The polymorphic inheritance of DIO2 rs225014 may predict body weight variation after Graves' disease treatment

ABSTRACT Objective: We aimed to investigate the role of DIO2 polymorphisms rs225014 and rs12885300 in Graves' disease patients, mainly for controlling body weight following treatment. Subjects and methods: We genotyped 280 GD patients by the time of diagnosis and 297 healthy control individuals using a TaqMan SNP Genotyping technique. We followed up 141 patients for 18.94 ± 6.59 months after treatment. Results: There was no relationship between the investigated polymorphisms with susceptibility to GD and gain or loss of weight after GD treatment. However, the polymorphic inheritance (CC+CT genotype) of DIO2 rs225014 was associated with a lower body weight variation after GD treatment (4.26 ± 6.25 kg) when compared to wild type TT genotype (6.34 ± 7.26 kg; p = 0.0456 adjusted for the follow-up time). This data was confirmed by a multivariate analysis (p = 0.0138) along with a longer follow-up period (p = 0.0228), older age (p = 0.0306), treatment with radioiodine (p-value = 0.0080) and polymorphic inheritance of DIO2 rs12885300 (p = 0.0306). Conclusion: We suggest that DIO2 rs225014 genotyping may have an auxiliary role in predicting the post-treatment weight behavior of GD patients.

Microarray profiling and functional analysis of differentially expressed plasma exosomal circular RNAs in Graves disease

BACKGROUND: Circulating RNA (circRNA) regulates various bioactivities in cells. A better understanding of the exosomal circRNA can provide novel insights into the pathogenesis and treatment of Graves' disease (GD). We aimed to profile the differentially expressed circRNAs (DEcRs) in plasma exosomes of patients with GD and speculate and probe the functions of the DEcR by comprehensive bioinformatics analyses. METHODS: Serum exosomes were isolated from five primary GD patients and five healthy controls via ultracentrifugation. After verification with transmission electron microscopy, exosome samples were subjected to microarray profiling using human circRNA microarrays. Two up-regulated and two down-regulated DEcRs were selected for validation in plasma exosomes from 20 GD and 20 healthy control participants using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). The circRNA/microRNA/mRNA interaction network was then assembled and the analysis of the Gene Ontology and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways was utilized to predict the potential functions of the DEcR associated genes. RESULTS: There were 15 DEcRs revealed in primary GD cases. The intronic circRNA hsa_circRNA_000102 was confirmed as an up-regulated component in plasma exosomes from patients with GD. The circRNA/microRNA/mRNA interaction network unveiled the most potential targeting microRNAs of hsa_circRNA_000102 and its associated genes. The functional analyses predicted involvement of hsa_circRNA_000102 associated genes in pathways of immune system activation, such as viral infection and interferon-beta signaling. CONCLUSIONS: hsa_circRNA_000102 is a differentially up-regulated plasma exosomal circRNA in patients with GD. Our study highlights multiple pathways, particularly virus infection and interferon-beta signaling, for mediating immune activation in Graves' disease.

Cytotoxic T Lymphocyte-associated Antigen 4 Polymorphisms Correlated with Graves' Disease in Patients of Han Ethnicity in Yunnan, China / Polimorfismos del antígeno 4 del linfocito T citotóxico correlacionados con la enfermedad de Graves en pacientes de la etnia Han en Yunnan, China

ABSTRACT Objective: To investigate the correlations between polymorphisms at position 49 in exon 1 and position 318 in the promoter of the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) gene and autoimmune thyroid diseases in a Han Chinese population. Methods: Polymerase chain reaction-restriction fragment length polymorphism was utilized. The MseI and BbvI restriction endonucleases were used to detect and analyse position 49 in exon 1 and position 318 in the promoter as well as the T/C alleles of the CTLA-4 gene in peripheral blood samples from 112 patients with Graves' disease (GD), 101 with Hashimoto's thyroiditis (HT) and 100 healthy individuals. Results: At position 49 of exon 1, the frequencies of the GG genotype and the G allele in the GD group (χ2 = 12.147; p = 0.002) were statistically significantly higher than those in the control group (χ2 = 9.925; p = 0.002), while no statistically significant differences were found between the frequencies of the GG genotype and the G allele in the HT group (χ2 = 1.195; p = 0.550) and those in the control group (χ2 = 0.984; p = 0.321). No statistically significant differences in the promoter (−318) or the T/C alleles were observed among the three groups. Position 49 in the 17th codon of exon 1 of the CTLA-4 gene may be a candidate susceptibility marker in patients of Han ethnicity with GD. Conclusion: This finding helps us to better understand the genetic risks for GD and provides a direction for targeted gene therapy.

