RESUMEN
PURPOSE: The objectives of this study were to analyze the distribution of dual oxidase (DUOX) system genes (containing DUOX2, DUOX1, DUOXA2, and DUOXA1) variants in children with congenital hypothyroidism (CH) and their phenotypes. METHODS: Target region sequencing technology was performed on DUOX system genes among 606 CH subjects covering all the exon and intron regions. Detailed clinical data were collected for statistical analysis. RESULTS: A total of 95 suspected pathogenic variants were detected in the DUOX system genes, showing a 39.11% rate in variant carrying (237/606). DUOX2 had the highest rate in this study. There were statistical differences in maximum adjusted dose and current dose of levothyroxine between the DUOX system genes nonmutated group with the mutated group (both Psâ <â 0.001). The cases in the DUOX system genes mutated group were more likely to develop into transient CH (χâ2â =â 23.155, Pâ <â 0.001) and more likely to manifested as goiter or gland-in-situ (χâ2â =â 66.139, Pâ <â 0.001). In addition, there was no significant difference in clinical characteristics between DUOX system genes monoallelic and non-monoallelic. Although 20% of the variants affected the functional domain regions (EF hand, flavin adenine dinucleotide and nicotinamide adenine dinucleotide binding sites), there was no significant effect on the phenotype severity whether the variation is located in the functional domain regions. CONCLUSIONS: Our results showed the high variation rate of DUOX2 in the DUOX system genes among Chinese CH patients. The complex genotype-phenotype relationship of DUOX system genes broadened the understanding of CH phenotype spectrum.
Asunto(s)
Hipotiroidismo Congénito/genética , Oxidasas Duales/genética , Hipotiroidismo Congénito/enzimología , Análisis Mutacional de ADN , Femenino , Estudios de Asociación Genética , Humanos , Lactante , Recién Nacido , MasculinoRESUMEN
Weaver syndrome (WS) is a rare congenital disorder characterized by generalized overgrowth, macrocephaly, specific facial features, accelerated bone age, intellectual disability, and susceptibility to cancers. De novo mutations in the enhancer of zeste homolog 2 (EZH2) have been shown to cause WS. EZH2 is a histone methyltransferase that acts as the catalytic agent of the polycomb-repressive complex 2 (PRC2) to maintain gene repression via methylation of lysine 27 on histone H3 (H3K27). Functional studies investigating histone methyltransferase activity of mutant EZH2 from various cancers have been reported, whereas WS-associated mutations remain poorly characterized. To investigate the role of EZH2 in WS, we performed functional studies using artificially assembled PRC2 complexes containing mutagenized human EZH2 that reflected the codon changes predicted from patients with WS. We found that WS-associated amino acid alterations reduce the histone methyltransferase function of EZH2 in this in vitro assay. Our results support the hypothesis that WS is caused by constitutional mutations in EZH2 that alter the histone methyltransferase function of PRC2. However, histone methyltransferase activities of different EZH2 variants do not appear to correlate directly with the phenotypic variability between WS patients and individuals with a common c.553G>C (p.Asp185His) polymorphism in EZH2.
Asunto(s)
Anomalías Múltiples/enzimología , Anomalías Múltiples/genética , Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/genética , Anomalías Craneofaciales/enzimología , Anomalías Craneofaciales/genética , Proteína Potenciadora del Homólogo Zeste 2/genética , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Deformidades Congénitas de la Mano/enzimología , Deformidades Congénitas de la Mano/genética , N-Metiltransferasa de Histona-Lisina/genética , N-Metiltransferasa de Histona-Lisina/metabolismo , Femenino , Histona Metiltransferasas , Humanos , Lactante , Recién Nacido , Masculino , Complejo Represivo Polycomb 2/genética , Complejo Represivo Polycomb 2/metabolismoRESUMEN
BACKGROUND: Mutations in the dual oxidase maturation factor 2 (DUOXA2) and thyroid peroxidase (TPO) genes have been reported to cause goitrous congenital hypothyroidism (GCH). The aim of this study was to determine the genetic basis of GCH in affected children. METHODS: Thirty children with GCH were enrolled for molecular analysis of the DUOXA2 and TPO genes. All subjects underwent clinical examination and laboratory testing. Genomic DNA was extracted from peripheral blood leukocytes, and Sanger sequencing was used to screen for DUOXA2 and TPO gene mutations in the exon fragments amplified from the extracted DNA. Family members of those patients with mutations were also enrolled and evaluated. RESULTS: Analysis of the TPO gene revealed six genetic variants, including two novel heterozygous mutations, c.1970T> C (p.I657T) and c.2665G> T (p.G889X), and four mutations that have been reported previously (c.670_672del, c.2268dup, c.2266T> C and c.2647C> T). Three patients harbored the same mutation c.2268dup. The germline mutations from four unrelated families were consistent with an autosomal recessive inheritance pattern. Conversely, no mutations in the DUOXA2 gene were detected. CONCLUSION: Two novel inactivating mutations (c.1970T> C and c.2665G> T) in the TPO gene were identified. The c.2268dup mutation occurred frequently. No mutations in the DUOXA2 gene were detected in this study.
