Genetic Variant in lncRNA RP11-462G12.2 Confers the Risk of Non-Syndromic Orofacial Clefts

Abstract

Background: Non-syndromic Orofacial Clefts (NSOCs) are one of the most common human birth defects. Here, we test the hypothesis that, lncRNA RP11-462G12.2 rs2262251, in high linkage disequilibrium (LD) with a newly NSOC single nucleotide polymorphism (SNP) rs8049367, was a casual SNP of NSOC. Methods: SNP rs2262251 was genotyped in a cohort of 1,321 NSOC cases and 1,274 controls, and the mechanism of rs2262251 in the development of NSOC was explored with a series of biochemical and cell molecular biology assays. Findings: We found that rs2262251 was significantly associated with the risk of NSOC and cleft lip with/without palate (CL/P), and the C allele presented a protective role against the occurrence of NSOC and CL/P. The SNP rs2262251 G allele was associated with increased expression of lncRNA RP11-462G12.2 in cells and human lip tissues. Over-expression of rs2262251 [G] contributed to reduced numbers of G0/G1 phase cells, inhibited cell apoptosis, and promoted cell proliferation. Furthermore, rs2262251 [C] could interact with miR-744-5p and regulate the expression of IQSEC2, both of which were expressed in human lip tissues and mouse craniofacial development. Interpretation: We report for the first time that lncRNA RP11-462G12.2 rs2262251 might participate in a 'lncRNA-miRNA-gene' regulatory axis during the development of NSOC. Funding Statement: National Natural Science Foundation of China (81570959, 81200808, 81230022, 81400546); Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD-2014-37); Natural Science Foundation of Jiangsu Province (BL2014073, 15KJA320002); Qing Lan Project, six distinguished talent (2016-WSW-008), Jiangsu provincial medical talent (Yongchu pan); Jiangsu provincial key medical discipline (zdxka2016026). Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: Written informed consent was obtained from each subject at recruitment. This study was approved by the institutional review board of Nanjing Medical University (Approval number: NJMUERC [2008] No. 20).