RESUMEN Objetivo: Investigar las correlaciones entre los polimorfismos en la posición 49 en el exón 1 y la posición 318 en el promotor del gen del antígeno 4 asociado al linfocito T citotóxico (CTLA-4), con las enfermedades autoinmunes de la tiroides en una población China de Han. Métodos: Se utilizó la reacción en cadena de la polimerasa-polimorfismo de la longitud de los fragmentos de restricción. Las endonucleasas de restricción de MseI y BbvI se utilizaron para detectar y analizar la posición 49 en el exón 1 y la posición 318 en el promotor, así como los alelos T/C del gen CTLA-4 en muestras de sangre periférica de 112 pacientes con enfermedad de Graves (EG), 101 con tiroiditis de Hashimoto (TH) y 100 individuos sanos. Resultados: En la posición 49 de exón 1, las frecuencias del genotipo GG y el alelo G en el grupo de EG (χ2 = 12.147; p = 0.002) fueron estadísticamente significativamente más altas que las del grupo de control (χ2 = 9.925; p = 0.002), pero no se encontraron diferencias estadísticamente significativas entre las frecuencias del genotipo GG y el alelo G en el grupo de TH (χ2 = 1.195; p = 0.550) y las del grupo de control (χ2 = 0.984; p = 0.321). No se observaron diferencias estadísticamente significativas en el promotor (−318) ni en los alelos T/C entre los tres grupos. La posición 49 en el codón17.° del exón 1 del gen CTLA-4 puede ser un marcador de susceptibilidad candidato en pacientes de la etnia Han con EG. Conclusión: Este hallazgo nos ayuda a comprender mejor los riesgos genéticos de la EG y ofrece una dirección para la terapia génica dirigida.

Association between C1q gene polymorphisms and autoimmune thyroid diseases

ABSTRACT Objective In the present study, we aimed to assess the associations of C1q gene polymorphisms with autoimmune thyroid diseases (AITD) susceptibility. Subjects and methods A set of 1,003 AITD patients (661 with Graves' disease and 342 with Hashimoto's thyroiditis) and 880 ethnically- and geographically-matched controls from Chinese Han population were included. Five common single nucleotide polymorphisms (SNPs) (rs294185, rs292001, rs682658, rs665691 and rs294179) in C1q gene locus were genotyped. Frequencies of genotypes and alleles were compared between patients and controls, and haplotype analysis was also performed. Results There was no statistically significant difference between AITD patients and controls in the frequencies of alleles of rs294185 (P = 0.41), rs292001 (P = 0.71), rs682658 (P = 0.68), rs665691 (P = 0.68) and rs294179 (P = 0.69). There was also no statistically significant difference between AITD patients and controls in the frequencies of genotypes of rs294185 (P = 0.72), rs292001 (P = 0.89), rs682658 (P = 0.83), rs665691 (P = 0.90) and rs294179 (P = 0.43). Stratified analyses showed that none of those five SNPs in C1q gene were associated with Graves' disease or Hashimoto's thyroiditis (all P values > 0.05). Haplotype analysis revealed that there were no obvious genetic associations of C1q gene polymorphisms with AITD susceptibility. Conclusions We, for the first time, identified the associations between C1q gene SNPs and AITD, and our findings suggested that five common SNPs in C1q gene were not associated with AITD susceptibility in Chinese Han population.