Asunto(s)
Autoantígenos/genética , Hipotiroidismo Congénito/genética , Yoduro Peroxidasa/genética , Proteínas de Unión a Hierro/genética , Secuencia de Bases , Estudios de Casos y Controles , Niño , Preescolar , Hipotiroidismo Congénito/enzimología , Análisis Mutacional de ADN , Femenino , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Heterocigoto , Humanos , Masculino , Proteínas de la Membrana/genética , LinajeRESUMEN
OBJECTIVES: The c.2268dup mutation in the thyroid peroxidase (TPO) gene is the most common TPO alteration reported in Taiwanese patients with thyroid dyshormonogenesis. The ancestors of these patients are believed to originate from the southern province of China. Our previous study showed that this mutation leads to reduced abundance of the TPO protein and loss of TPO enzyme activity in a Malaysian-Chinese family with goitrous hypothyroidism. The aim of our study was to provide further data on the incidence of the c.2268dup mutation in a cohort of Malaysian-Chinese and its possible phenotypic effects. SETTING: Cohort study. PARTICIPANTS: Twelve biologically unrelated Malaysian-Chinese patients with congenital hypothyroidism were recruited in this study. All patients showed high thyrotropin and low free thyroxine levels at the time of diagnosis with proven presence of a thyroid gland. PRIMARY OUTCOME MEASURE: Screening of the c.2268dup mutation in the TPO gene in all patients was carried out using a PCR-direct DNA sequencing method. SECONDARY OUTCOME MEASURE: Further screening for mutations in other exonic regions of the TPO gene was carried out if the patient was a carrier of the c.2268dup mutation. RESULTS: The c.2268dup mutation was detected in 4 of the 12 patients. Apart from the c.2268dup and a previously documented mutation (c.2647C>T), two novel TPO alterations, c.670_672del and c.1186C>T, were also detected in our patients. In silico analyses predicted that the novel alterations affect the structure/function of the TPO protein. CONCLUSIONS: The c.2268dup mutation was detected in approximately one-third of the Malaysian-Chinese patients with thyroid dyshormonogenesis. The detection of the novel c.670_672del and c.1186C>T alterations expand the mutation spectrum of TPO associated with thyroid dyshormonogenesis.
Asunto(s)
Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/genética , Predisposición Genética a la Enfermedad/genética , Bocio/genética , Yoduro Peroxidasa/genética , Mutación , Pueblo Asiatico/genética , China/etnología , Estudios de Cohortes , Hipotiroidismo Congénito/epidemiología , Exones , Femenino , Bocio/enzimología , Humanos , Lactante , Recién Nacido , Malasia/epidemiología , Masculino , Prevalencia , Análisis de Secuencia de ADN/métodosRESUMEN
Congenital hypothyroidism (CH) occurs with a prevalence of approximately 1:4000 live births. Defects of thyroid hormone synthesis account for 15-20% of these cases. Thyroid peroxidase (TPO) gene is the most common cause for dyshormonogenesis. So far, more than 60 mutations in the TPO gene have been described, resulting in a variable decrease in TPO bioactivity. We present an 8-day-old male with mild CH who was identified to have a G to A transition in the fifth codon of the TPO gene (c.13G>A; p.Ala5Thr). The unaffected family members were heterozygous carriers of the mutation, whereas 400 healthy individuals of the same ethnic background did not have the mutation. Mutation analysis of 11 known causative CH genes and 4 of our own strong candidate genes with next-generation sequencing revealed no mutations in the patient nor in any other family members. The results of in silico functional analyses indicated partial loss-of-function (LOF) in the resulting enzyme molecule due to mutation. The patient's clinical finding s were consistent with the effect of this partial LOF of the mutation. In conclusion, we strongly believe that A5T alteration in the TPO gene is actually pathogenic and suggest that it should be classified as a mutation.
