RESUMEN
Background: The role of microRNAs (miRNAs) in animal models of palatogenesis has been shown, but only limited research has been carried out in humans. To date, no miRNA expression study on tissues or cells from cleft palate patients has been published. We compared miRNA expression in palatal fibroblasts from cleft palate patients and age-matched controls. Material and Methods: Cultured palatal fibroblasts from 10 non-syndromic cleft lip and palate patients (nsCLP; mean age: 18 ± 2 months), 5 non-syndromic cleft palate only patients (nsCPO; mean age: 17 ± 2 months), and 10 controls (mean age: 24 ± 5 months) were analysed with next-generation small RNA sequencing. All subjects are from Western European descent. Sequence reads were bioinformatically processed and the differentially expressed miRNAs were technically validated using quantitative reverse-transcription polymerase chain reaction (RT-qPCR). Results: Using RNA sequencing, three miRNAs (hsa-miR-93-5p, hsa-miR-18a-5p, and hsa-miR-92a-3p) were up-regulated and six (hsa-miR-29c-5p, hsa-miR-549a, hsa-miR-3182, hsa-miR-181a-5p, hsa-miR-451a, and hsa-miR-92b-5p) were down-regulated in nsCPO fibroblasts. One miRNA (hsa-miR-505-3p) was down-regulated in nsCLP fibroblasts. Of these, hsa-miR-505-3p, hsa-miR-92a, hsa-miR-181a, and hsa-miR-451a were also differentially expressed using RT-PCR with a higher fold change than in RNAseq. Limitations: The small sample size may limit the value of the data. In addition, interpretation of the data is complicated by the fact that biopsy samples are taken after birth, while the origin of the cleft lies in the embryonic period. This, together with possible effects of the culture medium, implies that only cell-autonomous genetic and epigenetic differences might be detected. Conclusions: For the first time, we have shown that several miRNAs appear to be dysregulated in palatal fibroblasts from patients with nsCLP and nsCPO. Furthermore, large-scale genomic and expression studies are needed to validate these findings.
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Fisura del Paladar/genética , Fibroblastos/metabolismo , MicroARNs/genética , Paladar Duro/metabolismo , Estudios de Casos y Controles , Células Cultivadas , Preescolar , Fisura del Paladar/patología , Femenino , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica , Humanos , Lactante , Masculino , Paladar Duro/patología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodosRESUMEN
Retinoic acid (RA), the active derivative of vitamin A, is one of the major regulators of embryonic development, including the development of the epidermis, the limbs and the secondary palate. In the embryo, RA levels are tightly regulated by the activity of RA synthesizing and degrading enzymes. Aberrant RA levels due to genetic variations in RA metabolism pathways contribute to congenital malformations in these structures. In vitro and in vivo studies provide considerable evidence on the effects of RA and its possible role in the development of the epidermis, the limbs and the secondary palate. In conjunction with other regulatory factors, RA seems to stimulate the development of the epidermis by inducing proliferation and differentiation of ectodermal cells into epidermal cells. In the limbs, the exact timing of RA location and level is crucial to initiate limb bud formation and to allow chondrogenesis and subsequent osteogenesis. In the secondary palate, the correct RA concentration is a key factor for mesenchymal cell proliferation during palatal shelf outgrowth, elevation and adhesion, and finally to allow bone formation in the hard palate. These findings are highly relevant to understanding the mechanism of RA signalling in development and in the aetiology of specific congenital diseases.
