RESUMEN
OBJECTIVE: To identify the molecular genetic aetiology of a family with autosomal dominant amelogenesis imperfecta (AI). SUBJECTS AND METHODS: DNA samples were collected from a six-generation family, and the candidate gene approach was used to screen for the enamelin (ENAM) gene. Whole-exome sequencing and linkage analysis with SNP array data identified linked regions, and candidate gene screening was performed. RESULTS: Mutational analysis revealed a mutation (c.561_562delCT and p.Tyr188Glnfs*13) in the DLX3 gene. After finding a recurrent DLX3 mutation, the clinical phenotype of the family members was re-examined. The proband's mother had pulp elongation in the third molars. The proband had not hair phenotype, but her cousin had curly hair at birth. CONCLUSIONS: In this study, we identified a recurrent 2-bp deletional DLX3 mutation in a new family. The clinical phenotype was the mildest one associated with the DLX3 mutations. These results will advance the understanding of the functional role of DLX3 in developmental processes.
Asunto(s)
Amelogénesis Imperfecta/genética , Secuencia de Bases , Proteínas de Homeodominio/genética , Eliminación de Secuencia , Factores de Transcripción/genética , Adulto , Amelogénesis Imperfecta/diagnóstico por imagen , Niño , Análisis Mutacional de ADN , Exoma , Femenino , Humanos , Masculino , Linaje , Fenotipo , Radiografía DentalRESUMEN
In order to achieve highly mineralized tooth enamel, enamel proteinases serve the important function of removing the remaining organic matrix in the mineralization and maturation of the enamel matrix. Mutations in the kallikrein 4 (KLK4), enamelysin (MMP20), and WDR72 genes have been identified as causing hypomaturation enamel defects in an autosomal-recessive hereditary pattern. In this report, 2 consanguineous families with a hypomaturation-type enamel defect were recruited, and mutational analysis was performed to determine the molecular genetic etiology of the disease. Whole exome sequencing and autozygosity mapping identified novel homozygous mutations in the KLK4 (c.620_621delCT, p.Ser207Trpfs*38) and MMP20 (c.1054G>A, p.Glu352Lys) genes. Further analysis on the effect of the mutations on the translation, secretion, and function of KLK4 and MMP20 revealed that mutant KLK4 was degraded intracellularly and became inactive while mutant MMP20 was expressed at a normal level but secreted only minimally with proteolytic function.
Asunto(s)
Amelogénesis Imperfecta/genética , Calicreínas/genética , Metaloproteinasa 20 de la Matriz/genética , Mutación , Amelogénesis Imperfecta/diagnóstico por imagen , Western Blotting , Niño , Consanguinidad , Femenino , Genotipo , Homocigoto , Humanos , Linaje , Reacción en Cadena de la Polimerasa , Polimorfismo de Nucleótido Simple , Radiografía Panorámica , Análisis de Secuencia de ADNRESUMEN
OBJECTIVE: Hereditary defects in tooth enamel formation, amelogenesis imperfecta (AI), can be non-syndromic or syndromic phenotype. Integrins are signaling proteins that mediate cell-cell and cell-extracellular matrix communication, and their involvement in tooth development is well known. The purposes of this study were to identify genetic cause of an AI family and molecular pathogenesis underlying defective enamel formation. MATERIALS AND METHODS: We recruited a Turkish family with isolated AI and performed mutational analyses to clarify the underlying molecular genetic etiology. RESULTS: Autozygosity mapping and exome sequencing identified a novel homozygous ITGB6 transversion mutation in exon 4 (c.517G>C, p.Gly173Arg). The glycine at this position in the middle of the ßI-domain is conserved among a wide range of vertebrate orthologs and human paralogs. Clinically, the enamel was generally thin and pitted with pigmentation. Thicker enamel was noted at the cervical area of the molars. CONCLUSIONS: In this study, we identified a novel homozygous ITGB6 mutation causing isolated AI, and this advances the understanding of normal and pathologic enamel development.