Asunto(s)
Autoantígenos/genética , Hipotiroidismo Congénito/genética , Predisposición Genética a la Enfermedad/genética , Yoduro Peroxidasa/genética , Proteínas de Unión a Hierro/genética , Mutación Missense , Autoantígenos/metabolismo , Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/patología , Análisis Mutacional de ADN/métodos , Humanos , Recién Nacido , Yoduro Peroxidasa/metabolismo , Proteínas de Unión a Hierro/metabolismo , MasculinoRESUMEN
Neurodevelopment is known to be particularly susceptible to thyroid hormone insufficiency and can result in extensive structural and functional deficits within the central nervous system (CNS), subsequently leading to the establishment of cognitive impairment and neuropsychiatric symptomatology. The current study evaluated the effects of gestational and/or lactational maternal exposure to propylthiouracil (PTU)-induced hypothyroidism (as a suggestive multilevel experimental approach to the study of hypothyroidism-induced changes that has been developed and characterized by the authors) on crucial brain enzyme activities of 21-day-old Wistar rat offspring in a CNS region-specific manner. The activities of acetylcholinesterase (AChE), Na(+),K(+)-ATPase and Mg(2+)-ATPase in the offspring hypothalamus, cerebellum and pons were assessed. The study demonstrated that maternal exposure to PTU (0.05% w/v in the drinking water) during the critical periods of neurodevelopment can result in an inhibition of hypothalamic, pontine and cerebellar Na(+),K(+)-ATPase; a major marker of neuronal excitability and metabolic energy production as well as an important regulator of important systems of neurotransmission. On the other hand, no significant changes in the activities of the herein offspring CNS regions' AChE and Mg(2+)-ATPase were recorded. The observed Na(+),K(+)-ATPase inhibition: (i) is region-specific (and non-detectable in whole brain homogenetes), (ii) could constitute a central event in the pathophysiology of clinically-relevant hypothyroidism-associated developmental neurotoxicity, (iii) occurs under all examined experimental schemes, and (iv) certainly deserves further clarification at a molecular and histopathological level. As these findings are analyzed and compared to the available literature, they also underline the need for the adoption and further study of Na(+),K(+)-ATPase activity as a consistent neurochemical marker within the context of a systematic comparative study of existing (and novel) simulation approaches to congenital and early age hypothyroidism.
Asunto(s)
Encéfalo/enzimología , Hipotiroidismo/complicaciones , Complicaciones del Embarazo/enzimología , Efectos Tardíos de la Exposición Prenatal , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Acetilcolinesterasa/metabolismo , Animales , ATPasa de Ca(2+) y Mg(2+)/metabolismo , Cerebelo/enzimología , Hipotiroidismo Congénito/enzimología , Femenino , Hipotálamo/enzimología , Hipotiroidismo/inducido químicamente , Lactancia , Masculino , Puente/enzimología , Embarazo , Propiltiouracilo/administración & dosificación , Ratas , Ratas Wistar , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidoresRESUMEN
OBJECTIVE: Congenital hypothyroidism (CH) is the most common neonatal endocrine disorder and mutations in the TPO gene have been reported to cause CH. Our aim in this study was to determine the genetic basis of CH in two affected individuals coming from a consanguineous family. METHODS: Since CH is usually inherited in autosomal recessive manner in consanguineous/multi-case families, we adopted a two-stage strategy of genetic linkage studies and targeted sequencing of the candidate genes. First, we investigated the potential genetic linkage of the family to any known CH locus using microsatellite markers and then screened for mutations in linked-gene by Sanger sequencing. RESULTS: The family showed potential linkage to the TPO gene and we detected a deletion (c.2422delT) in both cases. The mutation segregated with disease status in the family. CONCLUSION: This study demonstrates that a single base deletion in the carboxyl-terminal coding region of the TPO gene could cause CH and helps to establish a genotype/phenotype correlation associated with the mutation. The study also highlights the importance of molecular genetic studies in the definitive diagnosis and accurate classification of CH.
Asunto(s)
Autoantígenos/genética , Hipotiroidismo Congénito/genética , Yoduro Peroxidasa/genética , Proteínas de Unión a Hierro/genética , Mutación , Niño , Hipotiroidismo Congénito/diagnóstico , Hipotiroidismo Congénito/enzimología , Análisis Mutacional de ADN , Femenino , Estudios de Asociación Genética , Ligamiento Genético , Predisposición Genética a la Enfermedad , Haplotipos , Herencia , Homocigoto , Humanos , Linaje , Fenotipo , Factores de Riesgo , Índice de Severidad de la Enfermedad , HermanosRESUMEN
The c.2268dup mutation in thyroid peroxidase (TPO) gene was reported to be a founder mutation in Taiwanese patients with dyshormonogenetic congenital hypothyroidism (CH). The functional impact of the mutation is not well documented. In this study, homozygous c.2268dup mutation was detected in two Malaysian-Chinese sisters with goitrous CH. Normal and alternatively spliced TPO mRNA transcripts were present in thyroid tissues of the two sisters. The abnormal transcript contained 34 nucleotides originating from intron 12. The c.2268dup is predicted to generate a premature termination codon (PTC) at position 757 (p.Glu757X). Instead of restoring the normal reading frame, the alternatively spliced transcript has led to another stop codon at position 740 (p.Asp739ValfsX740). The two PTCs are located at 116 and 201 nucleotides upstream of the exons 13/14 junction fulfilling the requirement for a nonsense-mediated mRNA decay (NMD). Quantitative RT-PCR revealed an abundance of unidentified transcripts believed to be associated with the NMD. TPO enzyme activity was not detected in both patients, even though a faint TPO band of about 80 kD was present. In conclusion, the c.2268dup mutation leads to the formation of normal and alternatively spliced TPO mRNA transcripts with a consequential loss of TPO enzymatic activity in Malaysian-Chinese patients with goitrous CH.
