Claire Elcock

Novel missense mutations and a 288-bp exonic insertion in PAX9 in families with autosomal dominant hypodontia

We describe the molecular analysis of three families with hypodontia involving primarily molar teeth and report two novel mutational mechanisms. Linkage analysis of two large families revealed that the hypodontia was linked to the PAX9 locus. These two families revealed missense mutations consisting of a glutamic acid substitution for lysine and a proline substitution for leucine within the paired domain of PAX9. A pair of identical twins affected with hypodontia in a third family demonstrated a 288‐bp insertion within exon 2 that resulted in a putative frameshift mutation and a premature stop codon. The insertion was associated with the loss of 7‐bp from exon 2. A block of 256‐bp of sequence within the insertion was completely identical to downstream sequence from the second intron of the PAX9 gene. These studies extend the spectrum of mutations in PAX9 associated with hypodontia to include heretofore undescribed categories, including missense mutations.

Heimler Syndrome Is Caused by Hypomorphic Mutations in the Peroxisome-Biogenesis Genes PEX1 and PEX6

Heimler syndrome (HS) is a rare recessive disorder characterized by sensorineural hearing loss (SNHL), amelogenesis imperfecta, nail abnormalities, and occasional or late-onset retinal pigmentation. We ascertained eight families affected by HS and, by using a whole-exome sequencing approach, identified biallelic mutations in PEX1 or PEX6 in six of them. Loss-of-function mutations in both genes are known causes of a spectrum of autosomal-recessive peroxisome-biogenesis disorders (PBDs), including Zellweger syndrome. PBDs are characterized by leukodystrophy, hypotonia, SNHL, retinopathy, and skeletal, craniofacial, and liver abnormalities. We demonstrate that each HS-affected family has at least one hypomorphic allele that results in extremely mild peroxisomal dysfunction. Although individuals with HS share some subtle clinical features found in PBDs, the diagnosis was not suggested by routine blood and skin fibroblast analyses used to detect PBDs. In conclusion, our findings define HS as a mild PBD, expanding the pleiotropy of mutations in PEX1 and PEX6.

Tooth dimensions in hypodontia with a known PAX9 mutation

AIM Congenital absence of teeth is a complex condition affecting several parameters of oral development. This is the first study to measure tooth crown dimensions using image analysis in a family with hypodontia in whom the mutation has been identified, and compare them with a control group. METHODS AND RESULTS Study models were obtained from 10 family members from three generations affected by severe hypodontia with a missense mutation in PAX9 and 10 unaffected, unrelated controls. Using established image analysis techniques all teeth up to and including the first permanent molars were digitally imaged by two operators from the occlusal (O) and buccal (B) aspects three times and an average made for the mesio-distal (MDO and MDB) bucco-lingual (BL), area (A) and perimeter (P) measurements. Intra-class correlation coefficients (ICCC) were calculated to assess intra- and inter-operator reliability. Two-sample t-tests were then used to compare these dimensions with those of the controls. Reliability of the technique was high (mean r>0.95). The majority of tooth types throughout the dentition were significantly smaller in the family members with hypodontia than in the control group for all parameters measured. The levels of significance were very high for upper lateral incisors (p<0.0001) whilst the canines and first molars were less different. The greatest number of significant differences were found in BL and P, closely followed by MD and A measurements. CONCLUSIONS The significantly smaller tooth crown dimensions recorded in the affected family members show that the effect of the PAX9 mutation is seen not only in the congenitally missing teeth but also in smaller crown size throughout the dentition.

Incisor crown shape and crowding

The aim of this study was to investigate correlations between the shape of mandibular incisor crowns and crowding. Study models of 50 untreated white subjects (25 men and 25 women) with varying degrees of crowding were studied. The lower incisors were sectioned and imaged at the contact point and midpoint levels, and the mesiodistal width was measured. Crowding was quantified by using both Littles irregularity index and anterior tooth size-arch length discrepancy. The ratio of the 2 mesiodistal widths was correlated with Littles irregularity index and anterior tooth size-arch length discrepancy. In the women, the ratio showed correlations of 0.52 (P =.008) and 0.55 (P =.005) for central and lateral incisors, respectively, with anterior tooth size-arch length discrepancy. No predictors of lower incisor crowding could be established from mandibular incisor crown shape in this study.

The aetiology of hypodontia: The prevalence, severity and location of hypodontia within families

AIMS Previous studies have indicated that hypodontia has a significantly higher prevalence in the relatives of affected individuals than in the general population. This study aims to examine further the roles of genetic and environmental factors in the aetiology of hypodontia by investigating the relationship between the severity and distribution of hypodontia between family members, and any discernable effect of maternal health during pregnancy and birth weight. METHODS AND RESULTS 117 first degree relatives of 41 index patients were examined clinically and radiographically to identify the presence, severity and location of hypodontia. Both siblings and parents of index patients had a higher prevalence of hypodontia than the general population. The number and location of missing teeth was not related to the number and location of missing teeth in parents or siblings. The expression of hypodontia within a family was not affected by maternal health during pregnancy. CONCLUSIONS The variation found in the expression of hypodontia within families suggests that its occurrence is not solely determined by genetic factors, but epigenetic and environmental factors probably also are important. This finding is consistent with a multifactorial aetiology for this condition.