Asunto(s)
Amelogénesis Imperfecta/etiología , Amelogénesis Imperfecta/genética , Cadenas beta de Integrinas/genética , Mutación , Amelogénesis/genética , Amelogénesis Imperfecta/sangre , Amelogénesis Imperfecta/diagnóstico por imagen , Niño , Análisis Mutacional de ADN , Exones , Femenino , Humanos , Diente Molar/patología , Linaje , Reacción en Cadena de la Polimerasa , Alineación de Secuencia , TurquíaRESUMEN
Amelogenesis imperfecta (AI) is a genetic disease affecting tooth enamel formation. AI can be an isolated entity or a phenotype of syndromes. To date, more than 10 genes have been associated with various forms of AI. We have identified 2 unrelated Turkish families with hypoplastic AI and performed mutational analysis. Whole-exome sequencing identified 2 novel heterozygous nonsense mutations in the ENAM gene (c.454G>T p.Glu152* in family 1, c.358C>T p.Gln120* in family 2) in the probands. Affected individuals were heterozygous for the mutation in each family. Segregation analysis within each family revealed individuals with incomplete penetrance or extremely mild enamel phenotype, in spite of having the same mutation with the other affected individuals. We believe that these findings will broaden our understanding of the clinical phenotype of AI caused by ENAM mutations.
Asunto(s)
Amelogénesis Imperfecta/genética , Codón sin Sentido/genética , Proteínas de la Matriz Extracelular/genética , Penetrancia , Adolescente , Niño , Segregación Cromosómica/genética , Codón de Terminación/genética , Consanguinidad , Citosina , Esmalte Dental/patología , Hipoplasia del Esmalte Dental/genética , Exoma/genética , Exones/genética , Femenino , Ácido Glutámico/genética , Glutamina/genética , Guanina , Heterocigoto , Humanos , Masculino , Fenotipo , TiminaRESUMEN
Dental enamel formation depends upon the transcellular transport of Ca(2+) by ameloblasts, but little is known about the molecular mechanism, or even if the same process is operative during the secretory and maturation stages of amelogenesis. Identifying mutations in genes involved in Ca(2+) homeostasis that cause inherited enamel defects can provide insights into the molecular participants and potential mechanisms of Ca(2+) handling by ameloblasts. Stromal Interaction Molecule 1 (STIM1) is an ER transmembrane protein that activates membrane-specific Ca(2+) influx in response to the depletion of ER Ca(2+) stores. Solute carrier family 24, member 4 (SLC24A4), is a Na(+)/K(+)/Ca(2+) transporter that exchanges intracellular Ca(2+) and K(+) for extracellular Na(+). We identified a proband with syndromic hypomaturation enamel defects caused by a homozygous C to T transition (g.232598C>T c.1276C>T p.Arg426Cys) in STIM1, and a proband with isolated hypomaturation enamel defects caused by a homozygous C to T transition (g.124552C>T; c.437C>T; p.Ala146Val) in SLC24A4. Immunohistochemistry of developing mouse molars and incisors showed positive STIM1 and SLC24A4 signal specifically in maturation-stage ameloblasts. We conclude that enamel maturation is dependent upon STIM1 and SLC24A4 function, and that there are important differences in the Ca(2+) transcellular transport systems used by secretory- and maturation-stage ameloblasts.
Asunto(s)
Amelogénesis/fisiología , Antiportadores/fisiología , Proteínas de la Membrana/fisiología , Proteínas de Neoplasias/fisiología , Alanina/genética , Ameloblastos/fisiología , Amelogénesis/genética , Animales , Antiportadores/genética , Arginina/genética , Señalización del Calcio/fisiología , Niño , Preescolar , Consanguinidad , Cisteína/genética , Citosina , Hipoplasia del Esmalte Dental/genética , Femenino , Variación Genética/genética , Homocigoto , Humanos , Proteínas de la Membrana/genética , Ratones , Mutación Missense/genética , Proteínas de Neoplasias/genética , Linaje , Molécula de Interacción Estromal 1 , Timina , Valina/genéticaRESUMEN
Amelogenesis imperfecta is a heterogeneous group of genetic conditions affecting enamel formation. Recently, mutations in solute carrier family 24 member 4 (SLC24A4) have been identified to cause autosomal recessive hypomaturation amelogenesis imperfecta. We recruited a consanguineous family with hypomaturation amelogenesis imperfecta with generalized brown discoloration. Sequencing of the candidate genes identified a 10-kb deletion, including exons 15, 16, and most of the last exon of the SLC24A4 gene. Interestingly, this deletion was caused by homologous recombination between two 354-bp-long homologous sequences located in intron 14 and the 3' UTR. This is the first report of exonal deletion in SLC24A4 providing confirmatory evidence that the function of SLC24A4 in calcium transport has a crucial role in the maturation stage of amelogenesis.
