RESUMO
INTRODUCTION: KCTD15 encodes an oligomeric BTB domain protein reported to inhibit neural crest formation through repression of Wnt/beta-catenin signalling, as well as transactivation by TFAP2. Heterozygous missense variants in the closely related paralogue KCTD1 cause scalp-ear-nipple syndrome. METHODS: Exome sequencing was performed on a two-generation family affected by a distinctive phenotype comprising a lipomatous frontonasal malformation, anosmia, cutis aplasia of the scalp and/or sparse hair, and congenital heart disease. Identification of a de novo missense substitution within KCTD15 led to targeted sequencing of DNA from a similarly affected sporadic patient, revealing a different missense mutation. Structural and biophysical analyses were performed to assess the effects of both amino acid substitutions on the KCTD15 protein. RESULTS: A heterozygous c.310G>C variant encoding p.(Asp104His) within the BTB domain of KCTD15 was identified in an affected father and daughter and segregated with the phenotype. In the sporadically affected patient, a de novo heterozygous c.263G>A variant encoding p.(Gly88Asp) was present in KCTD15. Both substitutions were found to perturb the pentameric assembly of the BTB domain. A crystal structure of the BTB domain variant p.(Gly88Asp) revealed a closed hexameric assembly, whereas biophysical analyses showed that the p.(Asp104His) substitution resulted in a monomeric BTB domain likely to be partially unfolded at physiological temperatures. CONCLUSION: BTB domain substitutions in KCTD1 and KCTD15 cause clinically overlapping phenotypes involving craniofacial abnormalities and cutis aplasia. The structural analyses demonstrate that missense substitutions act through a dominant negative mechanism by disrupting the higher order structure of the KCTD15 protein complex.
Assuntos
Domínio BTB-POZ , Anormalidades Craniofaciais , Face , Humanos , Anormalidades Múltiplas , Proteínas Correpressoras/genética , Anormalidades Craniofaciais/genética , Displasia Ectodérmica , Face/anormalidades , Mutação de Sentido Incorreto/genética , SíndromeRESUMO
OBJECTIVE: Sedentary video game playing (VGP) and caloric preloads in the pre-meal environment have been shown to influence short-term food intake (FI) regulation in children. Other factors that may affect FI control include physical activity and/or heightened emotion. Therefore, we examined the effects of a glucose preload and 30 min of active VGP (aVGP) on subjective appetite, short-term FI, and subjective emotions in 9-14 year-old children. METHODS: On four test mornings approximately one-week apart, twenty-seven children (sex: 15M, 12F; age: 11.3 ± 0.3 years; BMI percentile: 55.3 ± 6.1%) consumed a standardized breakfast two hours prior to consuming 250 mL of either a 50 g glucose preload or Sucralose® control. Following the preload, participants participated in 30 min of quiet sitting or aVGP. Energy expenditure was measured during aVGP via indirect calorimetry. FI from an ad libitum pizza meal was measured after each test condition. Subjective appetite and emotions were measured at baseline (0 min), during treatment (15min), and immediately before the test meal (30 min). RESULTS: aVGP did not affect FI, but the glucose preload decreased FI compared with the sucralose control (∆ = 157 kcal, <0.001). Although not statistically significant (p=0.12), caloric compensation was lower following the glucose preload in the aVGP condition. Subjective appetite increased with time, and was higher in the sucralose control + aVGP condition (p=0.05). Change from baseline subjective emotion scores of anger and excitement were higher (p=0.03) and lower (p=0.02) after aVGP, respectively. CONCLUSIONS: Neither short-term FI nor net energy balance were affected by low-intensity aVGP (energy expenditure of 34 kcal). These findings suggest that a short bout of low-intensity aVGP does not alter energy balance during the study measurement period, and may not support achieving or maintaining healthy weights in children. However, future longitudinal studies are needed to confirm such advice.
Assuntos
Apetite , Jogos de Vídeo , Adolescente , Regulação do Apetite , Glicemia , Criança , Ingestão de Alimentos , Ingestão de Energia , Feminino , Humanos , MasculinoRESUMO
Multiple cytokines, including interleukin 6 (IL-6), IL-11, IL-27, oncostatin M (OSM), and leukemia inhibitory factor (LIF), signal via the common GP130 cytokine receptor subunit. In this study, we describe a patient with a homozygous mutation of IL6ST (encoding GP130 p.N404Y) who presented with recurrent infections, eczema, bronchiectasis, high IgE, eosinophilia, defective B cell memory, and an impaired acute-phase response, as well as skeletal abnormalities including craniosynostosis. The p.N404Y missense substitution is associated with loss of IL-6, IL-11, IL-27, and OSM signaling but a largely intact LIF response. This study identifies a novel immunodeficiency with phenotypic similarities to STAT3 hyper-IgE syndrome caused by loss of function of GP130.
