Splicing mutation in DSPP causes dentinogenesis imperfecta and amelogenesis imperfecta

Dentin sialophosphoprotein (DSPP) is an extracellular matrix protein, which is highly expressed in odontoblasts and transiently expressed in presecretory ameloblasts. DSPP mutations were related to dentinogenesis imperfecta (DGI), some of which can be accompanied by amelogenesis imperfecta (AI). However, the mechanism underlying DGI and AI caused by DSPP mutations is still unclarified. This study aimed to reveal the molecular pathogenesis in a Chinese family with both DGI and AI caused by the DSPP splicing mutation. One Chinese family with DGI and AI was recruited. Whole-exome sequencing and Sanger sequencing were performed to identify mutations in candidate genes. Minigene splicing assays were performed to analyze the mutation effects on mRNA splicing alteration. Furthermore, point mutation (named KI) and exon 3 knocked out (named KO) mouse models were generated to understand the in vivo consequences. HE staining and microCT analysis were performed to observe the morphological changes. RNA sequencing and quantitative real-time PCR were conducted to explore the pathogenic molecular mechanism. The dentitions of the proband exhibited an opalescent color with severe attrition. Additionally, pitted enamel can be observed in the crown. A splicing mutation (NM_014208.3: c.135 + 3 A > G) of DSPP can be detected in the proband and her father. Minigene splicing assay revealed that this mutation could cause partial exon 3 of DSPP (c.246-c.255) skipping. Phenotypic analysis of mutated mice revealed enlarged pulp cavities in younger mice, and narrowed pulp chamber and canals in older mutants. Enamel abnormalities were exclusively in KI mice. RNA sequencing and quantitative real-time PCR suggested that the splicing mutation of Dspp might downregulate the expression of secreted phosphoprotein 1 (Spp1) and cartilage oligomeric matrix protein (Comp) gene, implicating the ECM-receptor interaction and focal adhesion signaling pathways in the pathogenesis of tooth abnormalities. In this study, we identified a splicing mutation in DSPP, which caused both DGI and AI. This research enhances our understanding of pathologic mechanisms of DSPP splicing mutations in tooth defects through genetic and molecular lens.