RESUMO
Corticotroph adenomas frequently harbor somatic USP8 mutations. These adenomas also commonly exhibit underexpression of P27, a cell cycle regulator. The present study aimed to determine the influence of USP8 mutations on clinical features of Cushing's disease and to elucidate the relationship between USP8 mutations and P27 underexpression in these tumors. Retrospective study with 32 patients with Cushing's disease was followed at the Ribeirao Preto Medical School University Hospital. We evaluated the patients' clinical data, the USP8 mutation status and the gene expression of cell cycle regulators P27/CDKN1B, CCNE1, CCND1, CDK2, CDK4, and CDK6 in tumor tissue in addition to the protein expression of P27/CDKN1B. We observed somatic mutations in the exon 14 of USP8 in 31.3% of the patients. Larger tumor size was observed in patients harboring USP8 mutations (p=0.04), with similar rates of remission, age of presentation, salivary cortisol at 23:00 h and after 1 mg dexamethasone, ACTH levels, and early postoperative plasma cortisol. We observed no differences regarding the gene or protein expression of the cell cycle regulators according to USP8 mutation status. In this Brazilian series, the observed frequency of USP8 somatic mutations was similar to that reported in European ancestry populations. Although it was reasonable that USP8 mutations could contribute to cell cycle dysregulation and P27 underexpression in corticotroph adenomas, our data did not confirm this hypothesis. It is possible that increased deubiquitinase activity observed in mutated USP8 might influence other pathways related to cell growth and proliferation.
Assuntos
Adenoma Hipofisário Secretor de ACT/genética , Ciclo Celular , Endopeptidases/genética , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Hipersecreção Hipofisária de ACTH/genética , Ubiquitina Tiolesterase/genética , Adenoma Hipofisário Secretor de ACT/fisiopatologia , Adolescente , Hormônio Adrenocorticotrópico/sangue , Adulto , Brasil , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Criança , Endopeptidases/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Éxons , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Hipersecreção Hipofisária de ACTH/metabolismo , Hipersecreção Hipofisária de ACTH/fisiopatologia , Estudos Retrospectivos , Ubiquitina Tiolesterase/metabolismo , Adulto JovemRESUMO
Familial partial lipodystrophy type 2 (FPLD2) is characterized by insulin resistance, adipose atrophy of the extremities and central obesity. Due to the resemblance with Cushing's syndrome, we hypothesized a putative role of glucocorticoid in the pathogenesis of metabolic abnormalities in FPLD2. OBJECTIVE: To evaluate the phenotypic heterogeneity and glucocorticoid sensitivity in FPLD2 patients exhibiting the p.R482W or p.R644C LMNA mutations. DESIGN, PATIENTS AND MEASUREMENTS: Prospective study with FPLD2 patients (n = 24) and controls (n = 24), who underwent anthropometric, body composition, metabolic profile and adipokines/cytokine plasma measurements. Plasma and salivary cortisol were measured in basal conditions and after 0.25, 0.5 and 1.0 mg of dexamethasone (DEX) given at 23:00 hours. Glucocorticoid receptor (GR) and 11ßHSD isoforms expression were assessed by qPCR. RESULTS: Familial partial lipodystrophy type 2 individuals presented increased waist and neck circumferences, decreased hip circumference, peripheral skinfold thickness and fat mass. Patients presented increased HOMA-IR, triglycerides, TNF-α, IL-1ß, IL-6 and IL-10, and decreased adiponectin and leptin plasma levels. FPLD2 patients showed decreased ability to suppress the HPA axis compared with controls after 0.5 mg DEX. The phenotype was more pronounced in patients harbouring the p.R482W LMNA mutation. GRß overexpression in PBMC was observed in female patients compared with female controls. CONCLUSIONS: Familial partial lipodystrophy type 2 patients exhibited anthropometric, clinical and biochemical phenotypic heterogeneity related to LMNA mutation sites and to gender. LMNA mutations affecting both lamin A and lamin C lead to more severe phenotype. FPLD2 patients also showed blunted HPA axis response to DEX, probably due to the association of increased levels of proinflammatory cytokines with GRß overexpression leading to a more severe phenotype in female.