Asunto(s)
Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/genética , Predisposición Genética a la Enfermedad/genética , Bocio/enzimología , Bocio/genética , Yoduro Peroxidasa/genética , Polimorfismo de Nucleótido Simple/genética , China , Hipotiroidismo Congénito/diagnóstico , Femenino , Marcadores Genéticos/genética , Bocio/diagnóstico , Humanos , Recién Nacido , Malasia , Mutación/genéticaRESUMEN
BACKGROUND: Defects in the thyroid peroxidase (TPO) gene have been associated with goitrous congenital hypothyroidism (CH). CASE REPORT: In this study, we report 3 siblings possessing a homozygous mutation, c.1159G>A, but exhibiting different clinical phenotypes in a Malaysian-Malay family. The index patient was diagnosed with CH during a routine neonatal screening but the other 2 siblings appeared to be asymptomatic until the ages of 19 and 12.5, respectively, when they started to develop goiter. RESULTS AND CONCLUSION: The mutation was predicted to interrupt the correct splicing of pre-mRNA and also lead to structural alterations in the functional sites of the mutant TPO. The current results suggest the association of goiter development with a homozygous c.1159G>A mutation, but the CH in the index patient could be triggered by other genetic and epigenetic factors distinct from the c.1159G>A mutation.
Asunto(s)
Hipotiroidismo Congénito , Bocio , Homocigoto , Yoduro Peroxidasa , Mutación Puntual , Adolescente , Niño , Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/genética , Hipotiroidismo Congénito/patología , Familia , Femenino , Bocio/enzimología , Bocio/genética , Bocio/patología , Humanos , Yoduro Peroxidasa/genética , Yoduro Peroxidasa/metabolismo , Masculino , Empalme del ARN/genéticaRESUMEN
Congenital hypothyroidism is associated with delay in cell migration and proliferation in brain tissue, impairment of synapse formation, misregulation of neurotransmitters, hypomyelination and mental retardation. However, the mechanisms underlying the neuropsychological deficits observed in congenital hypothyroidism are not completely understood. In the present study we proposed a mechanism by which hypothyroidism leads to hippocampal neurotoxicity. Congenital hypothyroidism induces c-Jun-N-terminal kinase (JNK) pathway activation leading to hyperphosphorylation of the glial fibrillary acidic protein (GFAP), vimentin and neurofilament subunits from hippocampal astrocytes and neurons, respectively. Moreover, hyperphosphorylation of the cytoskeletal proteins was not reversed by T3 and poorly reversed by T4. In addition, congenital hypothyroidism is associated with downregulation of astrocyte glutamate transporters (GLAST and GLT-1) leading to decreased glutamate uptake and subsequent influx of Ca(2+) through N-methyl-D-aspartate (NMDA) receptors. The Na(+)-coupled (14)C-α-methyl-amino-isobutyric acid ((14)C-MeAIB) accumulation into hippocampal cells also might cause an increase in the intracellular Ca(2+) concentration by opening voltage-dependent calcium channels (VDCC). The excessive influx of Ca(2+) through NMDA receptors and VDCCs might lead to an overload of Ca(2+) within the cells, which set off glutamate excitotoxicity and oxidative stress. The inhibited acetylcholinesterase (AChE) activity might also induce Ca(2+) influx. The inhibited glucose-6-phosphate dehydrogenase (G6PD) and gamma-glutamyl transferase (GGT) activities, associated with altered glutamate and neutral amino acids uptake could somehow affect the GSH turnover, the antioxidant defense system, as well as the glutamate-glutamine cycle. Reduced levels of S100B and glial fibrillary acidic protein (GFAP) take part of the hypothyroid condition, suggesting a compromised astroglial/neuronal neurometabolic coupling which is probably related to the neurotoxic damage in hypothyroid brain.
