Department of Oral and Maxillo Facial Sciences, Sapienza University of Rome, Via Caserta 6, 00161, Rome, Italy.
Clinical Genomics Unit, IRCCS Casa Sollievo della Sofferenza, Viale Cappuccini 1, 71013, San Giovanni Rotondo, FG, Italy.
Arch Oral Biol. 2018 Jul;91:96-102. doi: 10.1016/j.archoralbio.2018.04.011. Epub 2018 Apr 21.
The aim of this study was the clinical and molecular characterization of a family segregating a trait consisting of a phenotype specifically involving the maxillary canines, including agenesis, impaction and ectopic eruption, characterized by incomplete penetrance and variable expressivity.
Clinical standardized assessment of 14 family members and a whole-exome sequencing (WES) of three affected subjects were performed. WES data analyses (sequence alignment, variant calling, annotation and prioritization) were carried out using an in-house implemented pipeline. Variant filtering retained coding and splice-site high quality private and rare variants. Variant prioritization was performed taking into account both the disruptive impact and the biological relevance of individual variants and genes. Sanger sequencing was performed to validate the variants of interest and to carry out segregation analysis.
Prioritization of variants "by function" allowed the identification of multiple variants contributing to the trait, including two concomitant heterozygous variants in EDARADD (c.308C>T, p.Ser103Phe) and COL5A1 (c.1588G>A, p.Gly530Ser), specifically associated with a more severe phenotype (i.e. canine agenesis). Differently, heterozygous variants in genes encoding proteins with a role in the WNT pathway were shared by subjects showing a phenotype of impacted/ectopic erupted canines.
This study characterized the genetic contribution underlying a complex trait consisting of isolated canine anomalies in a medium-sized family, highlighting the role of WNT and EDA cell signaling pathways in tooth development.
本研究旨在对一个家族的表型进行临床和分子特征分析,该表型主要涉及上颌尖牙,包括先天缺失、阻生和异位萌出,其表现为不完全外显和表现度可变。
对 14 名家族成员进行了临床标准化评估,并对 3 名受影响的个体进行了全外显子组测序(WES)。使用内部实现的管道进行 WES 数据分析(序列比对、变异调用、注释和优先级排序)。变异过滤保留了编码和剪接位点的高质量私有和罕见变异。优先考虑个体变异和基因的破坏性影响和生物学相关性。进行 Sanger 测序以验证感兴趣的变异并进行分离分析。
通过“功能”对变异进行优先排序,确定了多个导致该表型的变异,包括 EDARADD(c.308C>T,p.Ser103Phe)和 COL5A1(c.1588G>A,p.Gly530Ser)中两个同时存在的杂合变异,这些变异与更严重的表型(即犬齿先天缺失)特别相关。相反,编码在 WNT 途径中具有作用的蛋白质的基因中的杂合变异被表现为阻生/异位萌出的犬齿的个体所共有。
本研究对一个由孤立性犬齿异常组成的复杂表型的遗传贡献进行了特征描述,突出了 WNT 和 EDA 细胞信号通路在牙齿发育中的作用。