Doença de Graves e deficiência de IgA como manifestações da síndrome de deleção 22q11.2 / Graves disease and IgA deficiency as manifestations of 22q11.2 deletion syndrome:

A síndrome de deleção 22q11.2 (SD22q11.2) está associada à alta variabilidade fenotípica, abrangendo o espectro velocardiofacial/síndrome de DiGeorge. Manifestações autoimunes, endocrinológicas e de imunodeficiência vêm sendo relatadas associadas à síndrome. O objetivo deste estudo foi relatar um caso de SD22q11.2 associado à deficiência de IgA e à doença de Graves e rever a literatura visando verificar a frequência dessas alterações na SD22q11.2. Os distúrbios autoimunes, cada vez mais relacionadas a SD22q11.2 e novos fenótipos, vêm sendo incorporadas ao seu espectro clínico. No presente estudo, verificou-se que a doença de Graves associada à SD22q11.2 foi relatada em apenas dezesseis pacientes e quinze descritos na literatura nos últimos 13 anos. Com base na incidência e na amplitude de seu espectro de manifestações já reconhecidas, reforçaram-se os achados da literatura de que a doença de Graves deve ser incluída nas manifestações da SD22q11.2, o que nos levaria a pesquisá-la nos portadores da deleção 22q11.2.

The 22q11.2 deletion syndrome (22q11.2DS) is related to a high phenotypic variability including the velocardiofacial/DiGeorge spectrum. Autoimmune, endocrine and immunodeficiency manifestations have been reportedly associated with the syndrome. The objective of this study was to report a case of 22q11.2DS associated with IgA deficiency and Graves disease and review literature in order to verify the frequency of syndrome alterations. Autoimmune disorders have been increasingly related to 22q11.2DS, and new phenotypes are being incorporated in the clinical spectrum of this syndrome. In our study we found that Graves disease in association with 22q11.2DS was reported in only sixteen patients, and fifteen cases were described in the last 13 years. Based on the incidence and on the amplitude of this recognized spectrum, we reinforce the findings of literature that Graves disease should be included on the 22q11.2DS manifestations, which would lead us to seek it with 22q11.2 deletion patients.

Perfil genetico de risco para doença de graves e para a oftalmopatia de graves em uma população brasileira / Genetic profile of risk to graves' disease and Graves' ophthalmology in a Brazilian population

A Doença de Graves é um processo imunológico em que a combinação de fatores genéticos e ambientais é fundamental. Vários genes têm sido propostos como envolvidos no desenvolvimento da doença, particularmente os genes do sistema HLA e os polimorfismos do gene CTLA-4. Com relação à Oftalmopatia de Graves, além dos fatores genéticos, o tabagismo é conhecido como um importante fator de seu desenvolvimento. Nós previamente demonstramos que a herança de polimorfismos em genes relacionados ao metabolismo e detoxificação de xenobióticos, além de genes relacionados a apoptose celular, como TP53, tem uma importante ação na suscetibilidade para essas doenças. Nosso objetivo foi determinar as relações entre o polimorfismo do gene CTLA-4 região promotora -318, CYP1A1m1, GSTP1 e 72TP53 e os riscos para Doença de Graves e Oftalmopatia de Graves. Avaliar a relação entre fatores clínicos (idade, sexo, etnia, tabagismo, tamanho do bócio), laboratoriais (TSH, T4livre, T3total, ANTITPO, antiTg, TRAb), de imagem (captação tiroidiana com tecnécio ou iodo131) e de tipo de tratamento (radioiodo, drogas antitiroidianas e cirurgia) . Estudamos um total de 193 pacientes com Doença de Graves comparados com 200 indivíduos-controle, pareados por idade e etnia. A análise genética foi realizada através de PCR-RFLP em DNA extraído de sangue periférico. Confirmando nossos dados anteriores, as variantes dos genes GSTP1 (p = 0,0007) e CYP1A1m1 (p < 0,0001 ) eram mais frequentes em pacientes com Doença de Graves do que em controles, mas isso não ocorreu com o gene CTLA-4, região promotora -318 (p = 0,12) ou nas variantes 72TP53 (p = 0,27). Estudando o mesmo polimorfismo na Oftalmopatia de Graves, observamos que o genótipo...