Asunto(s)
Acetilcolinesterasa/metabolismo , Hipotiroidismo Congénito/enzimología , Hipocampo/enzimología , Estrés Oxidativo , Animales , Astrocitos/metabolismo , Recuento de Células , Hipotiroidismo Congénito/patología , Transportador 1 de Aminoácidos Excitadores/metabolismo , Transportador 2 de Aminoácidos Excitadores/metabolismo , Proteínas Ligadas a GPI/metabolismo , Proteína Ácida Fibrilar de la Glía/metabolismo , Ácido Glutámico/metabolismo , Hipocampo/crecimiento & desarrollo , Hipocampo/patología , Masculino , Neuronas/metabolismo , Ratas , Ratas Wistar , Subunidad beta de la Proteína de Unión al Calcio S100/metabolismo , Hormonas Tiroideas/sangreRESUMEN
Congenital hypothyroidism occurs with a prevalence of approximately 1:3 500. Defects in thyroid hormone synthesis which lead to goitrous hypothyroidism account for 10-15% of these cases. Several genetic defects have been characterized and mutations in the thyroid peroxidase (TPO) gene are the most common cause for dyshormonogenesis.So far, more than 80 mutations in the TPO gene have been described, resulting in a variable decrease in TPO bioactivity. Clinically TPO defects manifest with congenital primary goitrous hypothyroidism.We here present 2 children with congenital primary hypothyroidism, who were identified to have compound heterozygous TPO mutations. They both shared the same novel mutation in the TPO gene (C756R) in exon 13. One case presented with an apparently dominant inheritance of thyroid dyshormonogenesis.
Asunto(s)
Autoantígenos/genética , Hipotiroidismo Congénito/genética , Exones , Bocio/genética , Yoduro Peroxidasa/genética , Proteínas de Unión a Hierro/genética , Mutación Missense , Adulto , Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/patología , Familia , Femenino , Alemania , Bocio/enzimología , Bocio/patología , Humanos , Recién Nacido , MasculinoRESUMEN
Congenital hypothyroidism (CH) is the most common congenital endocrine disorder, accounting for up to 1:1500 newborns per year. CH can be related to defects in either formation and migration of the thyroid gland (dysgenesis) or thyroid hormone synthesis. The pathogenesis of dysgenetic CH is still largely unknown. On the contrary, several mutations have been found in different genes involved in thyroid dyshormonogenesis (such as pendrin, thyroperoxidase-TPO, thyroglobulin). Recently, new genes involved in the etiology of dyshormonogenesis have been identified: dual oxidase 2 (DUOX2) and dual oxidase maturation factor 2 (DUOXA2). They are the principal elements generating the hydrogen peroxide needed for TPO function. Mutations in these genes have been associated to transient or permanent CH, with a high intra and interfamilial phenotypic variability. Some hypotheses have been drawn to explain the variability of the DUOX2/A2 phenotype. Among them, the existence of other H(2)0(2) generating systems, the different requirements for thyroid hormones according to age, the ethnicity, the intake of iodine. In the present paper, the genetic and clinical features of CH caused by defects in the peroxide generator system will be revised.
Asunto(s)
Hipotiroidismo Congénito/genética , Estudios de Asociación Genética , Proteínas de la Membrana/genética , NADPH Oxidasas/genética , Envejecimiento/genética , Envejecimiento/metabolismo , Hipotiroidismo Congénito/enzimología , Oxidasas Duales , Humanos , Peróxido de Hidrógeno/metabolismo , Recién Nacido , Yoduro Peroxidasa/genética , Yoduro Peroxidasa/metabolismo , Yoduros/metabolismo , Proteínas de la Membrana/metabolismo , Mutación , NADPH Oxidasas/metabolismoRESUMEN
Dual oxidases (DUOX) 1 and 2 are components of the thyroid H(2)O(2)-generating system. H(2)O(2) is used by thyroperoxidase to oxidize iodide for thyroid hormonogenesis. Mutations in the DUOX2 gene have been described in transient and permanent congenital thyroid dyshormonogenesis. We report here a novel genetic defect causing congenital hypothyroidism in a French-Canadian patient. At neonatal screening, the patient had high TSH and low total T(4) levels. (99m)Tc scan showed a normally shaped orthotopic but mildly enlarged thyroid gland, suggesting dyshormonogenesis. Thyroxine treatment was given from 1 month to 17 years, after which it was stopped for re-evaluation and the patient remained euthyroid. The transient congenital hypothyroidism phenotype prompted us to screen for mutations in DUOX2 and DUOXA2 genes using the PCR-amplified direct sequencing method. We found complete inactivation of DUOX2 caused by a partial genomic deletion of one allele inherited from the mother associated with a paternally inherited missense mutation (c.4552G>A, p.Gly1518Ser). The deleted fragment encompasses the entire COOH-terminal end which is responsible for the NADPH-oxidase activity. The Gly1518Ser DUOX2 protein is expressed at the cell surface of transfected cells albeit at low level, but it is non-functional. This study provides further evidence that the permanent or transient nature of congenital hypothyroidism is not directly related to the number of inactivated DUOX2 alleles, suggesting the existence of other pathophysiological factors.