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Glucocorticoides/farmacologia , Lipodistrofia Parcial Familiar/sangue , Lipodistrofia Parcial Familiar/metabolismo , Adiponectina/sangue , Composição Corporal/efeitos dos fármacos , Composição Corporal/fisiologia , Dexametasona/farmacologia , Feminino , Humanos , Hidrocortisona/sangue , Resistência à Insulina/genética , Interleucina-10/sangue , Interleucina-1beta/sangue , Interleucina-6/sangue , Lamina Tipo A/genética , Leptina/sangue , Lipodistrofia Parcial Familiar/genética , Masculino , Mutação/genética , Estudos Prospectivos , Isoformas de Proteínas/genética , Receptores de Glucocorticoides/genética , Fator de Necrose Tumoral alfa/sangueRESUMO
CTNNB1 mutations and abnormal ß-catenin distribution are associated with the pathogenesis of adamantinomatous craniopharyngioma (aCP). We evaluated the expression of the canonical Wnt pathway components in aCPs and its association with CTNNB1 mutations and tumor progression. Tumor samples from 14 aCP patients and normal anterior pituitary samples from eight individuals without pituitary disease were studied. Gene expression of Wnt pathway activator (WNT4), inhibitors (SFRP1, DKK3, AXIN1, and APC), transcriptional activator (TCF7), target genes (MYC, WISP2, and, CDH1), and Wnt modulator (TP53) was evaluated by qPCR. ß-Catenin, MYC, and WISP2 expression was determined by immunohistochemistry (IHC). The transcription levels of all genes studied, except APC, were higher in aCPs as compared to controls and TCF7 mRNA levels correlated with CTNNB1 mutation. CDH1 mRNA was overexpressed in tumor samples of patients with disease progression in comparison to those with stable disease. ß-Catenin was positive and aberrantly distributed in 11 out of 14 tumor samples. Stronger ß-catenin immunostaining associated positively with tumor progression. MYC positive staining was found in 10 out of 14 cases, whereas all aCPs were negative for WISP2. Wnt pathway genes were overexpressed in aCPs harboring CTNNB1 mutations and in patients with progressive disease. Recurrence was associated with stronger staining for ß-catenin. These data suggest that Wnt pathway activation contributes to the pathogenesis and prognosis of aCPs.
Assuntos
Biomarcadores Tumorais/metabolismo , Caderinas/metabolismo , Craniofaringioma/patologia , Recidiva Local de Neoplasia/patologia , Neoplasias Hipofisárias/patologia , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Adolescente , Adulto , Antígenos CD , Biomarcadores Tumorais/genética , Caderinas/genética , Estudos de Casos e Controles , Criança , Craniofaringioma/metabolismo , Craniofaringioma/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/cirurgia , Neoplasias Hipofisárias/metabolismo , Neoplasias Hipofisárias/cirurgia , Prognóstico , Proteínas Wnt/genética , Adulto Jovem , beta Catenina/genéticaRESUMO
INTRODUCTION: Studies addressing the methylation pattern in adamantinomatous craniopharyngioma (ACP) are lacking. OBJECTIVE: To identify methylation signatures in ACPs regarding clinical presentation and outcome. METHODS: Clinical and pathology data were collected from 35 ACP patients (54% male; 18.1 years [2-68]). CTNNB1 mutations and methylation profile (MethylationEPIC/Array-Illumina) were analyzed in tumoral DNA. Unsupervised machine learning analysis of this comprehensive methylome sample was achieved using hierarchical clustering and multi-dimensional scaling. Statistical associations between clusters and clinical features were achieved using Fisher's test and global biological process interpretations were aided by Gene Ontology enrichment analyses. RESULTS: Two clusters were revealed consistently by all unsupervised methods (ACP-1: n = 18; ACP-2: n = 17) with strong bootstrap statistical support. ACP-2 was enriched by CTNNB1 mutations (100% vs 56%, P = 0.0006), hypomethylated in CpG Island (CGI), non-CGI sites, and globally (P < 0.001), and associated with greater tumor size (24.1 vs 9.5cm3, P = 0.04). Enrichment analysis highlighted pathways on signaling transduction, transmembrane receptor, development of anatomical structures, cell-adhesion, cytoskeleton organization, and cytokine binding, and also cell-type specific biological processes as regulation of oligodendrocytes, keratinocyte, and epithelial cells differentiation. CONCLUSION: Two clusters of ACP patients were consistently revealed by unsupervised machine learning methods, being one of them significantly hypomethylated, enriched by CTNNB1 mutated ACPs, and associated with increased tumor size. Enrichment analysis reinforced pathways involved in tumor proliferation and in cell-specific tumoral microenvironment.
RESUMO
Children diagnosed with pediatric adrenocortical tumors (pACT) have variable outcomes, and, to date, the disease lacks robust prognostic biomarkers. The prognostic potential of tumor methylation has been demonstrated in several cancers. We aimed to evaluate the pACT methylation profile and its association with disease presentation and survival. In this cross-sectional study, we accessed the DNA methylation (MethylationEPIC Array, Illumina) of 57 primary pACT from Southeastern Brazil and the respective patients' clinicopathological features. We also applied our analysis in an independent 48 pACT methylation dataset. Unsupervised learning whole-methylome analysis showed two groups with distinct methylation signatures: pACT-1 and pACT-2. Compared to pACT-2, pACT-1 tumors were enriched with higher methylation in CpG islands, mainly in gene promoter regions. The topmost hypermethylated gene in these samples was shown to be underexpressed. Patients in the pACT-1 group were older at diagnosis and were more likely to have carcinomas and nonlocalized/advanced and recurrent/metastatic disease. Univariate and bivariate regressions showed that pACT-1 methylation signature confers superior hazard ratio of disease progression and death than known prognostic features. The methylation groups had similar frequencies of germline mutations in the TP53 gene, including the regionally frequent p.R337H. Our analysis replication validated our findings and reproduced those recently described in pACT. We demonstrated the existence of different tumor methylation signatures associated with pACT presentation and clinical evolution, even in the context of germline TP53 mutations. Our data support tumor methylation profiling as a robust and independent prognostic biomarker for pACT and suggest a list of candidate genes for further validation.