Asunto(s)
Amelogénesis Imperfecta/genética , Antiportadores/genética , Exones/genética , Eliminación de Gen , Regiones no Traducidas 3'/genética , Emparejamiento Base/genética , Niño , Hibridación Genómica Comparativa , Consanguinidad , Femenino , Homocigoto , Humanos , Intrones/genética , LinajeRESUMEN
AIM: Evidence suggests caries experience is higher in children with asthma. This study compared caries experience in asthmatic and non-asthmatic children and defined whether variation in the distribution of caries experience differed between the two groups and was dependent on the presence of genetic variation in enamel formation genes. METHODS: Children with asthma were recruited at the Istanbul University, Faculty of Medicine, Department of Paediatrics, Division of Paediatric Allergy and Pulmonary Diseases, and non-affected children were recruited at the Istanbul University, Faculty of Dentistry, Department of Paedodontics. Cases (N = 100) were defined as children between the ages of 6 and 12 years with asthma and controls (N = 100) as children without asthma. Cases and controls were matched by sex and age. All study subjects received a complete dental exam, provided demographic and other caries and asthma risk factors data, and a saliva sample for DNA extraction. Caries experience was defined based on DMFT/dmft and DMFS/dmfs scores. Genotypes of 11 SNPs were selected in intronic regions of enamel development genes. PCR with TaqMan chemistry was used for genotyping all selected markers. Association between caries experience (caries-free versus caries affected) depending on asthma status and SNPs was tested with PLINK by logistic regression, adjusting by risk, and other preventive measures. p values below 0.0045 (0.05/11) were considered statistically significant. RESULTS: Logistic regression analysis showed an association between AMBN rs4694075 and caries experience (p = 2.525e-007). CONCLUSIONS: This study provides, for the first time, evidence that ameloblastin is associated with caries in asthmatic children.
Asunto(s)
Asma/complicaciones , Índice CPO , Proteínas del Esmalte Dental/genética , Variación Genética/genética , Amelogénesis/genética , Amelogenina/genética , Estudios de Casos y Controles , Niño , Caries Dental/complicaciones , Caries Dental/genética , Índice de Placa Dental , Proteínas de la Matriz Extracelular/genética , Femenino , Genotipo , Humanos , Intrones/genética , Masculino , Proteínas Nucleares/genética , Polimorfismo de Nucleótido Simple/genética , Factores de Empalme de ARN , Timina , Cepillado DentalRESUMEN
The runt-related transcription factor 2 gene (RUNX2), which is also known as CBFA1, is a master regulatory gene in bone formation. Mutations in RUNX2 have been identified in cleidocranial dysplasia (CCD) patients. CCD is a rare autosomal dominant skeletal dysplasia that is characterized by delayed closure of cranial sutures, aplastic or hypoplastic clavicle formation, short stature, and dental anomalies, including malocclusion, supernumerary teeth, and delayed eruption of permanent teeth. In this study, we recruited three de novo CCD families and performed mutational analysis of the RUNX2 gene as a candidate gene approach. The mutational study revealed three disease-causing mutations: a missense mutation (c.674G>A, p.Arg225Gln), a frameshift mutation (c.1119delC, p.Arg374Glyfs*), and a nonsense mutation (c.1171C>T, p.Arg391*). Clinical examination revealed a unique dental phenotype (no typical supernumerary teeth, but duplication of anterior teeth) in one patient. We believe that this finding will broaden the understanding of the mechanism of supernumerary teeth formation and CCD-related phenotypes.