Asunto(s)
Dominio Catalítico/genética , Hipotiroidismo Congénito/genética , Heterocigoto , Peróxido de Hidrógeno/metabolismo , Mutación Missense/genética , NADPH Oxidasas/genética , Eliminación de Secuencia/genética , Adolescente , Adulto , Secuencia de Aminoácidos , Sustitución de Aminoácidos/genética , Secuencia de Bases , Hipotiroidismo Congénito/enzimología , Análisis Mutacional de ADN , Oxidasas Duales , Femenino , Glicosilación , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , NADPH Oxidasas/química , NADPH Oxidasas/metabolismo , EmbarazoRESUMEN
Iodine is essential for the synthesis of triiodothyronine (T3) and thyroxine (T4). Iodine deficiency leads to inadequate thyroid hormone. Hypothyroidism induced by iodine deficiency during gestation and postnatal period leads to cognitive deficits in learning and memory. However, the mechanism underlying these deficits is unclear. Calcium-dependent calmodulin kinase II (CaMKII) known as a potential memory molecule regulates important neuronal functions including learning and memory. Recent studies have shown that hypothyroidism alters phosphorylation of CaMKII in hippocampus or even in sympathetic ganglia of rats. Though the entorhinal cortex (EC) is an important functional structure within the neuronal network responsible for learning and memory, little is known about the effect of hypothyroidism on phosphorylation of CaMKII in the EC. Here, we report that iodine deficiency and propylthiouracil treatment through gestation and lactation reduce phosphorylation of CaMKII in the EC of pups. The increase of calcineurin, as well as reduction of neurogranin and calmodulin, may account for the reduced phosphorylation of CaMKII induced by developmental iodine deficiency and hypothyroidism. These findings in the EC may contribute to understanding the mechanisms that underlie impairment of learning and memory induced by developmental iodine deficiency and hypothyroidism.
Asunto(s)
Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Hipotiroidismo Congénito/enzimología , Corteza Entorrinal/enzimología , Yodo/deficiencia , Aprendizaje , Proteínas del Tejido Nervioso/metabolismo , Animales , Fosforilación , Ratas , Ratas WistarRESUMEN
The growth-retarded (grt) mouse has an autosomal recessive hypothyroidism and the female shows lifelong infertility. We previously reported that these mutant phenotypes are caused by a deficiency in the enzymatic activity of tyrosylprotein sulfotransferase-2 (TPST2), and severe thyroid hypogenesis and consequent dwarfism are mainly due to the impairment of the tyrosine sulfation of thyroid-stimulating hormone receptor (TSHR) by TPST2. Although TPST2 is ubiquitously expressed and many proteins are predicted to be tyrosine sulfated and involved in many biological processes, the functional roles of tyrosine sulfation in the reproductive organs remain unclear. These findings tempted us to hypothesize two possible mechanisms underlying the infertility; a deficiency in TPST2 activity in the reproductive organs might cause the infertility in grt mice, or a significant decrease in serum thyroid hormones might impair the normal development of reproductive organs. When mutant female mice were fed a diet supplemented with sufficient thyroid powder to correct their growth retardation, the rate of copulation, pregnancy, and parturition was completely restored. Therefore, we concluded that the infertility in grt female is due to a thyroid hormone deficiency.
Asunto(s)
Hipotiroidismo Congénito/metabolismo , Infertilidad Femenina/metabolismo , Ratones/fisiología , Sulfotransferasas/deficiencia , Hormonas Tiroideas/deficiencia , Animales , Animales de Laboratorio , Animales Recién Nacidos , Western Blotting/veterinaria , Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/genética , Femenino , Histocitoquímica/veterinaria , Infertilidad Femenina/terapia , Tamaño de la Camada , Masculino , Tamaño de los Órganos/fisiología , Ovario/fisiología , Embarazo , Sulfotransferasas/metabolismo , Hormonas Tiroideas/uso terapéutico , Útero/fisiologíaRESUMEN
CONTEXT: The recent cloning of the human iodotyrosine deiodinase (IYD) gene enables the investigation of iodotyrosine dehalogenase deficiency, a form a primary hypothyroidism resulting from iodine wasting, at the molecular level. OBJECTIVE: In the current study, we identify the genetic basis of dehalogenase deficiency in a consanguineous family. RESULTS: Using HPLC tandem mass spectrometry, we developed a rapid, selective, and sensitive assay to detect 3-monoiodo-l-tyrosine and 3,5-diodo-l-tyrosine in urine and cell culture medium. Two subjects from a presumed dehalogenase-deficient family showed elevated urinary 3-monoiodo-l-tyrosine and 3,5-diodo-l-tyrosine levels compared with 57 normal subjects without thyroid disease. Subsequent analysis of IYD revealed a homozygous missense mutation in exon 4 (c.658G>A p.Ala220Thr) that co-segregates with the clinical phenotype in the family. Functional characterization of the mutant iodotyrosine dehalogenase protein showed that the mutation completely abolishes dehalogenase enzymatic activity. One of the heterozygous carriers for the inactivating mutation recently presented with overt hypothyroidism indicating dominant inheritance with incomplete penetration. Screening of 100 control alleles identified one allele positive for this mutation, suggesting that the c.658G>A nucleotide substitution might be a functional single nucleotide polymorphism. CONCLUSIONS: This study describes a functional mutation within IYD, demonstrating the molecular basis of the iodine wasting form of congenital hypothyroidism. This familial genetic defect shows a dominant pattern of inheritance with incomplete penetration.