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Neoplasias do Córtex Suprarrenal , Metilação de DNA , Neoplasias do Córtex Suprarrenal/genética , Neoplasias do Córtex Suprarrenal/patologia , Biomarcadores , Biomarcadores Tumorais/genética , Criança , Ilhas de CpG , Estudos Transversais , Humanos , PrognósticoRESUMO
Objectives: To evaluate how telomere length behaves in adamantinomtous craniopharyngioma (aCP) and if it contributes to the pathogenesis of aCPs with and without CTNNB1 mutations. Design: Retrospective cross-sectional study enrolling 42 aCP patients from 2 tertiary institutions. Methods: Clinicopathological features were retrieved from the patient's charts. Fresh frozen tumors were used for RNA and DNA analyses. Telomere length was evaluated by qPCR (T/S ratio). Somatic mutations in TERT promoter (TERTp) and CTNNB1 were detected by Sanger and/or whole-exome sequencing. We performed RNA-Seq to identify differentially expressed genes in aCPs presenting with shorter or longer telomere lengths. Results: Mutations in CTNNB1 were detected in 29 (69%) tumors. There was higher frequency of CTNNB1 mutations in aCPs from patients diagnosed under the age of 15 years (85% vs 15%; P = 0.04) and a trend to recurrent disease (76% vs 24%; P = 0.1). No mutation was detected in the TERTp region. The telomeres were shorter in CTNNB1-mutated aCPs (0.441, IQR: 0.297-0.597vs 0.607, IQR: 0.445-0.778; P = 0.04), but it was neither associated with clinicopathological features nor with recurrence. RNAseq identified a total of 387 differentially expressed genes, generating two clusters, being one enriched for short telomeres and CTNNB1-mutated aCPs. Conclusions: CTNNB1: mutations are more frequent in children and adolescents and appear to associate with progressive disease. CTNNB1-mutated aCPs have shorter telomeres, demonstrating a relationship between the Wnt/ß-catenin pathway and telomere biology in the pathogenesis of aCPs.
Assuntos
Craniofaringioma , Telômero , beta Catenina , Adolescente , Criança , Craniofaringioma/genética , Estudos Transversais , Humanos , Mutação , Estudos Retrospectivos , Telômero/ultraestrutura , Via de Sinalização Wnt , beta Catenina/genéticaRESUMO
Objective: Pediatric adrenocortical tumors (pACT) display complex genomic backgrounds, lacking robust prognostic markers and targeted therapeutic options. Vitamin D3 receptor (VDR) promoter hypermethylation and underexpression were reported in adrenocortical carcinomas from adult patients. In this study, we aimed to investigate VDR expression levels and methylation status in pACT and their clinical and prognostic significance. Design: Retrospective cross-sectional study enrolling pediatric patients with ACT from two tertiary referral institutions. Methods: We evaluated clinicopathological features, VDR mRNA (qPCR) and protein (immunohistochemistry) expression, and VDR-wide methylation of ACT samples from 108 pediatric patients. Fourteen pediatric and 32 fetal and postnatal normal adrenals were used as controls. Results: Unlike in pre- and post-natal normal adrenals, most pACT lacked nuclear VDR expression and had reduced mRNA levels, especially the carcinomas. Unsupervised analysis of VDR methylation data revealed two groups of pACT with distinct disease features and outcomes. Tumors with high VDR methylation presented lower mRNA levels, and the respective patients presented advanced disease and reduced disease-free and overall survival. Conclusions: VDR has a role in normal adrenocortical development and homeostasis, which is impaired during tumorigenesis. VDR hypermethylation and underexpression may be both predictive and prognostic biomarkers for pACT.