Graves' disease is an immunologic process in which the combination of environmental and genetic factors is fundamental. Various genes have been proposed as involved in the development of the disease, particularly HLA system genes and the polymorphisms of CTLA-4 gene. Concerning Graves' ophthalmopathy, besides the genetic factor, smoking is a well accepted factor of its development. We previously demonstrated that the inheritance of polymorphisms in genes related to the metabolism and detoxification of xenobiotics, such as CYP1A1 and GSTP1 genes, besides the genes related to cellular apoptosis, such as TP53, have an important role in the susceptibility to these diseases. Our objective was to determine the relationship among CTLA-4 gene and CYP1A1m1, GSTM1, GSTP1 and 72TP53 genes in the risk for Graves' disease. We studied a total 193 Graves' disease patients compared to 200 control individuals, matched for age and ethnicity. The genetic analysis was done with the use of PCR-RFLP in DNA extracted from peripheral blood. Reinforcing our previous data, GSTP1 (p=0.0007) and CYP1A1m1 (p<0.0001) variants were more frequent among Graves' disease patients than in controls, but this did not happen to CTLA-4 position 318 (p=0.12) or to TP53 variants...

Fatal outcome of a young woman with papillary thyroid carcinoma and graves' disease: possible implication of "cross-signalling" mechanism / Desfecho Fatal de uma Paciente com Carcinoma Papilífero de Tiróide e Doença de Graves: possível Implicação do Mecanismo de Sinalização Cruzada

A 29 yrs-old patient was referred to our hospital due to generalized convulsions. She had hyperthyroidism treated with methimazole. Her MRI showed 4 metastatic lesions in the brain. She had a goiter with a "cold" nodule and a palpable ipsilateral lymph node. The FNAB disclosed a papillary thyroid carcinoma. Under 5 mg of MMI treatment, she had a subclinical hyperthyroidism and TRAb were 47.8 percent (n.v. < 10 percent). The CT scan also showed lung metastasis. She underwent a total thyroidectomy with a modified neck dissection and she received an accumulated radioiodine dose of 700 mCi during the following two years. She died from the consequences of multiple metastatic lesions. Studies were performed in DNA extracted from paraffin-embedded tissue from the tumor, the metastatic lymph node and the non-tumoral thyroid. The genetic analysis of tumoral DNA revealed point mutations in two different genes: the wild type CAA at codon 61 of N-RAS mutated to CAT, replacing glycine by histidine (G61H) and the normal GCC sequence at codon 623 of the TSHR gene was replaced by TCC, changing the alanine by serine (A623S). In the non-tumoral tissue no mutations were found. In vitro studies showed a constitutive activation of the TSHR. It is very probable that this activating mutation of the TSHR is unable to reach the end point of the PKA cascade in the tumoral tissue. One possibility that could explain this is the presence of a cross-signaling mechanism generating a deviation of the TSH receptor cascade to the more proliferative one involving the MAPKinase, giving perhaps a more aggressive behavior of this papillary thyroid cancer.

Paciente de 29 anos foi encaminhada ao Hospital de Clínicas por causa de convulsões generalizadas. Apresentava hipertiroidismo tratado com metimazol (MMI). A ressonância magnética mostrava quatro lesões metastáticas cerebrais. Possuía bócio com nódulo frio e linfonodo palpável ipsilateral. Usando 5 mg de MMI, a paciente apresentava hipertiroidismo subclínico e TRAb = 47,8 por cento (normal < 10 por cento). A tomografia computadorizada também mostrava metástases pulmonares. A paciente foi submetida a tiroidectomia total com dissecção cervical modificada e recebeu dose acumulada de radioiodo de 700 mCi durante o período de dois anos. Foi analisado o DNA extraído de tecido emblocado em parafina do tumor, do linfonodo metastático e de tecido tiroidiano não-tumoral. Foram encontradas mutações pontuais em dois genes: uma substituição do genótipo selvagem CAA no códon 61 de /N-RAS/ por CAT, substituindo a glicina pela histidina (G61H) e uma substituição da seqüência normal GCC no códon 623 do gene TSHR por TCC, trocando a alanina pela serina (A623S). Não foram encontradas mutações no tecido não-tumoral. Estudos in vitro mostraram ativação constitutiva de TSHR. Já que esta mutação ativadora de TSHR foi incapaz de atingir o final da cascata PKA no tecido tumoral, sugere-se que um mecanismo de cross-signaling possa explicar o desvio da cascata do receptor de TSH para outra mais proliferativa, envolvendo MAPKinase e levando ao comportamento mais agressivo deste câncer papilífero.

Association of TNF-alpha, TNF-beta, IFN-gamma and IL-1Ra gene polymorphisms with Graves' disease in the Thai population.