Asunto(s)
Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/genética , Hidrolasas/deficiencia , Hidrolasas/genética , Proteínas de la Membrana/deficiencia , Proteínas de la Membrana/genética , Adolescente , Adulto , Secuencia de Aminoácidos , Calibración , Línea Celular , Cromatografía Líquida de Alta Presión , Análisis Mutacional de ADN , Diyodotirosina/metabolismo , Diyodotirosina/orina , Femenino , Bocio/genética , Humanos , Masculino , Datos de Secuencia Molecular , Monoyodotirosina/metabolismo , Monoyodotirosina/orina , Mutación Missense , Fenotipo , Plásmidos/genética , Estándares de Referencia , Reproducibilidad de los Resultados , Tiroglobulina/metabolismo , Hormonas Tiroideas/sangre , Transfección , Adulto JovenRESUMEN
CONTEXT: Mutations in dual oxidase (DUOX2) have been proposed as a cause of congenital hypothyroidism. Previous reports suggest that biallelic mutations of DUOX2 cause permanent congenital hypothyroidism and that monoallelic mutations cause transient congenital hypothyroidism. OBJECTIVE: To clarify the inheritance of hypothyroidism, we looked at the DUOX2 gene in patients with transient congenital hypothyroidism. DESIGN: DUOX2, thyroid peroxidase, Na+/I- symporter and dual oxidase maturation factor 2 genes were analyzed in eight patients with transient congenital hypothyroidism, using the PCR-amplified direct sequencing method. PATIENTS: The eight patients were found by a neonatal screening program. Six of these patients belonged to two independent families; the other two were unrelated. Their serum TSH values varied from 24.8-233.0 mU/liter. Six of the eight patients had a low serum freeT4 level (0.19-0.84 ng/dl). Seven of the eight patients were treated with thyroid hormone replacement therapy, which ceased to be necessary by 9 yr of age. RESULTS: Eight novel mutations were detected in the DUOX2 gene. Four patients in one family were compound heterozygous for p.L479SfsX2 and p.K628RfsX10. Two patients in a second family were compound heterozygous for p.K530X and p.[E876K;L1067S]. The two remaining unrelated patients were also compound heterozygous, for p.H678R/p.L1067S and p.A649E/p.R885Q, respectively. CONCLUSION: All eight patients had biallelic mutations in the DUOX2 gene. We find that loss of DUOX2 activity results in transient congenital hypothyroidism and that transient congenital hypothyroidism caused by DUOX2 mutations is inherited as an autosomal recessive trait.
Asunto(s)
Hipotiroidismo Congénito/genética , Mutación , NADPH Oxidasas/genética , Tamizaje Neonatal , Tiroxina/uso terapéutico , Niño , Preescolar , Hipotiroidismo Congénito/enzimología , Oxidasas Duales , Femenino , Genes Recesivos , Humanos , Recién Nacido , Yoduro Peroxidasa/genética , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Simportadores/genética , Tirotropina/sangre , Tiroxina/sangre , Triyodotironina/sangreRESUMEN
AIMS: To describe the phenotype of a large group of children with congenital hypothyroidism (CH) and iodide organification defect (IOD), suspected based on normal thyroid position and abnormal perchlorate discharge test, as first step of a project evaluating correlations between phenotypes and genotypes. METHODS: 71 children born in Paris between 1980 and 2006 were included. Two groups were defined according to perchlorate discharge: total IOD (TIOD) when the release was above 90% and partial IOD (PIOD) between 10 and 90%. Comparisons between groups were performed using SPSS 14.0 for Windows. RESULTS: The incidence of IOD over the 2003-2006 period was 1:20,660. Of the 71 children, 61 had PIOD and 10 TIOD. Compared to PIOD, TIOD was characterized by greater clinical severity. A wide spectrum of clinical features was seen in the PIOD group. Evolution showed transient hypothyroidism in 10/61 patients with PIOD and 1/10 TIOD patients. CONCLUSIONS: Severe presentation in the majority of TIOD patients suggests dysfunction of a key iodide-organification enzyme. In contrast, the variety of clinical features in PIOD group suggests that diverse mechanisms may lead to PIOD, such as delayed or reduced activity of enzymes involved in hormonogenesis or defects in iodine storage and release.