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Neoplasias do Córtex Suprarrenal , Carcinoma Adrenocortical , Receptores de Calcitriol/metabolismo , Neoplasias do Córtex Suprarrenal/genética , Neoplasias do Córtex Suprarrenal/metabolismo , Carcinoma Adrenocortical/genética , Adulto , Biomarcadores , Criança , Estudos Transversais , Humanos , RNA Mensageiro/genética , Receptores de Calcitriol/genética , Estudos Retrospectivos , Vitamina DRESUMO
CONTEXT: The association between large for gestational age (LGA) phenotype, postnatal growth and cardiometabolic risk (CMR) in adult life remains unclear. The role of IGF1 genotype on LGA-related outcomes in adult life is unknown. AIM: To assess the postnatal growth, IGF-I levels, CMR and the influence of the 737.738 IGF1 in adults born LGA. SUBJECTS: Case-control study (n = 515) nested in a population-based prospective cohort (n = 2063); 117 LGA and 398 gender-matched controls appropriate for gestational age (AGA) subjects. METHODS: Anthropometry was evaluated at birth, at 9-10 and at 23-25 years old. At the age of 23-25 years, blood pressure (BP), glycaemia, insulinaemia, homeostasis model assessment - insulin resistance, lipids, fibrinogen, and plasma IGF-I and 737.738 IGF1 polymorphism were assessed. RESULTS: Large for gestational age subjects remained heavier and taller than AGA at 9-10 and 23-25 years (P < 0·05); at 23-25 years, LGA had greater waist circumference (WC; P < 0·05) and higher BP (P < 0·05) than controls. Body proportionality at birth did not predict metabolic outcome. LGA subjects presenting catch-down of weight in childhood had lower body mass index (BMI; P = 0·001), lower WC (P < 0·05) and lower BP (P < 0·05) at 23-25 years. 737.738 IGF-I genotype differed between groups (P < 0·001). Homozygosis for polymorphic alleles was associated with increased odds of LGA (OR: 3·2; 95% CI: 1·5-6·9), higher IGF-I (56·9 ± 16·4 vs 37·7 ± 16·0 nm; P < 0·01) and lower BP (114/68 vs 121/73 mmHg; P < 0·05). CONCLUSIONS: Young adults born LGA presented higher BMI, WC and BP and appear to be at higher CMR risk than AGA subjects. The 737.738 IGF1 polymorphism appears to play a role on birth size and LGA-related metabolic outcomes.
Assuntos
Peso ao Nascer/fisiologia , Peso Corporal/fisiologia , Coração/fisiologia , Miocárdio/metabolismo , Adulto , Antropometria/métodos , Peso ao Nascer/genética , Glicemia/metabolismo , Pressão Sanguínea/fisiologia , Peso Corporal/genética , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Criança , Feminino , Fibrinogênio/metabolismo , Frequência do Gene , Genótipo , Idade Gestacional , Humanos , Recém-Nascido , Insulina/sangue , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Lipídeos/sangue , Masculino , Polimorfismo Genético , Estudos Prospectivos , Adulto JovemRESUMO
In Cushing's syndrome, the cortisol rhythm is impaired and can be associated with the disruption in the rhythmic expression of clock genes. In this study, we evaluated the expression of CLOCK, BMAL1, CRY1, CRY2, PER1, PER2, PER3 genes in peripheral blood leukocytes of healthy individuals (n = 13) and Cushing's disease (CD) patients (n = 12). Participants underwent salivary cortisol measurement at 0900 h and 2300 h. Peripheral blood samples were obtained at 0900 h, 1300 h, 1700 h, and 2300 h for assessing clock gene expression by qPCR. Gene expression circadian variations were evaluated by the Cosinor method. In healthy controls, a circadian variation in the expression of CLOCK, BMAL1, CRY1, PER2, and PER3 was observed, whereas the expression of PER1 and CRY2 followed no specific pattern. The expression of PER2 and PER3 in healthy leukocytes presented a late afternoon acrophase, similarly to CLOCK, whereas CRY1 showed night acrophase, similarly to BMAL1. In CD patients, the circadian variation in the expression of clock genes was lost, along with the abolition of cortisol circadian rhythm. However, CRY2 exhibited a circadian variation with acrophase during the dark phase in patients. In conclusion, our data suggest that Cushing's disease, which is characterized by hypercortisolism, is associated with abnormalities in the circadian pattern of clock genes. Higher expression of CRY2 at night outlines its putative role in the cortisol circadian rhythm disruption.
Assuntos
Relógios Circadianos , Hipersecreção Hipofisária de ACTH , Proteínas CLOCK/genética , Relógios Circadianos/genética , Ritmo Circadiano , Expressão Gênica , Humanos , Hidrocortisona , LeucócitosRESUMO
CONTEXT: MicroRNAs (miRNAs) are small noncoding RNAs, functioning as antisense regulators of gene expression by targeting mRNA and contributing to cancer development and progression. More than 50% of miRNA genes are located in cancer-associated genomic regions or in fragile sites of the genome. OBJECTIVE: The aim of the study was to analyze the differential expression of let-7a, miR-15a, miR-16, miR-21, miR-141, miR-143, miR-145, and miR-150 in corticotropinomas and normal pituitary tissue and verify whether their profile of expression correlates with tumor size or remission after treatment. MATERIAL AND METHODS: ACTH-secreting pituitary tumor samples were obtained during transphenoidal surgery from patients with Cushing disease and normal pituitary tissues from autopsies. The relative expression of miRNAs was measured by real-time PCR using RNU44 and RNU49 as endogenous controls. Relative quantification of miRNA expression was calculated using the 2(-DeltaDeltaCt) method. RESULTS: We found underexpression of miR-145 (2.0-fold; P = 0.04), miR-21 (2.4-fold; P = 0.004), miR-141 (2.6-fold; P = 0.02), let-7a (3.3-fold; P = 0.003), miR-150 (3.8-fold; P = 0.04), miR-15a (4.5-fold; P = 0.03), miR-16 (5.0-fold; P = 0.004), and miR-143 (6.4-fold; P = 0.004) in ACTH-secreting pituitary tumors when compared to normal pituitary tissues. There were no differences between miRNA expression and tumor size as well as miRNA expression and ratio of remission after surgery, except in patients presenting lower miR-141 expression who showed a better chance of remission. CONCLUSION: Our results support the possibility that altered miRNA expression profile might be involved in corticotrophic tumorigenesis. However, the lack of knowledge about miRNA target genes postpones full understanding of the biological functions of down-regulated or up-regulated miRNAs in corticotropinomas.