Asunto(s)
Displasia Cleidocraneal/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Adolescente , Secuencia de Bases , Niño , Displasia Cleidocraneal/diagnóstico por imagen , Codón sin Sentido , Análisis Mutacional de ADN , Femenino , Mutación del Sistema de Lectura , Estudios de Asociación Genética , Humanos , Maxilares/diagnóstico por imagen , Masculino , Mutación Missense , Radiografía , Anomalías Dentarias/diagnóstico por imagen , Anomalías Dentarias/genéticaRESUMEN
Amelogenesis imperfecta (AI) can be either isolated or part of a larger syndrome. Junctional epidermolysis bullosa (JEB) is a collection of autosomal-recessive disorders featuring AI associated with skin fragility and other symptoms. JEB is a recessive syndrome usually caused by mutations in both alleles of COL17A1, LAMA3, LAMB3, or LAMC2. In rare cases, heterozygous carriers in JEB kindreds display enamel malformations in the absence of skin fragility (isolated AI). We recruited two kindreds with autosomal-dominant amelogenesis imperfecta (ADAI) characterized by generalized severe enamel hypoplasia with deep linear grooves and pits. Whole-exome sequencing of both probands identified novel heterozygous mutations in the last exon of LAMB3 that likely truncated the protein. The mutations perfectly segregated with the enamel defects in both families. In Family 1, an 8-bp deletion (c.3446_3453del GACTGGAG) shifted the reading frame (p.Gly 1149Glufs*8). In Family 2, a single nucleotide substitution (c.C3431A) generated an in-frame translation termination codon (p.Ser1144*). We conclude that enamel formation is particularly sensitive to defects in hemidesmosome/basement-membrane complexes and that syndromic and non-syndromic forms of AI can be etiologically related.
Asunto(s)
Amelogénesis Imperfecta/genética , Moléculas de Adhesión Celular/genética , Niño , Codón sin Sentido , Análisis Mutacional de ADN , Femenino , Mutación del Sistema de Lectura , Genes Dominantes , Humanos , KalininaRESUMEN
AIM: This study was designed to determine the type and amount of the monomers leached from the different particle sizes of the composite materials. MATERIALS AND METHODS: Three different disk sizes (2, 4, 6 mm) prepared for each material group (Filtek Flow, Filtek A110, Filtek P60 and Filtek Supreme) were polymerised by LED and halogen light; the specimens were then placed in artificial saliva. The monomer release in 30 min and 24 hrs from the specimens was analyzed in HPLC calibrated for the monomer extracts before. RESULTS: TEGDMA release was detected in all material groups after 30 min and after 24 hrs. BisGMA and BisEMA were not determined in any groups and UDMA was detected only in Filtek Supreme. Significant differences in release of TEGDMA and UDMA were obtained between the different sizes of discs. Significantly high amount of TEGDMA and UDMA monomer release was obtained in LED than Halogen groups. Lower amount of monomer release was obtained in species of 30 min than 24 hrs. CONCLUSION: Data has revealed that the monomer release could be detected significantly high from the composite materials polymerized by a lower output curing light device; and higher elution of monomers was determined as the composite thickness has increased. Therefore, the clinical applications of composite materials and the type of curing units have very important effects on the success of restorations and in the decrease of potential side effects.