Assuntos
Craniossinostoses/genética , Receptor gp130 de Citocina/genética , Síndromes de Imunodeficiência/genética , Mutação de Sentido Incorreto/genética , Pré-Escolar , Receptor gp130 de Citocina/fisiologia , Exoma/genética , Feminino , Humanos , Interleucina-11/deficiência , Interleucina-6/deficiência , Interleucinas/deficiênciaRESUMO
BACKGROUND: Craniosynostosis, the premature fusion of one or more cranial sutures, occurs in â¼1 in 2250 births, either in isolation or as part of a syndrome. Mutations in at least 57 genes have been associated with craniosynostosis, but only a minority of these are included in routine laboratory genetic testing. METHODS: We used exome or whole genome sequencing to seek a genetic cause in a cohort of 40 subjects with craniosynostosis, selected by clinical or molecular geneticists as being high-priority cases, and in whom prior clinically driven genetic testing had been negative. RESULTS: We identified likely associated mutations in 15 patients (37.5%), involving 14 different genes. All genes were mutated in single families, except for IL11RA (two families). We classified the other positive diagnoses as follows: commonly mutated craniosynostosis genes with atypical presentation (EFNB1, TWIST1); other core craniosynostosis genes (CDC45, MSX2, ZIC1); genes for which mutations are only rarely associated with craniosynostosis (FBN1, HUWE1, KRAS, STAT3); and known disease genes for which a causal relationship with craniosynostosis is currently unknown (AHDC1, NTRK2). In two further families, likely novel disease genes are currently undergoing functional validation. In 5 of the 15 positive cases, the (previously unanticipated) molecular diagnosis had immediate, actionable consequences for either genetic or medical management (mutations in EFNB1, FBN1, KRAS, NTRK2, STAT3). CONCLUSIONS: This substantial genetic heterogeneity, and the multiple actionable mutations identified, emphasises the benefits of exome/whole genome sequencing to identify causal mutations in craniosynostosis cases for which routine clinical testing has yielded negative results.
Assuntos
Craniossinostoses/genética , Genoma Humano , Sequenciamento de Nucleotídeos em Larga Escala , Proteínas de Neoplasias/genética , Craniossinostoses/diagnóstico , Craniossinostoses/patologia , Exoma/genética , Testes Genéticos , Humanos , Mutação , Valor Preditivo dos TestesRESUMO
Curry-Jones syndrome (CJS) is a multisystem disorder characterized by patchy skin lesions, polysyndactyly, diverse cerebral malformations, unicoronal craniosynostosis, iris colobomas, microphthalmia, and intestinal malrotation with myofibromas or hamartomas. Cerebellar medulloblastoma has been described in a single affected individual; in another, biopsy of skin lesions showed features of trichoblastoma. The combination of asymmetric clinical features, patchy skin manifestations, and neoplastic association previously led to the suggestion that this could be a mosaic condition, possibly involving hedgehog (Hh) signaling. Here, we show that CJS is caused by recurrent somatic mosaicism for a nonsynonymous variant in SMO (c.1234C>T [p.Leu412Phe]), encoding smoothened (SMO), a G-protein-coupled receptor that transduces Hh signaling. We identified eight mutation-positive individuals (two of whom had not been reported previously) with highly similar phenotypes and demonstrated varying amounts of the mutant allele in different tissues. We present detailed findings from brain MRI in three mutation-positive individuals. Somatic SMO mutations that result in constitutive activation have been described in several tumors, including medulloblastoma, ameloblastoma, and basal cell carcinoma. Strikingly, the most common of these mutations is the identical nonsynonymous variant encoding p.Leu412Phe. Furthermore, this substitution has been shown to activate SMO in the absence of Hh signaling, providing an explanation for tumor development in CJS. This raises therapeutic possibilities for using recently generated Hh-pathway inhibitors. In summary, our work uncovers the major genetic cause of CJS and illustrates strategies for gene discovery in the context of low-level tissue-specific somatic mosaicism.
