| Objective:Primary open-angle glaucoma is a complex,multi-gene genetic heterogeneous disease caused by a combination of genetic and environmental factors.Previously,GAS7,SIX1-SIX6,and TLR4 were found to be related to the pathogenicity of POAG through genome-wide association study.Among them,four SNPs including rs11656696?GAS7?,rs10483727?SIX1-SIX6?,rs4986790?TLR4?and rs49867914?TLR4?have been verified to be significantly associated with POAG in other ethnicity.Through a case-control study targeting the Chinese Han population,this study is intended to verify whether the four SNPs including rs11656696,rs10483727,rs4986790 and rs4986791 are related to the genetic susceptibility to POAG,aiming to further filter the risk alleles associated with the pathogenesis of POAG and provide genetic basis and targets for further exploration of its potential pathogenic mechanism.Methods:1.The 894 POAG patients and 997 control subjects recruited through the case-control study were all from the Chinese Han population.2.Extract the genomic DNA from the blood sample of the subject and use SNaPshot method for genotyping,which is a typing technique based on the principle of fluorescently labeled single base extension chain termination reaction.3.The obtained data were input into software SPSS 22.0 for statistical analysis.The Chi-square test and Fisher's exact test were used to verify Hardy-Weinberg equilibrium and count allele or genotype frequencies.Logistic regression model was used to analyze the allele genetic model,and the influencing factors such as gender and age between the case and the control group were adjusted.The Bonferroni method was used to correct the P values of 4 genetic models included in the study,and the statistical significance level of each SNP was set to0.0125?significant level 1/n,1/4 of 0.05?.Power and Sample Size Calculation?version 3.1.2?was used to calculate statistical power.Results:The allele frequency and genotype frequency of the SNP in the case group and the control group were consistent with HWE?P>0.05?,indicating that the study object was representative of the population.SNP genotyping call rates were all greater than 98%.Case-control statistical results showed that:1.Between cases and controls,the frequency of the A allele of rs11656696A/T?GAS7?was statistically significant?P=0.0064,OR=0.82,95%CI=0.72-0.95?.There are also significant statistical differences between the case group and the control group in homozygous model?AA vs CC,P=0.0064,OR=0.6895%CI=0.51-0.90?and recessive model?AA vs AC+CC,P=0.00034,OR=0.66,95%CI=0.52-0.84?.2.The C allele frequency of rs10483727 C/T?SIX1-SIX6?between the case group and the control group was significantly different?P=0.032,OR=0.84,95%CI=0.72-0.99?.Between cases and controls,there are significant statistical differences in heterozygote model?CT vs TT,P=0.0033,OR=0.75,95%CI=0.62-0.91?and dominant model?CC+CT vs TT,P=0.0064,OR=0.77,95%CI=0.64-0.93?.The analysis with a total of 3246 POAG patients and 2298 controls after combining sample included in this study with samples of Chinese population in previous reports showed that rs10483727 was significantly associated with POAG(P=1.35×10-7,OR=0.78,95%CI=0.71-0.86).3.The allele frequency of rs4986790 G/A?TLR4?and rs4986791 T/C?TLR4?between POAG cases and controls were not statistically significant.And there is no statistical difference by 4 genetic models between the case group and the control group.Conclusion:This study used a case-control study to investigate the susceptibility of rs11656696?GAS7?,rs10483727?SIX1-SIX6?,rs4986790?TLR4?and rs49867914?TLR4?to the occurrence of POAG in a sample of Chinese Han population from 894 POAG patients and 997 controls by using SNaPshot method for genotyping.The corresponding statistical analysis showed that:1.The rs11656696 of the GAS7 gene might be related to the occurrence of POAG in the Chinese Han population.Given that homozygous model and recessive model was statistically different between POAG cases and controls,it suggested that subjects carrying the rs11656696"AA"genotype had a lower risk of POAG than subjects carrying the"AC+CC"genotype,speculating that the A allele of rs11656696 might be involved in reducing the risk of POAG susceptibility.2.The rs10483727 in the SIX1-SIX6 region might be related to the occurrence of POAG in the Chinese Han population.Given that heterozygote model and dominant model was statistically different between POAG cases and controls,it suggested that subjects carrying the rs10483727"CT"genotype and rs10483727"CC+CT"genotype had a lower risk of POAG than subjects carrying the"TT"genotype.Further increasing the sample size,the analysis of the sample included in this study and the sample of Chinese population in previous reports showed that individuals carrying the C allele of rs10483727 had a lower risk of POAG than individuals carrying the T allele of rs10483727,speculating that the C allele of rs10483727 might be involved in reducing the risk of POAG susceptibility.3.PS results showed that the power of test of the four SNPs were all lower than80%,suggesting that this study needed to increase the sample size to improve the mining of potential and trending genotypes and disease phenotypes.This study filtered candidate genes GAS7,SIX1-SIX6,and TLR4 for the risk alleles associated with the pathogenesis of POAG,providing new targets for the genetic susceptibility research of POAG and genetic basis for further exploring its potential pathogenic mechanism. |
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