Objective: The
RYK,
EPHB2, and
EPHB3 genes are attractive candidates contributing to CL/P and CPO pathogenesis. That is because both
RYK deficient mice and
Ephb2/Ephb3 (genes for interaction molecules with
RYK) double-mutant mice show cleft palate.
Setting: Mutation search on
RYK,
EPHB2 and
EPHB3 was carried out in a large number of Japanese and Vietnamese patients with CL/P and CPO. We also performed a case-control study and transmission disequilibrium test (TDT) using three single nucleotide polymorphisms (SNPs) within a linkage disequilibrium block in
RYK, and seven haplotypes that were constructed from the SNPs.
Results: A missense mutation, 1355G > A (Y452C), in
RYK was identified in one Vietnamese patient with CL/P. This mutation was never found among 1,646 Vietnamese/Japanese/Caucasians including 354 CL/P and CPO patients. A colony formation assay using NIH3T3 cells transfected with mutant cDNA revealed that mutant
RYK had a significantly reduced protein activity compared to those with the wild-type
RYK, implying that the transformation ability of
RYK is depleted by this mutation. Although the case-control study and TDT on three individual SNPs did not reveal any evidence for association with oral clefts, the case-control study on one rare haplotype suggested positive association in Japanese patients with CL/P and CPO. No mutations in
EPHB2 and
EPHB3 were found in any of the patients examined.
Conclusion: All of these findings suggested that a missense mutation, 1355G > A, and one rare SNP-haplotype may play a role in the development of CL/P in Vietnamese, and CL/P and CPO in the Japanese, respectively.
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