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Adenoma Hipofisário Secretor de ACT/genética , Adenoma/genética , MicroRNAs/análise , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Hipersecreção Hipofisária de ACTH/genética , Adulto JovemRESUMO
The circadian rhythm of glucocorticoids has long been recognised within the last 75 years. Since the beginning, researchers have sought to identify basic mechanisms underlying the origin and emergence of the corticosteroid circadian rhythmicity among mammals. Accordingly, Young, Hall and Rosbash, laureates of the 2017 Nobel Prize in Physiology or Medicine, as well as Takahashi's group among others, have characterised the molecular cogwheels of the circadian system, describing interlocking transcription/translation feedback loops essential for normal circadian rhythms. Plasma glucocorticoid circadian variation depends on the expression of intrinsic clock genes within the anatomic components of the hypothalamic-pituitary-adrenal axis, which are organised in a hierarchical manner. This review presents a general overview of the glucocorticoid circadian clock mechanisms, highlighting the ontogeny of the pituitary-adrenal axis diurnal rhythmicity as well as the involvement of circadian rhythm abnormalities in the physiopathology and diagnosis of Cushing's disease.
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Relógios Circadianos , Síndrome de Cushing/diagnóstico , Glucocorticoides/metabolismo , Sistema Hipotálamo-Hipofisário/metabolismo , Sistema Hipófise-Suprarrenal/metabolismo , Ritmo Circadiano , Síndrome de Cushing/metabolismo , Síndrome de Cushing/fisiopatologia , Humanos , Sistema Hipotálamo-Hipofisário/fisiopatologia , Sistema Hipófise-Suprarrenal/fisiopatologiaRESUMO
The treatment objectives for a patient with Cushing's disease (CD) are remission of hypercortisolism, adequate management of co-morbidities, restoration of the hypothalamic-pituitary-adrenal axis, preservation of fertility and pituitary function, and improvement of visual defects in cases of macroadenomas with suprasellar extension. Transsphenoidal pituitary surgery is the main treatment option for the majority of cases, even in macroadenomas with low probability of remission. In cases of surgical failure, another subsequent pituitary surgery might be indicated in cases with persistent tumor imaging at post surgical magnetic resonance imaging (MRI) and/or pathology analysis of adrenocorticotropic hormone-positive (ACTH+) positive pituitary adenoma in the first procedure. Medical treatment, radiotherapy and adrenalectomy are the other options when transsphenoidal pituitary surgery fails. There are several options of medical treatment, although cabergoline and ketoconazole are the most commonly used alone or in combination. Novel treatments are also addressed in this review. Different therapeutic approaches are frequently needed on an individual basis, both before and, particularly, after surgery, and they should be individualized. The objective of the present review is to provide the necessary information to achieve a more effective treatment for CD. It is recommended that patients with CD be followed at tertiary care centers with experience in treating this condition.
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Hipersecreção Hipofisária de ACTH/terapia , Sociedades Médicas , Algoritmos , Brasil , HumanosRESUMO
OBJECTIVE: We aimed to determine the influence of meal time on salivary circadian cortisol rhythms and weight loss in obese women. METHODS: Twelve obese subjects (body mass index >40 kg/m(2)) were hospitalized for 64 d and then randomly assigned to one of three 18-d stages with a 5-d interval between stages. In stage 1, the subjects received a hypocaloric diet (1000 kcal/d) portioned into five meals per day. In stage 2, the subjects received the same diet between 0900 and 1100 h. In stage 3, they received the same diet between 1800 and 2000 h. Between admissions, the subjects were discharged from the hospital and consumed their usual diet at home. Salivary cortisol rhythm (in six samples collected over a 24-h period) was determined during each stage, and anthropometric, bioimpedance, indirect calorimetric, and urinary nitrogen excretion variables were measured. RESULTS: Salivary cortisol circadian rhythms were similar during all stages when measured on day 1 or 18 of treatment. Despite significant reductions in all anthropometric measurements except waist/hip ratio, no significant changes were observed in salivary cortisol rhythm after alteration of the eating hours. Starting on day 4 of treatment, nitrogen ingestion and excretion levels decreased significantly; on day 10, nitrogen balance was negative in all study stages. CONCLUSION: Administration of a hypocaloric diet led to changes in weight, body composition, resting metabolic rate, and nitrogen balance but did not significantly alter salivary circadian cortisol rhythms.
