Novel WNT10A variant in a Japanese case of nonsyndromic oligodontia

Abstract

Congenital tooth agenesis is one of the most common anomalies in humans. Many genetic factors are involved in tooth development, including MSX1, PAX9, WNT10A, and LRP6. Thus, mutations in these genes can cause congenital tooth agenesis in humans. In this study, we identified a novel nonsense WNT10A variant, NM_025216.3(WNT10A_v001):c.1090A > T, which produces a C-terminal truncated gene product, p.(Lys364*), in a sporadic form of congenital tooth agenesis. The variant was not found in the healthy parents and thus was considered to cause congenital tooth agenesis in the case.

Fig. 1. Oral phenotype, pedigree trees, and nucleotide substitution in WNT10A.

A Orthopantomograms with the patient’s missing teeth indicated by yellow circles. B Pedigree tree of the patient’s family. The proband is indicated by an arrow. Affected individuals are depicted by black symbols, and unaffected family members are shown by white symbols. C Sanger sequencing showing the c.1090 A > T nucleotide substitution in exon 4 of WNT10A. A Electropherogram of I:1 (father), I:2 (mother) and II:1 (patient). D Schematic diagram of WNT, LRP5/6, and FRIZZLED molecular complexes. WNT ligands bind to two transmembrane proteins: LRP5/6 and FRIZZLED. The cysteine-rich domain (CR-domain) of FRIZZLED is a binding domain for WNT ligands.

Fig. 2. Structural models and biological activity of wild-type WNT10A and the p.Lys364* variant.

A, a Amino acid sequence of the C-termini of human WNT3 and WNT10A. The truncated region is indicated by the line. The conserved amino acid residues are marked with a dash above the sequence and the cysteine residues are in red. b Structural model of WNT10A. The truncated region in the C-terminus anti-parallel beta-sheet of the WNT variant is indicated in silver gray and by the double-headed arrow. c The structure of WNT is indicated by green ribbon model and the CR-domain of FRIZZLED is shown as a red sphere model. The models were created with PyMOL Molecular Graphics System. The CR-domain of FRIZZLED interacts with the WNT ligand via the C-terminus beta-sheet structure. d, e The C-terminal deletion variant of WNT10A lacks an interaction arm and does not bind the CR-domain of FRIZZLED. B Luciferase assay for WNT/beta-catenin signaling. The WNT10A p.Lys364* variant has lost biological activity relative to the negative control level. C Western blotting of WNT10A and the variant from the transfectant cell lysate (upper panel); beta-actin served as a control (lower panel). The experiment was performed in triplicate. Lane 1, wild-type human WNT10A; Lane 2, human WNT10A p.Lys364* variant; Lane 3, mock transfection with pcDNA3.1. The degradation product is detected in the p.Lys364* WNT10A variant protein samples. The beta-actin control experiment confirmed the equality of expression levels in each sample.