A Putative Association of a Single Nucleotide Polymorphism in GPR126 with Aggressive Periodontitis in a Japanese Population.

Periodontitis is an inflammatory disease causing loss of tooth-supporting periodontal tissue. Disease susceptibility to the rapidly progressive form of periodontitis, aggressive periodontitis (AgP), appears to be influenced by genetic risk factors. To identify these in a Japanese population, we performed whole exome sequencing of 41 unrelated generalized or localized AgP patients. We found that AgP is putatively associated with single nucleotide polymorphism (SNP) rs536714306 in the G-protein coupled receptor 126 gene, GPR126 [c.3086 G>A (p.Arg1029Gln)]. Since GPR126 activates the cAMP/PKA signaling pathway, we performed cAMP ELISA analysis of cAMP concentrations, and found that rs536714306 impaired the signal transactivation of GPR126. Moreover, transfection of human periodontal ligament (HPDL) cells with wild-type or mutant GPR126 containing rs536714306 showed that wild-type GPR126 significantly increased the mRNA expression of bone sialoprotein, osteopontin, and Runx2 genes, while mutant GPR126 had no effect on the expression of these calcification-related genes. The increase in expression of these genes was through the GPR126-induced increase of bone morphogenic protein-2, inhibitor of DNA binding (ID) 2, and ID4 expression. These data indicate that GPR126 might be important in maintaining the homeostasis of periodontal ligament tissues through regulating the cytodifferentiation of HPDL cells. The GPR126 SNP rs536714306 negatively influences this homeostasis, leading to the development of AgP, suggesting that it is a candidate genetic risk factor for AgP in the Japanese population

List of 10 filtered variants.

<p>List of 10 filtered variants.</p

GWAS findings.

<p>GWAS findings.</p

Clinical characteristic of individual subjects in the GWAS.

<p>Clinical characteristic of individual subjects in the GWAS.</p

Effects of the SNP rs536714306 on <i>GPR126</i> signal transduction in HEK293T cells.

<p>(A) HEK293T cells were transfected with empty vector, wild-type GPR126, or mutant GPR126 containing SNP rs536714306 for 24 h, and expression of GPR126 and β-Actin was assessed by western blotting. Asterisk represents a non-specific band. (B) HEK293T cells were transfected with empty vector, wild-type GPR126, or mutant GPR126 for 48 h. Following a 1 h incubation with 3 μg/mL type IV collagen, cAMP expression was assessed by ELISA. Data represent the average and standard deviation of three independent experiments. *<i>p</i> <0.05. EV, empty vector; WT, wild-type GPR126; mut, mutant GPR126.</p

Primer list for real-time PCR.

<p>Primer list for real-time PCR.</p

Expression of GPR126 during the cytodifferentiation of HPDL cells.

<p>HPDL cells were transfected with empty vector and incubated with mineralization medium for 4, 6, 9, and 15 days and the expression of BSP and GPR126 was assessed by real-time PCR. Data represent the average and standard deviation of four independent experiments. *<i>p</i> <0.05.</p