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Ritmo Circadiano/fisiologia , Ingestão de Alimentos/fisiologia , Hidrocortisona/metabolismo , Obesidade/metabolismo , Redução de Peso , Adulto , Antropometria , Metabolismo Basal/fisiologia , Composição Corporal , Calorimetria Indireta , Estudos Cross-Over , Dieta Redutora , Impedância Elétrica , Feminino , Humanos , Nitrogênio/metabolismo , Nitrogênio/urina , Obesidade/dietoterapia , Saliva/química , Redução de Peso/fisiologiaRESUMO
The postnatal synchronization of the circadian variation of the adrenal clock genes in mammals remains unknown. We evaluated the postnatal ontogeny of daily variation of clock genes (Clock/Bmal1/Per1/Per2/Per3/Cry1/Cry2/Rorα/Rev-Erbα) and steroidogenesis-related genes (Star and Mc2r) in rat adrenals and its relationship with the emergence of plasma corticosterone rhythm using cosinor analysis. Plasma corticosterone circadian rhythm was detected from postnatal day (P)1, with morning acrophase, between zeitgeber time (ZT)0 and ZT2. From P14, there was a nocturnal acrophase of corticosterone at ZT20, which was associated with pups' eye opening. From P3 there was a circadian variation of the mRNA expression of Bmal1, Per2, Per3, and Cry1 genes with morning acrophase, whereas Rev-Erbα had nocturnal acrophase. From P14, Bmal1, Per2, Per3, and Cry1 acrophases advanced by approximately 10 hours, as compared with early neonatal days, becoming vespertine-nocturnal. In all postnatal ages, Per2 and Cry1 circadian profiles were synchronized in phase with the circadian rhythm of plasma corticosterone, whereas Bmal1 was in antiphase. An adult-like Star circadian rhythm profile was observed only from P21. In conclusion, our original data demonstrated a progressive postnatal maturation of the circadian variation of the adrenal clock genes in synchrony with the development of the corticosterone circadian rhythm in rats.
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Glândulas Suprarrenais/crescimento & desenvolvimento , Glândulas Suprarrenais/metabolismo , Proteínas CLOCK/genética , Ritmo Circadiano/fisiologia , Corticosterona/metabolismo , Corticosteroides/genética , Corticosteroides/metabolismo , Animais , Animais Recém-Nascidos , Proteínas CLOCK/metabolismo , Ritmo Circadiano/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Gravidez , Ratos , Ratos WistarRESUMO
Although it is a rare condition, the accurate diagnosis and treatment of Cushing's disease is important due to its higher morbidity and mortality compared to the general population, which is attributed to cardiovascular diseases, diabetes mellitus and infections. Screening for hypercortisolism is recommended for patients who present multiple and progressive clinical signs and symptoms, especially those who are considered to be more specific to Cushing's syndrome, abnormal findings relative to age (e.g., spinal osteoporosis and high blood pressure in young patients), weight gain associated with reduced growth rate in the pediatric population and for those with adrenal incidentalomas. Routine screening is not recommended for other groups of patients, such as those with obesity or diabetes mellitus. Magnetic resonance imaging (MRI) of the pituitary, the corticotropin-releasing hormone (CRH) test and the high-dose dexamethasone suppression test are the main tests for the differential diagnosis of ACTH-dependent Cushing's syndrome. Bilateral and simultaneous petrosal sinus sampling is the gold standard method and is performed when the triad of initial tests is inconclusive, doubtful or conflicting. The aim of this article is to provide information on the early detection and establishment of a proper diagnosis of Cushing's disease, recommending follow-up of these patients at experienced referral centers. Arch Endocrinol Metab. 2016;60(3):267-86.