Cytokines play a key role in the regulation of immune and inflammatory responses. Therefore, cytokine genes are potentially related to susceptibility to Graves' disease (GD). The aim of this study was to investigate the putative functional polymorphisms within tumor necrosis factor-alpha (TNF-alpha), tumor necrosis factor-beta (TNF-beta), interferon-gamma (IFN-gamma), and interleukin-1 receptor antagonist (IL-1Ra) genes, in patients with GD (n = 137) compared to a healthy Thai control group (n = 137). The results showed no statistically significant difference between the study groups for TNF-beta (Ncol site in intron 1), IFN-gamma (+874 in intron 1), and IL-1Ra (variable numbers of tandem repeats in intron 2) gene polymorphisms. Only the -863A allele within the promoter region of the TNF-alpha gene, which may affect the affinity of the promoter nuclear factor (NF)-kappab interaction, was found to be increased in GD patients compared to the controls (p = 0.009, OR = 1.8, 95% CI = 1.15 to 2.84). The effect of the -863A allele of the TNF-alpha gene was similar to the autosomal dominance mode of inheritance (p = 0.01, OR = 2, 95% CI = 1.16 to 3.44). This polymorphism may be involved in the susceptibility to GD in part through its higher promoter activity of TNF-alpha production.

No association between an interleukin 4 gene promoter (-589) polymorphism and Graves' disease in Thai patients.

Cytokines play a key role in the regulation of immune and inflammatory responses and therefore are potential candidate genes for autoimmune thyroid disease. Polymorphisms in cytokine genes may effect gene transcription, causing individual variations in cytokine production. Several investigators have linked the interleukin-4 (IL-4) gene and autoimmune disease. The present population-based study was to investigate the polymorphisms of IL-4 gene promoter (-589C/T) in GD patients compared with a control group and determine the association with GD in a Thai population. The subjects included 137 GD patients and 137 healthy control subjects with similar ethnic and geographic backgrounds. The IL-4 gene polymorphism at position -589 in the promoter was analyzed using the PCR-RFLP. The protective effect of the -589T allele as suggested by Hunt et al in a Caucasian population was not observed in the present study. The -589T allele frequencies were similar between patients and control subjects (69% vs 69.3%) suggesting that this polymorphism can not be used as a genetic marker for GD susceptibility in Thais.

Associaçäo de alelos do complexo principal de histocompatibilidade de classe II com a Doença de Graves, em pacientes brasileiros / Association of major histocompatibility complex class II alleles with Graves' Disease in brazilian patients

A Doença de Graves (DG) é a causa mais freqüente de hipertireoidismo. Sua origem é multifatorial, complexa, na qual a susceptibilidade genética interage com o meio ambiente e fatores endógenos para causar a doença. Os alelos do Complexo Principal de Histocompatibilidade (CPH) de classe II têm sido associados com a DG, em populações de diferentes etnias e muitas evidências apontam para a associaçäo do antígeno HLA-DR3 em caucasianos. O envolvimento dos alelos do CPH na nossa populaçäo, altamente miscigenada, foi estudado em pacientes com a DG e em indivíduos-controle da mesma área geográfica, utilizando-se DNA genômico amplificado e hibridado com iniciadores de sequência específica (SSP). O alelo HLA-DRB1*0301 esteve presente com maior freqüência (44,7 por cento) em pacientes com DG, relativamente à populaçäo-controle (22,3 por cento, pc=0,0068), conferindo Risco Relativo (RR) de 2,8 e uma Fraçäo Etiológica de 28,7, enquanto o alelo HLA-DQB1*0602 esteve, significativamente, diminuído nesses pacientes (8 por cento) em relaçäo aos pacientes-controle (31,9 por cento, pc=0,00062), conferindo RR de 1,8 e uma Fraçäo Preventiva de 26,7. Apesar de a populaçäo brasileira ser altamente miscigenada, a confirmaçäo do alelo HLA-DRB1*0301, conferindo susceptibilidade à doença, aponta esse alelo com um marcador importante na predisposiçäo à doença. Em contraste, a proteçäo conferida pelo alelo HLA-DQB1*0602 parece ser peculiar aos pacientes brasileiros

Lack of association between a polymorphism of human thyrotropin receptor gene and autoimmune thyroid disease.