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Diferenciación Celular/efectos de los fármacos , Desarrollo Embrionario/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Tretinoina/administración & dosificación , Animales , Diferenciación Celular/genética , Proliferación Celular/efectos de los fármacos , Desarrollo Embrionario/genética , Extremidades/crecimiento & desarrollo , Ratones , Hueso Paladar/crecimiento & desarrolloRESUMEN
PURPOSE: We aimed to identify a novel genetic cause of tooth agenesis (TA) and/or orofacial clefting (OFC) by combining whole-exome sequencing (WES) and targeted resequencing in a large cohort of TA and OFC patients. METHODS: WES was performed in two unrelated patients: one with severe TA and OFC and another with severe TA only. After deleterious mutations were identified in a gene encoding low-density lipoprotein receptor-related protein 6 (LRP6), all its exons were resequenced with molecular inversion probes in 67 patients with TA, 1,072 patients with OFC, and 706 controls. RESULTS: We identified a frameshift (c.4594delG, p.Cys1532fs) and a canonical splice-site mutation (c.3398-2A>C, p.?) in LRP6, respectively, in the patient with TA and OFC and in the patient with severe TA only. The targeted resequencing showed significant enrichment of unique LRP6 variants in TA patients but not in nonsyndromic OFC patients. Of the five variants in patients with TA, two affected the canonical splice site and three were missense variants; all variants segregated with the dominant phenotype, and in one case the missense mutation occurred de novo. CONCLUSION: Mutations in LRP6 cause TA in humans.Genet Med 18 11, 1158-1162.
Asunto(s)
Anodoncia/genética , Exoma/genética , Predisposición Genética a la Enfermedad , Proteína-6 Relacionada a Receptor de Lipoproteína de Baja Densidad/genética , Adolescente , Anodoncia/patología , Niño , Femenino , Mutación del Sistema de Lectura/genética , Humanos , Masculino , Mutación Missense/genética , Linaje , Análisis de Secuencia de ADN , Vía de Señalización Wnt/genéticaRESUMEN
Individuals with cleft lip and palate present significantly more dental anomalies, even outside the cleft area, than do individuals without clefts. Our aim was to evaluate the prevalence of tooth agenesis and patterns of hypodontia in a large sample of patients with complete bilateral cleft lip and palate (BCLP). Serial panoramic radiographs (the first radiograph was taken at 10.5-13.5 yr of age) of 240 patients with BCLP (172 male patients, 68 female patients) were examined. Third molars were not included in the evaluation. Agenesis of at least one tooth was present in 59.8% of patients. Upper laterals and upper and lower second premolars were missing most frequently. Using the tooth agenesis code (TAC), 52 different agenesis patterns were identified, of which simultaneous agenesis of 12, 22, 15, 25, 35, and 45 was the most frequent pattern. Nine of the 240 patients showed combined BCLP and oligodontia.
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Anodoncia/etiología , Labio Leporino/complicaciones , Fisura del Paladar/complicaciones , Adolescente , Distribución de Chi-Cuadrado , Niño , Femenino , Humanos , Masculino , Variaciones Dependientes del Observador , Estudios RetrospectivosRESUMEN
The Msx1 transcription factor is involved in multiple epithelial-mesenchymal interactions during vertebrate embryogenesis. It has pleiotropic effects in several tissues. In humans, MSX1 variants have been related to tooth agenesis, orofacial clefting, and nail dysplasia. We correlate all MSX1 disease causing variants to phenotypic features to shed light on this hitherto unclear association. MSX1 truncations cause more severe phenotypes than in-frame variants. Mutations in the homeodomain always cause tooth agenesis with or without other phenotypes while mutations outside the homeodomain are mostly associated with non-syndromic orofacial clefts. Downstream effects can be further explored by the edgetic perturbation model. This information provides new insights for genetic diagnosis and for further functional analysis of MSX1 variants.
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Anodoncia/genética , Factor de Transcripción MSX1/genética , Anomalías de la Boca/genética , Mutación , Animales , Anodoncia/diagnóstico , Estudios de Asociación Genética , Humanos , Factor de Transcripción MSX1/metabolismo , Anomalías de la Boca/diagnóstico , Uñas Malformadas/diagnóstico , Uñas Malformadas/genética , SíndromeRESUMEN
Loss-of-function variants in ANKRD11 were identified as the cause of KBG syndrome, an autosomal dominant syndrome with specific dental, neurobehavioural, craniofacial and skeletal anomalies. We present the largest cohort of KBG syndrome cases confirmed by ANKRD11 variants reported so far, consisting of 20 patients from 13 families. Sixteen patients were molecularly diagnosed by Sanger sequencing of ANKRD11, one familial case and three sporadic patients were diagnosed through whole-exome sequencing and one patient was identified through genomewide array analysis. All patients were evaluated by a clinical geneticist. Detailed orofacial phenotyping, including orthodontic evaluation, intra-oral photographs and orthopantomograms, was performed in 10 patients and revealed besides the hallmark feature of macrodontia of central upper incisors, several additional dental anomalies as oligodontia, talon cusps and macrodontia of other teeth. Three-dimensional (3D) stereophotogrammetry was performed in 14 patients and 3D analysis of patients compared with controls showed consistent facial dysmorphisms comprising a bulbous nasal tip, upturned nose with a broad base and a round or triangular face. Many patients exhibited neurobehavioural problems, such as autism spectrum disorder or hyperactivity. One-third of patients presented with (conductive) hearing loss. Congenital heart defects, velopharyngeal insufficiency and hip anomalies were less frequent. On the basis of our observations, we recommend cardiac assessment in children and regular hearing tests in all individuals with a molecular diagnosis of KBG syndrome. As ANKRD11 is a relatively common gene in which sequence variants have been identified in individuals with neurodevelopmental disorders, it seems an important contributor to the aetiology of both sporadic and familial cases.
