Association Of CFH,LOC387715/ARMS2 And HTRA-1 Polymorphisms With Age-Related Macular Degeneration

【Purpose】To investigate association of CFH(Y402H rs1061170,-257C＞T rs3753394,IVS15 rs1329428),LOC387715/ARMS2 rs10490924,HTRA1 rs11200638 polymorphisms with exudative AMD in a northern Chinese cohort.To study the distribution of genotypes and alleles of CFH(Y402H rs 1061170,-257C＞T rs3753394,IVS15 rs1329428),LOC387715/ARMS2 rs10490924,HTRA1 rs11200638 polymorphisms in exudative AMD and healthy control group in a northern Chinese cohort and the correlation between these polymorphisms and the pathogenesis of AMD.【Method】159 patients with exudative AMD(136 for CFH,159 for LOC387715/ARMS2 and HTRA-1)and 140 healthy controls were enrolled in this study.Genomic deoxyribonucleic acid(DNA)was extracted from exudative AMD patients and healthy control subjects,then stored in -20℃.The genotypes of CFH (Y402H rs 1061170,-257C＞T rs3753394,IVS15 rs 1329428),LOC387715/ARMS2 rs10490924,HTRA1 rs11200638 were detected by polymerase chain reaction and restriction fragment length polymorphism(PCR-RFLP) methods in age-related macular degeneration patients and controls.PCR amplification products were vedficated by electrophoresis on agarose gel.Statistical analyses were performed with SPSS 13.0 software for Windows.The criterion for statistical significance was P-value＜0.05.Tests for Hardy - Weinberg equilibrium were carded out by chi-square analysis.Genotype and allele frequencies between cases and controls were compared by chi-square analysis.Haplotypes were constructed using observed genotypes by the SHEsis program,and verified by Haplov iew v4.1.【Result】1,CFH Y402H rsl061170:The frequency distribution of Y402H rs1061170 genotypes in age-related macular degeneration patients was that 2.9%of homozygous CC,55.97%of homozygous TT,and 41.2%of heterozygous CT.The distributions of C allele and T allele were 23.5%and 76.5%respectively;In healthy control group,the frequency distribution of genotypes was that 1.4%of homozygous CC,83.6%of homozygous TT,and 15.0%of heterozygous CT,and the distributions of C allele and T allele were 8.9%and 91.1%respectively.There was a significant difference between age-related macular degeneration patients and healthy control group in distribution of Y402H rs1061170 genotypes and alleles(P＜0.05).2,CFH -257C＞T rs3753394:The frequency distribution of-257C＞Trs3753394 genotypes in age-relatd macular degeneration patients was that 51.5%of homozygous TT,10.3%of homozygous CC,and 38.2%of heterozygous TC.The distributions of T allele and C allele were 70.6%and 29.4%respectively;In healthy control group,the frequency distribution of genotypes was that 33.8%of homozygous TT,17.3%of homozygous CC,and 48.9%of heterozygous TC,and the distributions of T allele and C allele were 58.3%and 41.7%respectively.There was a significant difference between age-related macular degeneration patients and healthy control group in frequencies distribution of-257C＞T rs3753394 genotypes and alleles(P＜0.05).3,CFH IVS15 rs1329428:The frequency distribution of IVS15 rs1329428 genotypes in age-related macular degeneration patients was that 39.7%of homozygous GG,14.7%of homozygous AA,and 45.6%of heterozygous GA.The distributions of G allele and A allele were 62.5%and 37.5%respectively;In healthy control group,the frequency distribution of genotypes was that 23.6%of homozygous GG,27.1%of homozygous AA,and 49.3%of heterozygous GA,and the distributions of G allele and A allele were 48.2%and 51.8%respectively.There was a significant difference between age-related macular degeneration patients and healthy control group in distribution of IVS 15 rs1329428 genotypes and alleles(P＜0.05).4,LOC387715/ARMS2 rs10490924:The frequency distribution of LOC387715/ARMS2 rs10490924 genotypes in age-related macular degeneration patients was that 10.7%of homozygous GG,43.4%of homozygous TT,and 45.9%of heterozygous GT,and the distributions of G allele and T allele were 33.6%and 66.4% respectively;In healthy control group,the frequency distribution of genotypes was that 37.9%of homozygous GG,14.3%of homozygous TT,and 47.9%of heterozygous GT,and the distributions of G allele and T allele were 61.8%and 38.2% respectively.There was a significant difference between age-related macular degeneration patients and healthy control group in distribution of LOC387715/ARMS2 rs10490924 genotypes and alleles(P＜0.05).5,HTRA-1 rs11200638:The frequency distribution of HTRA-1 rs11200638 genotypes in age-related macular degeneration patients was that 8.2%of homozygous GG,62.3%of homozygous AA,and 30%of heterozygous GA,and the distributions of G allele and A allele were 23%and 77%respectively;In healthy control group,the frequency distribution of genotypes was that 30%of homozygous GG,22.1%of homozygous AA,and 47.9%of heterozygous GA,and the distributions of G allele and A allele were 53.9%and 46.1%respectively.There was a significant difference between age-related macular degeneration patients and healthy control group in distribution ofHTRA-1 rs11200638 genotypes and alleles(P＜0.05).6,Haplotype association analysis:Haplotye TGC was observed signicantly associated with exucative AMD(OR=3.033,95%CI 1.512～6.084) in the association analysis of 3 SNPs of CFH.But,Y402H was only in low LD with-257C＞T and IVS15 (D'＜0.5,r~2＜0.3).Haplotye TA was observed signicantly associated with an increased risk of exudative AMD(OR=3.316,95%CI 2.05～5.36) in the association analysis of LOC387715/ARMS2 rs 10490924 and HTRA-1rs11200638. LOC387715/ARMS2 was in high LD with HTRA-1(D'=0.96,r~2=0.86).【Conclusions】1,CFH,LOC387715/ARMS2 and HTRA-1 gene exists a single nucleotide polymorphisms(SNPs) in the northern Chinese cohort.Our results suggest that the CFH(Y402H rs1061170,-257C＞T rs3753394,IVS15 rs1329428),LOC387715/ARMS2 rs10490924,HTRA1 rs11200638 polymorphism may be associated with age-related macular degeneration.Y402H(C/T),-257C＞T(T/C),IVS15(G/A),LOC387715/ARMS2(G/T) and HTRA-1(G/A) polymorphism are risk factors for age-related macular degeneration.2,Haplotype association analysis:Y402H was in low LD with-257C＞T and IVS15.We couldn't prove that these 3 SNPs play roles together in the pathogenesis of exudative AMD.LOC387715/ARMS2 was in high LD with HTRA-1.The haplotye TA was in significant association with an increased risk of exudative AMD (OR=3.316,95%CI2.05～5.36).3,This research suggest that both CFH(Y402H rs 1061170,-257C＞T rs3753394,IVS15 rs1329428),LOC387715/ARMS2 rs10490924 and HTRA1 rs11200638 play roles in the pathogenesis of exudative AMD and provide a theory basis of treatment and precaution of exudative AMD.