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Adenoma Hipofisário Secretor de ACT/diagnóstico , Adenoma/diagnóstico , Consenso , Síndrome de Cushing/diagnóstico , Adenoma Hipofisário Secretor de ACT/complicações , Adenoma/complicações , Brasil , Cromatografia Líquida de Alta Pressão , Síndrome de Cushing/etiologia , Dexametasona , Diagnóstico Diferencial , Glucocorticoides , Humanos , Hidrocortisona/sangue , Imageamento por Ressonância MagnéticaRESUMO
CONTEXT: Interindividual variation and tissue specificity of glucocorticoid (GC) sensitivity may occur in healthy subjects. OBJECTIVE AND PARTICIPANTS: The objective of this study was to evaluate the GC sensitivity in 40 healthy young subjects (21 women and 19 men; 22-42 yr old). DESIGN: We measured salivary and plasma cortisol levels before and after the administration of 0.25, 0.5, and 1 mg dexamethasone (DEX), given at 2300 h. We also evaluated the pattern of DEX-mediated inhibition of concanavalin A-stimulated peripheral blood mononuclear cell proliferation using different DEX doses, the number of binding sites, and the affinity of the GC receptor (Kd). RESULTS: The increasing DEX doses resulted in a dose-dependent decrease in cortisol levels. The majority of the subjects (70%) suppressed cortisol with DEX doses lower than 0.5 mg, and two did not suppress even with 1 mg DEX. The binding capacity was 4.1 +/- 0.3 fmol/mg protein, and the Kd was 8.1 +/- 1.3 nm. Four individuals presented with elevated Kd. Peripheral blood mononuclear cell proliferation was inhibited by DEX in a dose-dependent pattern. The median IC50 value was 7.1 x 10(-7) mol/liter. We found 77.5% (31 of 40) concordance among all three tests; 29 subjects showed all parameters between the 10th and 90th percentiles (P10-P90), one above P90, and one below P10. These two subjects could be classified as more GC resistant or sensitive, respectively. No concordance between in vivo and in vitro tests in two subjects suggested a tissue-specific sensitivity. CONCLUSIONS: This is the first report that, taking advantage of three bioassays performed on the same subject, demonstrated a considerable interindividual variability and tissue-specific GC sensitivity in a young healthy population.
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Dexametasona/farmacologia , Adulto , Relação Dose-Resposta a Droga , Resistência a Medicamentos , Feminino , Humanos , Hidrocortisona/análise , Hidrocortisona/sangue , Ativação Linfocitária/efeitos dos fármacos , Masculino , Estudos Prospectivos , Receptores de Glucocorticoides/análise , Saliva/químicaRESUMO
Anxiety and salivary cortisol were measured in subjects performing simulated public speaking (SPS), a procedure that has been neurobiologically related to panic disorder. The subjects were divided into three groups: 18 symptomatic panic patients, 16 nonsymptomatic, drug-treated panic patients, and 17 healthy controls. In the experimental session, subjective anxiety (Visual Analogue Mood Scale) and the total score of the Bodily Symptom Scale (BSS) were higher in symptomatic patients than in controls, with nonsymptomatic patients in between. Measures of cortisol taken at home showed that the level was higher at 9:00 h than at 23:00 h in every group, indicating a normal circadian regulation of the hypothalamic-pituitary-adrenal (HPA) axis in panic patients. Also in every group, the level of cortisol was high at the beginning of the experimental session and decreased after 70 min. This fall parallels the decrease in anxiety and BSS ratings, and appears to reflect habituation of initial, anticipatory anxiety. Preparation and performance of speech raised anxiety and BSS scores to the initial levels, but failed to increase cortisol measured over 60 min, starting at the end of the speech. Therefore, SPS does not seem to activate the HPA axis, as reported in panic attacks.
Assuntos
Ansiedade/diagnóstico , Ansiedade/metabolismo , Hidrocortisona/análise , Transtorno de Pânico/diagnóstico , Transtorno de Pânico/metabolismo , Saliva/química , Fala , Adulto , Antidepressivos Tricíclicos/uso terapêutico , Ansiedade/tratamento farmacológico , Ritmo Circadiano , Clomipramina/uso terapêutico , Feminino , Voluntários Saudáveis , Humanos , Sistema Hipotálamo-Hipofisário/metabolismo , Masculino , Transtorno de Pânico/tratamento farmacológico , Sistema Hipófise-Suprarrenal/metabolismo , Índice de Gravidade de DoençaRESUMO
BACKGROUND & AIMS: Lesions of hypothalamus or adjacent brain structures by the craniopharyngioma (CP) and/or its treatment, as well as changes in orexigenic and anorexigenic hormones, are possible pathogenic factors for the obesity observed in CP patients. This study assessed anthropometric measurements, food intake, and biochemical markers of CP patients. METHODS: Weight, height, skinfold thicknesses, circumferences, body composition, food intake evaluation, basal glucose, lipids, insulin, ghrelin, PYY, and HOMA-IR calculation were obtained from CP children (n = 10, 4F, aged 12 ± 4.2yr) and CP adults (n = 27,13F aged 42 ± 13 yr) and from 32 gender and age matched controls. RESULTS: Overweight/obesity was observed in 51.4% of the patients at the diagnosis and increased to 86.5% at the time of the study. Obesity was more frequent in patients with grade 2 hypothalamic involvement. Most anthropometric measurements were similar in patients and controls. Caloric intake was lower in CP adults, without difference between children. Lipid intake was higher in CP patients. Carbohydrate and protein intakes were lower in CP children, with no difference between adult groups. There were no differences in micronutrients intake as well as in insulin and PYY levels, and HOMA-IR between patients and controls. HDL-c was lower in CP adults and ghrelin higher in CP children. CONCLUSIONS: There is a high rate of overweight/obesity in CP patients at the diagnosis and throughout the follow-up period. Obesity was associated with degree of hypothalamic involvement but not with caloric intake. Obesity and a lipid rich diet may have contributed to the dyslipidemia observed in CP patients.