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Anomalías Múltiples/genética , Trastorno del Espectro Autista/genética , Enfermedades del Desarrollo Óseo/genética , Cromosomas Humanos Par 16 , Eliminación de Gen , Discapacidad Intelectual/genética , Proteínas Represoras/genética , Anomalías Dentarias/genética , Anomalías Múltiples/diagnóstico , Anomalías Múltiples/patología , Adolescente , Adulto , Trastorno del Espectro Autista/complicaciones , Trastorno del Espectro Autista/diagnóstico , Trastorno del Espectro Autista/patología , Enfermedades del Desarrollo Óseo/complicaciones , Enfermedades del Desarrollo Óseo/diagnóstico , Enfermedades del Desarrollo Óseo/patología , Niño , Preescolar , Análisis Mutacional de ADN , Exoma , Facies , Femenino , Expresión Génica , Genotipo , Humanos , Discapacidad Intelectual/complicaciones , Discapacidad Intelectual/diagnóstico , Discapacidad Intelectual/patología , Masculino , Persona de Mediana Edad , Fenotipo , Anomalías Dentarias/complicaciones , Anomalías Dentarias/diagnóstico , Anomalías Dentarias/patologíaRESUMEN
BACKGROUND: Current guidelines for evaluating cleft palate treatments are mostly based on two-dimensional (2D) evaluation, but three-dimensional (3D) imaging methods to assess treatment outcome are steadily rising. OBJECTIVE: To identify 3D imaging methods for quantitative assessment of soft tissue and skeletal morphology in patients with cleft lip and palate. DATA SOURCES: Literature was searched using PubMed (1948-2012), EMBASE (1980-2012), Scopus (2004-2012), Web of Science (1945-2012), and the Cochrane Library. The last search was performed September 30, 2012. Reference lists were hand searched for potentially eligible studies. There was no language restriction. STUDY SELECTION: We included publications using 3D imaging techniques to assess facial soft tissue or skeletal morphology in patients older than 5 years with a cleft lip with/or without cleft palate. We reviewed studies involving the facial region when at least 10 subjects in the sample size had at least one cleft type. Only primary publications were included. DATA EXTRACTION: Independent extraction of data and quality assessments were performed by two observers. RESULTS: Five hundred full text publications were retrieved, 144 met the inclusion criteria, with 63 high quality studies. There were differences in study designs, topics studied, patient characteristics, and success measurements; therefore, only a systematic review could be conducted. Main 3D-techniques that are used in cleft lip and palate patients are CT, CBCT, MRI, stereophotogrammetry, and laser surface scanning. These techniques are mainly used for soft tissue analysis, evaluation of bone grafting, and changes in the craniofacial skeleton. Digital dental casts are used to evaluate treatment and changes over time. CONCLUSION: Available evidence implies that 3D imaging methods can be used for documentation of CLP patients. No data are available yet showing that 3D methods are more informative than conventional 2D methods. Further research is warranted to elucidate it.