Assuntos
Anormalidades Craniofaciais/etiologia , Intestinos/anormalidades , Mutação/genética , Anormalidades da Pele/etiologia , Receptor Smoothened/genética , Sindactilia/etiologia , Pré-Escolar , Anormalidades Craniofaciais/patologia , Feminino , Humanos , Lactente , Recém-Nascido , Intestinos/patologia , Masculino , Transdução de Sinais , Anormalidades da Pele/patologia , Sindactilia/patologiaRESUMO
Cilia are architecturally complex organelles that protrude from the cell membrane and have signalling, sensory and motility functions that are central to normal tissue development and homeostasis. There are two broad categories of cilia; motile and non-motile, or primary, cilia. The central role of primary cilia in health and disease has become prominent in the past decade with the recognition of a number of human syndromes that result from defects in the formation or function of primary cilia. This rapidly growing class of conditions, now known as ciliopathies, impact the development of a diverse range of tissues including the neural axis, craniofacial structures, skeleton, kidneys, eyes and lungs. The broad impact of cilia dysfunction on development reflects the pivotal position of the primary cilia within a signalling nexus involving a growing number of growth factor systems including Hedgehog, Pdgf, Fgf, Hippo, Notch and both canonical Wnt and planar cell polarity. We have identified a novel ENU mutant allele of Ift140, which causes a mid-gestation embryonic lethal phenotype in homozygous mutant mice. Mutant embryos exhibit a range of phenotypes including exencephaly and spina bifida, craniofacial dysmorphism, digit anomalies, cardiac anomalies and somite patterning defects. A number of these phenotypes can be attributed to alterations in Hedgehog signalling, although additional signalling systems are also likely to be involved. We also report the identification of a homozygous recessive mutation in IFT140 in a Jeune syndrome patient. This ENU-induced Jeune syndrome model will be useful in delineating the origins of dysmorphology in human ciliopathies.
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Proteínas de Transporte/genética , Comunicação Celular/genética , Cílios/patologia , Síndrome de Ellis-Van Creveld/genética , Desenvolvimento Embrionário/genética , Animais , Polaridade Celular , Cílios/genética , Modelos Animais de Doenças , Síndrome de Ellis-Van Creveld/patologia , Proteínas Hedgehog/genética , Humanos , Camundongos , Mutação , Transdução de SinaisRESUMO
Human MYO7A mutations can cause a variety of conditions involving the inner ear. These include dominant and recessive non-syndromic hearing loss and syndromic conditions such as Usher syndrome. Mouse models of deafness allow us to investigate functional pathways involved in normal and abnormal hearing processes. We present two novel mouse models with mutations in the Myo7a gene with distinct phenotypes. The mutation in Myo7a(I487N/I487N) ewaso is located within the head motor domain of Myo7a. Mice exhibit a profound hearing loss and manifest behaviour associated with a vestibular defect. A mutation located in the linker region between the coiled-coil and the first MyTH4 domains of the protein is responsible in Myo7a(F947I/F947I) dumbo. These mice show a less severe hearing loss than in Myo7a(I487N/I487N) ewaso; their hearing loss threshold is elevated at 4 weeks old, and progressively worsens with age. These mice show no obvious signs of vestibular dysfunction, although scanning electron microscopy reveals a mild phenotype in vestibular stereocilia bundles. The Myo7a(F947I/F947I) dumbo strain is therefore the first reported Myo7a mouse model without an overt vestibular phenotype; a possible model for human DFNB2 deafness. Understanding the molecular basis of these newly identified mutations will provide knowledge into the complex genetic pathways involved in the maintenance of hearing, and will provide insight into recessively inherited sensorineural hearing loss in humans.
Assuntos
Surdez/genética , Modelos Animais de Doenças , Orelha Interna/anatomia & histologia , Genes Recessivos , Sequência de Aminoácidos , Animais , Camundongos , Modelos Moleculares , Dados de Sequência Molecular , Miosina VIIa , Miosinas/química , Miosinas/genética , Homologia de Sequência de AminoácidosRESUMO
CONTEXT: The process of medical clearance screening for patients with psychiatric chief complaints has not been standardized. OBJECTIVE: To investigate the validity of a triage algorithm for psychiatric screening (TAPS) as a method to screen for the absence of acute medical illness in these patients. METHODS: The current study was a structured, retrospective medical record review in a suburban community teaching hospital with 37,000 emergency department visits per year. All ambulatory patients presenting to triage with a psychiatric chief complaint from January 31, 2001, to June 21, 2002, were assessed with TAPS. Patients with a completed TAPS and a negative assessment were identified and included in the study. A negative TAPS assessment comprised age younger than 65 years, normal vital signs, no medical complaints, no evidence of recent substance use, and no history of schizophrenia, mental retardation, or hallucinations. Emergency department records, return visit records, and inpatient admission records were reviewed for the diagnosis or management of acute medical illness. RESULTS: A total of 1179 patients were assessed with TAPS, of whom 825 (70%) had negative TAPS assessment and were eligible for inclusion. A random sample of 100 patients was selected from this group, with 7 exclusions. Sixty-six (71%) had a history of mental illness and 51 (55%) were admitted. Further, 25 (27%) had laboratory tests ordered, and none of the laboratory results required medical intervention. Twenty-nine patients (31%) received medication, mostly previously prescribed medications or sleep aids. None of the medications were for treating patients with violent or aggressive behavior. The average length of stay was 409 minutes. No patients (95% confidence interval, 0%-3%; P<.05) received a diagnosis of or treatment for acute medical illness. CONCLUSION: The TAPS form is potentially an effective tool in screening for the absence of acute medical illness.