Assuntos
Craniofaringioma/diagnóstico , Estado Nutricional , Obesidade/diagnóstico , Hormônios Peptídicos/sangue , Neoplasias Hipofisárias/diagnóstico , Adolescente , Adulto , Antropometria , Biomarcadores/sangue , Glicemia/metabolismo , Composição Corporal , Estudos de Casos e Controles , Criança , Craniofaringioma/sangue , Craniofaringioma/fisiopatologia , Craniofaringioma/cirurgia , Dislipidemias/sangue , Dislipidemias/diagnóstico , Dislipidemias/fisiopatologia , Ingestão de Alimentos , Ingestão de Energia , Feminino , Grelina/sangue , Humanos , Hipotálamo/metabolismo , Hipotálamo/fisiopatologia , Insulina/metabolismo , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Obesidade/sangue , Obesidade/fisiopatologia , Peptídeo YY/sangue , Neoplasias Hipofisárias/sangue , Neoplasias Hipofisárias/fisiopatologia , Neoplasias Hipofisárias/cirurgia , Fatores de Tempo , Adulto JovemRESUMO
High levels of triglycerides and free fatty acids have been implicated in the pathogenesis of type 2 diabetes mellitus (DM). Congenital generalized lipodystrophy (CGL) is an autosomal recessive syndrome characterized by intense whole body reduction of subcutaneous fat. Its clinical manifestations appear during the first years of life. However, DM is usually a late event. We report a patient with CGL, diagnosed at 4 months of age, who has severe hypertriglyceridemia (serum triglyceride 12.34 mmol/l and cholesterol 3.90 mmol/l), muscular hypertrophy, hepatomegaly and DM (fasting glycemia 25.9 mmol/l). Hepatic biopsy revealed steatosis and fibrosis. A modified normolipidic (composed of medium chain triglycerides) normocaloric normoproteic milky diet and insulin therapy were instituted. After 1 month treatment a reduction of serum glucose and triglyceride levels (4.13 mmol/I and 7.7 mmol/l, respectively) was noted, with later normalization, which led to the discontinuation of insulin therapy. The patient has been maintaining good control with diet alone, presenting normal serum lipid levels (triglycerides 1.07 mmol/l, total cholesterol 2.71 mmol/l) and the following glycemic profile at OGTT: 0' 4.4 mmol/l; 30' 7.0 mmol/l; 60' 3.8 mmol/l; 90' 5.3 mmol/l, and 120' 5.2 mmol/l. The disappearance of hepatic steatosis was evidenced by a biopsy obtained 1 year after the beginning of treatment. In conolusion, this report suggests that the DM occurring in CGL can be precipitated by high triglyceride levels.
Assuntos
Diabetes Mellitus Tipo 2/etiologia , Hipertrigliceridemia/complicações , Lipodistrofia/congênito , Anormalidades Múltiplas/sangue , Glicemia/metabolismo , Diabetes Mellitus Tipo 2/diagnóstico , Feminino , Teste de Tolerância a Glucose , Hormônios/sangue , Humanos , Hipertrigliceridemia/sangue , Lactente , Insulina/sangue , Lipodistrofia/sangue , Lipodistrofia/diagnóstico , Fígado/patologia , SíndromeRESUMO
INTRODUCTION: Cortisol awakening response (CAR) is a rapid increase of cortisol levels within 30-45 min after awakening. OBJECTIVE: This study evaluates CAR compared with cortisol circadian rhythm in active and in remission Cushing's disease (CD). MATERIALS AND METHODS: We evaluated healthy controls (HC, n=19), obese (OB, n=10), in remission (n=08), and active CD patients (n=10). Salivary free cortisol (SF) was determined at 0800, 1100, 1700, 2000, and 2300 h on the first day. CAR was obtained the next morning immediately upon awakening and at 15, 30, 45, and 60-min post-wake up. RESULTS: We observed differences in SF levels throughout the day in HC, OB, and in remission CD (ANOVA P=0.0001) but not in active CD (P=0.2). We demonstrated SF increment after awakening in HC, OB, and in remission CD (ANOVA P=0.007), with no effect of time on SF in active CD. The relative increment of SF obtained at the peak after awakening (CARi%) in the active CD (67±57%) was lower than in HC (154±107%), OB (240±188%), and in remission CD (186±184%) patients (P=0.009). There was a negative correlation between the SF at awakening and the CARi% in HC (r=-0.8), OB (r=-0.78), and in remission CD (r=-0.74) but not in active CD (r=-0.35; P=0.31). CONCLUSION: This study originally described a blunted CAR in active CD in contrast to its presence in HC, OB, and in remission CD. This subtle dysfunction of the hypothalamus-pituitary-adrenal axis may represent a distinct and additional physiopathological phenomenon superimposing the dysregulated cortisol circadian rhythm in this disease.