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Labio Leporino/patología , Fisura del Paladar/patología , Cara/patología , Huesos Faciales/patología , Imagenología Tridimensional/métodos , Sistemas de Registros Médicos Computarizados , Técnica de Colado Dental , Humanos , PubMedRESUMEN
This article describes the inter- and intra-familial phenotypic variability in four families with WNT10A mutations. Clinical characteristics of the patients range from mild to severe isolated tooth agenesis, over mild symptoms of ectodermal dysplasia, to more severe syndromic forms like odonto-onycho-dermal dysplasia (OODD) and Schöpf-Schulz-Passarge syndrome (SSPS). Recurrent WNT10A mutations were identified in all affected family members and the associated symptoms are presented with emphasis on the dentofacial phenotypes obtained with inter alia three-dimensional facial stereophotogrammetry. A comprehensive overview of the literature regarding WNT10A mutations, associated conditions and developmental defects is presented. We conclude that OODD and SSPS should be considered as variable expressions of the same WNT10A genotype. In all affected individuals, a dished-in facial appearance was observed which might be helpful in the clinical setting as a clue to the underlying genetic etiology.
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Deformidades Dentofaciales/genética , Mutación , Linaje , Proteínas Wnt/genética , Adulto , Anodoncia/diagnóstico , Anodoncia/genética , Niño , Preescolar , Deformidades Dentofaciales/diagnóstico , Glándulas Ecrinas/anomalías , Displasia Ectodérmica/diagnóstico , Displasia Ectodérmica/genética , Neoplasias de los Párpados/diagnóstico , Neoplasias de los Párpados/genética , Femenino , Variación Genética , Genotipo , Humanos , Hipotricosis/diagnóstico , Hipotricosis/genética , Queratodermia Palmoplantar/diagnóstico , Queratodermia Palmoplantar/genética , Masculino , FenotipoRESUMEN
OBJECTIVE: To characterize tooth agenesis patterns and their overall prevalence in patients with complete unilateral cleft lip and palate (CUCLP). DESIGN: Panoramic radiographs of 115 non-syndromic patients (78 males and 37 females) with CUCLP (85 patients had a cleft on the left and 30 on the right) from the Cleft Palate Craniofacial Unit in Nijmegen (The Netherlands) were evaluated. Third molars were not included in the evaluation. The Tooth Agenesis Code (TAC) was used to identify tooth agenesis patterns. RESULTS: Agenesis of at least one tooth was found in 48.7%, and agenesis outside the cleft was observed in 20.9% of patients. The lateral incisor of the maxillary cleft quadrant was the tooth most frequently missing (39.1%), followed by the maxillary lateral incisor (8.7%), and the mandibular second premolar (7.8%) in the non-cleft quadrants. Thirteen different tooth agenesis patterns were identified. Maxillary and/or maxillary and mandibular second and/or first premolars were involved in all patterns. CONCLUSION: A higher prevalence of tooth agenesis is observed in patients with CUCLP, even outside the cleft region, compared with the general population. Thirteen different patterns were observed, of which 6 were unique patterns. Certain teeth were involved in all agenesis patterns. Both the prevalence of orofacial clefting as well as hypodontia is more frequently observed on the left side.
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Anodoncia/epidemiología , Labio Leporino/epidemiología , Fisura del Paladar/epidemiología , Anodoncia/clasificación , Diente Premolar/anomalías , Estudios de Cohortes , Femenino , Humanos , Incisivo/anomalías , Masculino , Mandíbula/patología , Maxilar/patología , Países Bajos/epidemiología , Prevalencia , Radiografía PanorámicaRESUMEN
Nutritional factors such as vitamin intake contribute to the etiology of cleft palate. Vitamin A is a regulator of embryonic development. Excess vitamin A can cause congenital malformations such as spina bifida and cleft palate. Therefore, preventive nutritional strategies are required. This review identifies putative biological mechanisms underlying the association between maternal vitamin A intake and cleft palate. Excessive vitamin A may disturb all three stages of palatogenesis: 1) during shelf outgrowth, it may decrease cell proliferation and thus prevent tissue development; 2) it may prevent shelf elevation by affecting extracellular matrix composition and hydration; and 3) during shelf fusion, it may affect epithelial differentiation and apoptosis, which precludes the formation of a continuous palate. In general, high doses of vitamin A affect palatogenesis through interference with cell proliferation and growth factors such as transforming growth factor ß and platelet-derived growth factor. The effects of lower doses of vitamin A need to be investigated in greater depth in order to improve public health recommendations.