The Structure and Composition of Deciduous Enamel Affected by Local Hypoplastic Autosomal Dominant Amelogenesis Imperfecta Resulting from an ENAM Mutation

In a group of families in northern Sweden, a mutation in the ENAM gene (predicted to produce a highly truncated protein) results in the local hypoplastic form of autosomal dominant amelogenesis imperfecta. In this study, sections of deciduous teeth from members of 3 of these families were examined by scanning electron microscopy (SEM) and the enamel mineral was analysed by energy dispersive X-ray spectroscopy (EDX). The sections were also probed with antibodies raised to a conserved sequence of the enamelin protein. Selected intact teeth were first analysed by digital imaging and ascribed with an ‘Enamel Defects Index’ (EDI) score. SEM of tooth sections revealed disrupted prism morphology and the prisms had a glass-like appearance in some areas. These areas of dysplasia were sometimes irregular but formed regular arrays in others. Comparison of EDI scores with SEM indicated that in one tooth the surface had no measurable defects but significant defects were present in the underlying enamel microstructure. SEM immunohistochemistry with the antibody raised to a fragment of the enamelin protein produced positive, but light, labelling throughout normal enamel. In dysplastic areas, however, the labelling intensity appeared to be reduced. The results indicate that the presence of functional enamelin in the correct amounts is necessary for correct prism morphogenesis. In addition, a combination of EDI and structural analysis indicate that defects in enamel microstructure are not necessarily visible as defects on the surface of the tooth, suggesting the possibility, at least, that some instances of under-diagnosis may occur.

Enamel defects in extracted and exfoliated teeth from patients with Amelogenesis Imperfecta, measured using the extended enamel defects index and image analysis

Aims To enhance the phenotypic description and quantification of enamel defects from a North Sweden sample of extracted and exfoliated teeth originating from families with Amelogenesis Imperfecta by use of the extended enamel defects index (EDI) and image analysis to demonstrate the comparable reliability and value of the additional measurements. Methods and results The sample comprised 109 deciduous and 7 permanent teeth from 32 individuals of 19 families with Amelogenesis Imperfecta in Northern Sweden. A special holder for individual teeth was designed and the whole sample was examined using the extended EDI and an image analysis system. In addition to the extended EDI definitions, the calibrated images were measured for tooth surface area, defect area and percentage of surface affected using image analysis techniques. The extended EDI was assessed using weighted and unweighted Kappa statistics. The reliability of imaging and measurement was determined using Fleiss’ intra-class correlation coefficient (ICCC). Kappa values indicated good or excellent intra-operator repeatability and inter-operator reproducibility for the extended EDI. The Fleiss ICCC values indicated excellent repeatability for the image analysis measurements. Hypoplastic pits on the occlusal surfaces were the most frequent defect in this sample (82.6%). The occlusal surface displayed the most post-eruptive breakdown (39.13%) whilst the incisal portion of the buccal surfaces showed most diffuse opacities (53.4%). Image analysis methods demonstrated the largest mean hypoplastic pit areas were on the lingual surfaces. The largest mean post-eruptive breakdown areas were on the lingual surfaces of posterior teeth. The largest mean demarcated opacity areas were found on the labial surfaces. Conclusions The extended EDI and the standardised image acquisition and analysis system provided additional information to conventional measurement techniques. Additional phenotypic variables were described.

Multiple crown size variables of the upper incisors in patients with supernumerary teeth compared with controls.

AIMS As part of ongoing studies of the aetiology of dental anomalies the aims of this study were to identify multiple components of tooth size of the upper permanent incisors in 34 patients with supernumerary teeth and to compare them with those in a control group to determine whether the presence of a supernumerary tooth has a local effect on the size of the surrounding dentition. METHODS AND RESULTS The labial and occlusal aspects of the clinical crowns of the upper permanent central and lateral incisors on the study models of 74 subjects were digitally imaged and measured using an image analysis system and automated macro (34 patients with supernumerary in the upper incisor region: 17 males and 17 females and 40 controls: 20 males and 20 females). The macro-defined 17 variables from each view. From the labial view these were: the mesio-distal and occluso-gingival length and additional measurements along 25 and 75% of the mesio-distal line and at 25, 50 and 75% along the occluso-gingival line such that all these sub-divisions extended to the periphery of the tooth. From the occlusal view these were: the mesio-distal and labio-lingual lengths, and additional variables that sub-divided the mesio-distal again at 25 and 75% along the length and at 25, 50 and 75% along the labio-lingual dimension. Principal component analysis (PCA) was used to identify the key factors with the most random variability. Comparisons were then carried out between the supernumerary cases and control group using 2-way ANOVA. Seven factors of tooth size for the upper central incisors and eight factors for the upper lateral incisors were extracted. Most of these variables were found to be larger in the supernumerary group than in the control. Statistically significant differences were found for 5 out of 7 and 4 out of 8 for the upper central and lateral incisors respectively. CONCLUSIONS A number of factors of tooth size were identified and found to be larger in the supernumerary group compared to the control (7 for upper central and 8 for upper lateral incisors); the majority reached the 0.05 significance level. Tooth crown size of the upper central incisor was affected more than that of the upper lateral incisor, supporting a local field effect.