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Algoritmos , Programas de Rastreamento/métodos , Transtornos Mentais/diagnóstico , Triagem/métodos , Doença Aguda , Adulto , Fatores Etários , Distribuição de Qui-Quadrado , Feminino , Hospitais de Ensino , Humanos , Masculino , Transtornos Mentais/psicologia , Psicometria , Estudos RetrospectivosRESUMO
BACKGROUND: Hearing impairment is the most common sensory impairment in humans, affecting 1:1,000 births. We have identified an ENU generated mouse mutant, Mozart, with recessively inherited, non-syndromic progressive hearing loss caused by a mutation in the synaptojanin 2 (Synj2), a central regulatory enzyme in the phosphoinositide-signaling cascade. METHODOLOGY/PRINCIPAL FINDINGS: The hearing loss in Mozart is caused by a p.Asn538Lys mutation in the catalytic domain of the inositol polyphosphate 5-phosphatase synaptojanin 2. Within the cochlea, Synj2 mRNA expression was detected in the inner and outer hair cells but not in the spiral ganglion. Synj2(N538K) mutant protein showed loss of lipid phosphatase activity, and was unable to degrade phosphoinositide signaling molecules. Mutant Mozart mice (Synj2(N538K/N538K)) exhibited progressive hearing loss and showed signs of hair cell degeneration as early as two weeks of age, with fusion of stereocilia followed by complete loss of hair bundles and ultimately loss of hair cells. No changes in vestibular or neurological function, or other clinical or behavioral manifestations were apparent. CONCLUSIONS/SIGNIFICANCE: Phosphoinositides are membrane associated signaling molecules that regulate many cellular processes including cell death, proliferation, actin polymerization and ion channel activity. These results reveal Synj2 as a critical regulator of hair cell survival that is essential for hair cell maintenance and hearing function.
Assuntos
Perda Auditiva Neurossensorial/enzimologia , Perda Auditiva Neurossensorial/genética , Mutagênese/genética , Mutação/genética , Proteínas do Tecido Nervoso/genética , Monoéster Fosfórico Hidrolases/genética , Sequência de Aminoácidos , Substituição de Aminoácidos/genética , Animais , Comportamento Animal , Etilnitrosoureia , Células Ciliadas Auditivas/metabolismo , Células Ciliadas Auditivas/patologia , Perda Auditiva Neurossensorial/fisiopatologia , Inositol Polifosfato 5-Fosfatases , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Dados de Sequência Molecular , Fibras Nervosas/metabolismo , Fibras Nervosas/patologia , Proteínas do Tecido Nervoso/química , Condução Nervosa/fisiologia , Monoéster Fosfórico Hidrolases/química , Gânglio Espiral da Cóclea/metabolismo , Gânglio Espiral da Cóclea/patologia , Vestíbulo do Labirinto/metabolismo , Vestíbulo do Labirinto/patologia , Vestíbulo do Labirinto/fisiopatologiaRESUMO
Double knockouts of the Msx1 and Msx2 genes in the mouse result in severe cardiac outflow tract malformations similar to those frequently found in newborn infants. Despite the known role of the Msx genes in cardiac formation little is known of the regulatory systems (ligand receptor, signal transduction and protein-DNA interactions) that regulate the tissue-specific expression of the Msx genes in mammals during the formation of the outflow tract. In the present study we have used a combination of multi-species comparative genomics, mouse transgenic analysis and in-situ hybridisation to predict and validate the existence of a remote ultra-conserved enhancer that supports the expression of the Msx1 gene in migrating mouse cardiac neural crest and the outflow tract primordia. Furthermore, culturing of embryonic explants derived from transgenic lines with agonists of the PKC and PKA signal transduction systems demonstrates that this remote enhancer is influenced by PKA but not PKC dependent gene regulatory systems. These studies demonstrate the efficacy of combining comparative genomics and transgenic analyses and provide a platform for the study of the possible roles of Msx gene mis-regulation in the aetiology of congenital heart malformation.