A polymorphism in codon 52 of the human thyrotropin receptor results in a proline to threonine substitution in the extracellular domain of the receptor, but the association with autoimmune thyroid disease has been uncertain and there is no report the prevalence of this polymorphism in Orientals. To investigate this polymorphism and the association with autoimmune thyroid disease, we studied 113 normal unrelated individuals, 142 autoimmune thyroid disease patients including 112 Graves' disease and 30 Hashimoto's thyroiditis in the Thai population. We screened genomic DNAs of these subjects for the presence of A253 by PCR amplification using a degenerate oligonucleotide primer which produces a Tth111 I restriction site only in the presence of A253. The variant allele was present in 5.3 per cent of normal and 3.5 per cent of autoimmune thyroid disease, 2.7 per cent of Graves' disease and 6.7 per cent of Hashimoto's thyroiditis. The allele distribution in autoimmune thyroid disease patients did not differ significantly from that observed in controls. No association was found between this TSH-R polymorphism and the occurrence of autoimmune thyroid disease.

Avaliaçäo fenotípica dos infiltrados celulares e da expressäo de HLA-DR em tireóides de pacientes com doença de Graves tratados com propiltiouracil / Phenotype evaluation of the cell infiltrated and of the HLA-DR expression in the thyroid of patients with Graves' disease treated with propylthiouracil

É sabido que as tireóides de pacientes com doença de Graves apresentam uma infiltraçao por mononucleares, principalmente linfócitos. Com a intençao de se avaliar estes infiltrados linfocitários, células T e a associaçao entre estes e o HLA-DR, caso os tireócitos estejam positivos para este antígeno de classe II, oito amostras de tecido tireoideano, obtidas cirurgicamente de oito pacientes com a doença de Graves e tratados com propiltiouracil (PTU), foram estudadas em secçoes criostáticas utilizando-se anticorpos monoclonais específicos (UCHT1: CD3, 16H5: CD4, OKT8: CD8, OKM1: CD14 e MID3: HLA-DR) através de imuno-histoquímica (APAAP). Em sete dos oito pacientes foi identificada a presença de células T, com predominância de células CD4+ sobre as CD8+ em quase todos. A expressao anormal de HLA-DR nos tireócitos foi observada em todos os tecidos estudados, sendo que aqueles com maior densidade de infiltraçao linfocitária foram os que apresentaram a maior positividade para o HLA-DR. Nao houve significância estatística quando testadas as combinaçoes HLA-DR e a infiltraçao linfocitária total, as subpopulaçoes linfocitárias entre si (CD4+ e CD8+) e cada uma destas isoladamente com as células T total (CD3+). Uma provável influência do PTU nos infiltrados celulares intraglandulares, como na expressao anormal de HLA-DR nos tireócitos, nao foi possível de ser avaliada, pois todos os tecidos estudados foram expostos a esta droga antitireoideana.

Estudio de familiares de pacientes con bocio tóxico difuso / Study of relatives of patients with diffuse toxic goiter

Se realizó un estudio de la función tiroidea mediante las pruebas de estimulación con hormona hipotálamica liberadora de tirotropina y de inhibición I-T3 en 100 familias de primer grado de pacientes con Bocio Tóxico Difuso (BTD) con el objetivo de detectar alteraciones preclínicas (inmunológicas y de la función tiroidea) que pudieran servir como marcadores subclínicos, conocer la repercusión hipotálamo-hipofisaria de la disfunción tiroidea subclínica y precisar características personales así como la respuesta del Eje hipotálamo-hipofisotiroideo ante el estímulo emocional. Se encontraron trastornos de la función tiroidea expresadas como una respuesta anómala de Tirotrofina a la hormona hipotálamica liberadora de Tirotrofina en el 24 % de las casos, así como no en el 7 % evolutivamente después de un período de seguimiento que osciló entre 1 y 5 años. Los cambios encontrados en la función tiroidea de estos familiares así como los reportados en pacientes con BTD sugieren una posible heterogeneidad de esta afección la que de ser demostrada implicaría la posibilidad de establecer diagnóstico precoz de esta entidad. Se encontraron características personales similares a la de los pacientes con BTD no tratados