Asunto(s)
Hipotiroidismo Congénito/enzimología , Yoduros/metabolismo , Glándula Tiroides/enzimología , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Paris , Estudios RetrospectivosRESUMEN
Understanding of how maternal thyroid inadequacy during early gestation poses a risk for developmental outcomes is still a challenge for the neuroendocrine community. Early neocortical neurogenesis is accompanied by maternal thyroid hormone (TH) transfer to fetal brain, appearance of TH receptors, and absence of antineurogenesis signals, followed by optimization of neuronal numbers through apoptosis. However, the effects of TH deprivation on neurogenesis and neuronal cell death before the onset of fetal thyroid are still not clear. We show that maternal TH deficiency during early gestational period causes massive premature elevation in the expression of neuronal nitric oxide synthase (nNOS) with an associated neuronal death in embryonic rat neocortex. Maternal hypothyroidism was induced by feeding methimazole (0.025% wt/vol) in the drinking water to pregnant Sprague Dawley rats from embryonic d 6. Cerebral cortices from fetuses were harvested at different embryonic stages (embryonic d 14, 16, and 18) of hypothyroid and euthyroid groups. Immunoblotting and real-time PCR results showed that both protein and RNA levels of nNOS were prematurely increased under maternal hypothyroidism, and showed reversibility upon T4 administration. Immunohistochemistry revealed an increased nNOS immunoreactivity in both the cortical plate and proliferative zone of neocortex along with a corroborative decrease in the microtubule associated protein-2 positive neurons under maternal TH insufficiency. Results combined, put forth nNOS as a novel target of maternal TH action in embryonic neocortex, and underscore the importance of prenatal screening and timely rectification of maternal TH insufficiency, even of a moderate degree.
Asunto(s)
Relaciones Materno-Fetales , Neocórtex/efectos de los fármacos , Neocórtex/embriología , Óxido Nítrico Sintasa de Tipo I/antagonistas & inhibidores , Hormonas Tiroideas/farmacología , Animales , Muerte Celular/efectos de los fármacos , Hipotiroidismo Congénito/enzimología , Hipotiroidismo Congénito/genética , Hipotiroidismo Congénito/metabolismo , Inhibidores Enzimáticos/sangre , Inhibidores Enzimáticos/farmacología , Femenino , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Neocórtex/enzimología , Neocórtex/metabolismo , Óxido Nítrico Sintasa de Tipo I/genética , Óxido Nítrico Sintasa de Tipo I/metabolismo , Fosfoproteínas Fosfatasas/metabolismo , Embarazo , Ratas , Ratas Sprague-Dawley , Hormonas Tiroideas/sangreRESUMEN
CONTEXT AND OBJECTIVE: Most cases of goitrous congenital hypothyroidism (CH) from thyroid dyshormonogenesis 1) follow a recessive mode of inheritance and 2) are due to mutations in the thyroid peroxidase gene (TPO). We report the genetic mechanism underlying the apparently dominant inheritance of goitrous CH in a nonconsanguineous family of French Canadian origin. DESIGN, SETTING, AND PARTICIPANTS: Two brothers identified by newborn TSH screening had severe hypothyroidism and a goiter with increased (99m)Tc uptake. The mother was euthyroid, but the father and two paternal uncles had also been diagnosed with goitrous CH. After having excluded PAX8 gene mutations, we hypothesized that the underlying defect could be TPO mutations. RESULTS: Both compound heterozygous siblings had inherited a mutant TPO allele carried by their mother (c.1496delC; p.Pro499Argfs2X), and from their father, one brother had inherited a missense mutation (c.1978C-->G; p.Gln660Glu) and the other an insertion (c.1955insT; p.Phe653Valfs15X). The thyroid gland of one uncle who is a compound heterozygote for TPO mutations (p.Phe653Valfs15X/p.Gln660Glu) was removed because of concurrent multiple endocrine neoplasia type 2A. Immunohistochemistry revealed normal TPO staining, implying that Gln660Glu TPO is expressed properly. Modeling of this mutant in silico suggests that its three-dimensional structure is conserved, whereas the electrostatic binding energy between the Gln660Glu TPO and its heme group becomes repulsive. CONCLUSION: We report a pedigree presenting with pseudodominant goitrous CH due to segregation of three different TPO mutations. Although goitrous CH generally follows a recessive mode of inheritance, the high frequency of TPO mutations carriers may lead to pseudodominant inheritance.