Genetic Association Study Between The COL11A1 And COL18A1 Genes And High Myopia And Function Study Of The Myopia Susceptible Gene SNTB1

Purpose:A typical eye study found that high myopia to be a characteristic feature of these mutations in collagen,type XI,alpha 1(COL11A1)and collagen,type XVIII,alpha 1(COL18A1)genes.In this study,we plan to evaluate the association between high myopia(HM)and single nucleotide polymorphisms(SNPs)in COL11A1 and COL18A1 genes in a Han Chinese population.(5 SNPs:rs17127311,rs11911327,rs2236454,rs2236457,and rs2236475).And SNTB1 gene as susceptible gene for myopia,there are few researches about the gene.This experiment plans to study myopia susceptible gene SNTB1 related function research.This study will elucidate the role of SNTB1 in myopia development and provide valuable information for the diagnosis and treatment of high myopia.Methods:A case-control association study of 869 unrelated patients with high myopia and 804 control subjects matched in ethnicity.Genomic DNA was obtained through simple-salting-out method in different NaCl concentrations.The genotyping of five SNPs in COL11A1 and COL18A1 was fulfilled by using the Snapshot method which was based on dye termination sequencing theory and ABI 3130 genetic analyzer.And Hardy-Weinberg equilibrium(HWE)was applied to test the genotypic distribution of the five SNPs.The genotyping data were analyzed using the?~2 test.And the linkage disequilibrium block structure was calculated and examined by Haploview software.All the statistical analyses were conducted by SPSS software(version 22.0).For SNTB1gene,it produces SNTB1 gene plasmid for amplification experiment.And SNTB1 gene plasmid to transfer renal epithelial cells(293T)and mouse fibroblast(3T3),by Fluorescence Microplate Reader to test cell proliferation,by Transwell to test the ability of cell migration,by Matrigel to test cell adhesion ability.Results:All of the five SNPs(in COL11A1 and COL18A1 genes)were successfully genotyped,and the genotype distributions were within HWE in both case and control groups(P>0.05).We found that rs2236475 in the COL18A1 gene was significantly associated with HM.The risk allele(G)frequency(RAF)of rs2236475was 0.215 in the controls and 0.245 in the patients,and the adjusted allelic P value was0.016,with an adjusted OR of 1.26(95%CI=1.06-1.51).However,rs2236475 showed no meaningful association with HM after the Bonferroni correction test(corrected P=0.08).The other four SNPs not showed a positive association(allelic P>0.05).Additionally,different genetic models were used to further evaluate the association between rs2236475 and HM.The frequency of the GG genotype was higher in the HM group than that in the control group(6.8%vs.4.1%,respectively).And rs2236475GG carriers had an increased risk of HM compared with rs2236475AA and rs2236475AG+AA carriers(P=0.008,OR=1.79,95%CI=1.18-2.64;P=0.012,OR=1.74,95%CI=1.12-2.57,respectively).Furthermore,haplotype analysis indicated that three SNPs(rs2236454,rs2236457 and rs2236475)of the COL18A1 gene were in the same LD block in this study,with D'from 0.84 to 0.99.The frequency of protective haplotype CTA generated from these three SNPs was significantly different between the HM cases and controls(P=0.017).With this haplotype,an individual has a 0.31-fold decrease in susceptibility to high myopia(OR=0.69,95%CI=0.51-0.94).SNTB1 gene plasmid transfect human kidney epithelial cells(293T)and mouse fibroblast(3T3),through the detection of cell proliferation,cell migration and adhesion ability.Results showed SNTB1 gene no significant influence on cell proliferation and adhesion,but it can cause the inhibition of migration(P<0.05).Conclusions:Our results suggested that common polymorphisms in COL11A1 and COL18A1 genes were unlikely to play important roles in the genetic susceptibility to high myopia.With CTA haplotype,an individual has a 0.31-fold decrease in susceptibility to high myopia(OR=0.69,95%CI=0.51-0.94).Myopia susceptible gene SNTB1 may have the inhibition of cell migration,it is no significant influence on cell proliferation and adhesion.But it is not clear of the mode of regulation and the mechanism in high myopia.Additional studies are required to verify the possible functions of these genes in the development of myopia.