Asunto(s)
Resinas Compuestas/química , Curación por Luz de Adhesivos Dentales , Metacrilatos/análisis , Bisfenol A Glicidil Metacrilato/análisis , Cromatografía Líquida de Alta Presión , Luces de Curación Dental , Tamaño de la Partícula , Polietilenglicoles/análisis , Polimerizacion , Ácidos Polimetacrílicos/análisis , Poliuretanos/análisis , Saliva Artificial , SemiconductoresRESUMEN
The proven candidate genes for amelogenesis imperfecta (AI) are AMELX, ENAM, MMP20, KLK4, FAM83H, and WDR72. We performed mutation analyses on seven families with hypomaturation AI. A novel WDR72 dinucleotide deletion mutation (g.57,426_57,427delAT; c.1467_ 1468delAT; p.V491fsX497) was identified in both alleles of probands from Mexico and Turkey. Haplotype analyses showed that the mutations arose independently in the two families. The disease perfectly segregated with the genotype. Only persons with both copies of the mutant allele were affected. Their hypomineralized enamel suffered attrition and orange-brown staining following eruption. Expression of WDR72 fused to green fluorescent protein showed a cytoplasmic localization exclusively and was absent from the nucleus. We conclude that WDR72 is a cytoplasmic protein that is critical for dental enamel formation.
Asunto(s)
Amelogénesis Imperfecta/genética , Citoplasma/ultraestructura , Proteínas/genética , Eliminación de Secuencia/genética , Adenina , Alelos , Codón sin Sentido/genética , Esmalte Dental/patología , Exones/genética , Genotipo , Proteínas Fluorescentes Verdes , Haplotipos/genética , Homocigoto , Humanos , Intrones/genética , Microscopía Confocal , Microscopía Fluorescente , Linaje , Polimorfismo de Nucleótido Simple/genética , Análisis de Secuencia de ADN , Timina , Atrición Dental/genética , Decoloración de Dientes/genética , Valina/genéticaRESUMEN
Proteolytic enzymes serve important functions during dental enamel formation, and mutations in the kallikrein 4 (KLK4) and enamelysin (MMP20) genes cause autosomal-recessive amelogenesis imperfecta (ARAI). So far, only 1 KLK4 and 3 MMP20 mutations have been reported in ARAI kindreds. To determine whether ARAI in a family with a hypomaturation-type enamel defect is caused by mutations in the genes encoding enamel proteolytic enzymes, we performed mutational analysis on candidate genes. Mutational and haplotype analyses revealed an ARAI-causing point mutation (c.910G>A, p.A304T) in exon 6 of MMP20 that results in a single amino acid substitution in the hemopexin domain. Western blot analysis showed decreased expression of the mutant protein, but zymogram analysis demonstrated that this mutant was a functional protein. The proband and an affected brother were homozygous for the mutation, and both unaffected parents were carriers. The enamel of newly erupted teeth had normal thickness, but was chalky white and became darker with age.
Asunto(s)
Amelogénesis Imperfecta/genética , Proteínas del Esmalte Dental/genética , Esmalte Dental/patología , Hemopexina/genética , Metaloproteinasa 20 de la Matriz/genética , Mutación Puntual/genética , Adulto , Amelogénesis Imperfecta/complicaciones , Amelogénesis Imperfecta/patología , Amelogenina/genética , Estudios de Casos y Controles , Niño , Análisis Mutacional de ADN , Femenino , Haplotipos/genética , Humanos , Calicreínas/genética , Masculino , Linaje , Valores de Referencia , Decoloración de Dientes/etiología , Decoloración de Dientes/genética , Decoloración de Dientes/patologíaRESUMEN
To date, 4 unique enamelin gene (ENAM) defects have been identified in kindreds with amelogenesis imperfecta. To improve our understanding of the roles of enamelin in normal enamel formation, and to gain information related to possible genotype/phenotype correlations, we have identified 2 ENAM mutations in kindreds with hypoplastic ADAI, 1 novel (g.4806A>C, IVS6-2A>C) and 1 previously identified (g.8344delG), and have characterized the resulting enamel phenotypes. The IVS6-2A>C mutation caused a severe enamel phenotype in the proband, exhibiting horizontal grooves of severely hypoplastic enamel. The affected mother had several shallow hypoplastic horizontal grooves in the lower anterior teeth. In the case of the g.8344delG mutation, the phenotype was generalized hypoplastic enamel with shallow horizontal grooves in the middle 1/3 of the anterior teeth. In general, mutations in the human enamelin gene cause hypoplastic enamel, often with horizontal grooves, but the severity of the enamel defects is variable, even among individuals with the same mutation.