of an anatomical defects. When IHH is associated with olfactory abnormalities (anosmia or hyposmia), it characterizes Kallmann syndrome. A growing list of genes is involved in the etiology of IHH, suggesting the heterogeneity and complexity of the genetic bases of this condition. Defects in olfactory and GnRH neuron migration are the etiopathogenic basis of Kallmann syndrome. Mutations in KAL1, FGFR1/FGF8, PROK2/PROKR2, NELF, CHD7, HS6ST1 and WDR11 are a...
The genetic aetiology of congenital hypopituitarism (CH) is not entirely elucidated. FGFR1 and PR... more The genetic aetiology of congenital hypopituitarism (CH) is not entirely elucidated. FGFR1 and PROKR2 loss-of-function mutations are classically involved in hypogonadotrophic hypogonadism (HH), however, due to the clinical and genetic overlap of HH and CH; these genes may also be involved in the pathogenesis of CH. Using a candidate gene approach, we screened 156 Brazilian patients with combined pituitary hormone deficiencies (CPHD) for loss-of-function mutations in FGFR1 and PROKR2. We identified three FGFR1 variants (p.Arg448Trp, p.Ser107Leu and p.Pro772Ser) in four unrelated patients (two males) and two PROKR2 variants (p.Arg85Cys and p.Arg248Glu) in two unrelated female patients. Five of the six patients harbouring the variants had a first-degree relative that was an unaffected carrier of it. Results of functional studies indicated that the new FGFR1 variant p.Arg448Trp is a loss-of-function variant, while p.Ser107Leu and p.Pro772Ser present signalling activity similar to the wi...
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Papers by ERICKA BARBOSA TRARBACH