Phenotype and molecular characterizations of a family with dentinogenesis imperfecta shields type II with a novel DSPP mutation

BACKGROUND: Dentinogenesis imperfecta (DGI), Shields type-II is an autosomal dominant genetic disease which severely affects the function of the patients’ teeth. The dentin sialophosphoprotein (DSPP) gene is considered to be the pathogenic gene of DGI-II. In this study, a DGI-II family with a novel DSPP mutation were collected, functional characteristics of DGI cells and clinical features were analyzed to better understand the genotype-phenotype relationship of this disease. METHODS: Clinical data were collected, whole exome sequencing (WES) was conducted, and Sanger sequencing was used to verify the mutation sites. Physical characteristics of the patient’s teeth were examined using scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The localization of green fluorescent protein (GFP)-fused wild-type (WT) dentin sialoprotein (DSP) and its variant were evaluated via an immunocytochemistry (ICC) assay. The behaviors of human dental pulp stem cells (hDPSCs) were investigated by flow cytometry, osteogenic differentiation, and quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: A novel heterozygous mutation c.53T > G (p. Val18Gly) in DSPP was found in this family. The SEM results showed that the participants’ teeth had reduced and irregular dentinal tubes. The EDS results showed that the Ca/P ratio of the patients’ teeth was significantly higher than that of the control group. The ICC assay showed that the mutant DSP was entrapped in the endoplasmic reticulum (ER), while the WT DSP located mainly in the Golgi apparatus. In comparison with normal cells, the patient’s cells exhibited significantly decreased mineralization ability and lower expression levels of DSPP and RUNX2. CONCLUSIONS: The c.53T > G (p. Val18Gly) DSPP variant was shown to present with rare hypoplastic enamel defects. Functional analysis revealed that this novel variant disturbs dentinal characteristics and pulp cell behavior.