RESUMEN
Clefts of the palate and/or lip are among the most common human craniofacial malformations and involve multiple genetic and environmental factors. Defects can only be corrected surgically and require complex life-long treatments. Our studies utilized the well-characterized Pax9-/- mouse model with a consistent cleft palate phenotype to test small-molecule Wnt agonist therapies. We show that the absence of Pax9 alters the expression of Wnt pathway genes including Dkk1 and Dkk2, proven antagonists of Wnt signaling. The functional interactions between Pax9 and Dkk1 are shown by the genetic rescue of secondary palate clefts in Pax9-/-Dkk1f/+;Wnt1Cre embryos. The controlled intravenous delivery of small-molecule Wnt agonists (Dkk inhibitors) into pregnant Pax9+/- mice restored Wnt signaling and led to the growth and fusion of palatal shelves, as marked by an increase in cell proliferation and osteogenesis in utero, while other organ defects were not corrected. This work underscores the importance of Pax9-dependent Wnt signaling in palatogenesis and suggests that this functional upstream molecular relationship can be exploited for the development of therapies for human cleft palates that arise from single-gene disorders.
Asunto(s)
Fisura del Paladar/genética , Factores de Transcripción Paired Box/genética , Hueso Paladar/embriología , Proteína Wnt1/agonistas , Proteína Wnt1/genética , Animales , Tipificación del Cuerpo , Proliferación Celular , Femenino , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Péptidos y Proteínas de Señalización Intercelular/genética , Masculino , Ratones , Ratones Transgénicos , Morfogénesis , Osteogénesis , Factor de Transcripción PAX9 , Fenotipo , Unión Proteica , Vía de Señalización WntRESUMEN
Mutations in the WNT10A gene were first detected in the rare syndrome odonto-onycho-dermal dysplasia (OODD, OMIM257980) but have now also been found to cause about 35-50% of selective tooth agenesis (STHAG4, OMIM150400), a common disorder that mostly affects the permanent dentition. In our random sample of tooth agenesis patients, 40% had at least one mutation in the WNT10A gene. The WNT10A Phe228Ile variant alone reached an allele frequency of 0.21 in the tooth agenesis cohort, about 10 times higher than the allele frequency reported in large SNP databases for Caucasian populations. Patients with bi-allelic WNT10A mutations have severe tooth agenesis while heterozygous individuals are either unaffected or have a mild phenotype. Mutations in the coding areas of the WNT10B gene, which is co-expressed with WNT10A during odontogenesis, and the WNT6 gene which is located at the same chromosomal locus as WNT10A in humans, do not contribute to the tooth agenesis phenotype.
Asunto(s)
Displasia Ectodérmica/genética , Anomalías Dentarias/genética , Proteínas Wnt/genética , Alelos , Anodoncia , Frecuencia de los Genes/genética , Heterocigoto , Humanos , Mutación/genética , Odontogénesis/genética , Fenotipo , Polimorfismo de Nucleótido Simple/genética , Diente/patologíaRESUMEN
The detection of gene mutations in patients with congenitally missing teeth is not very complicated; however, proving causality is often quite difficult. Here, we report the detection of a substitution mutation, A42P, within the prodomain of bone morphogenetic protein 4 (BMP4) in a small family with tooth agenesis and describe a functional alteration that may be responsible for the tooth phenotype. As BMP4 is essential for the development of teeth and also for many other organs, it would be of considerable interest to find a BMP4 mutation that is associated only with tooth agenesis. Our in vitro investigations revealed that the A42P mutation neither affected processing and secretion of BMP4 nor altered functional properties, such as the induction of alkaline phosphatase or signaling through Smad1/5/8 phosphorylation by the mature BMP4 ligand. However, immunofluorescence staining revealed that the prodomains of BMP4 which harbor the A42P substitution form fibrillar structures around transfected cells in culture and that this fibrillar network is significantly decreased when mutant prodomains are expressed. Our finding suggests that in vivo, BMP4 prodomain behavior might also be altered by the mutation and could influence storage or transport of mature BMP4 in the extracellular matrix of the developing tooth.
Asunto(s)
Fosfatasa Alcalina/análisis , Sustitución de Aminoácidos/genética , Anodoncia/genética , Proteína Morfogenética Ósea 4/genética , Regulación del Desarrollo de la Expresión Génica , Odontogénesis/genética , Anomalías Dentarias/genética , Animales , Proteína Morfogenética Ósea 4/metabolismo , Línea Celular , Humanos , Ratones , Mutagénesis Sitio-Dirigida , Mutación Missense , Plásmidos , Análisis de Secuencia de ADN , Anomalías Dentarias/enzimologíaRESUMEN
An epidemiologic study from the year 2008 found a highly significant increase of congenital tooth agenesis in women with ovarian cancer suggesting that a common genetic etiology may predispose women to both conditions. The finding was reminiscent of a previously described family harboring an AXIN2 mutation which could be shown to segregate with both the tooth agenesis and the predisposition to colon cancer transmitted in this family. Since tooth agenesis as a marker for susceptibility to ovarian cancer would be of great relevance to both oncologists and women with inborn missing teeth, the relationship between the two disorders requires a thorough assessment. We examined DNA samples from the ovarian cancer patients who participated in the original study, to look for a possible genetic connection between their ovarian malignancies and tooth agenesis. MSX1, PAX9, AXIN2, EDA, WNT10A, BARX and BRCA1 genes were selected for sequence analysis as they may cause tooth agenesis, are expressed in the female reproductive system, and/or are involved in tumorigenesis in general or specifically in the ovary. Our study revealed evidence that one half of the dually affected patients had an independent causation of the two conditions, thus reducing the previously estimated ovarian cancer risk for women with congenital tooth agenesis quite significantly.