| Part One Association between ATG5 Gene Polymorphisms and Anti-neutrophil Cytoplasmic Antibody Associated VasculitisObjective: The significance of ATG5 single nucleotide polymorphisms in primary anti-neutrophil cytoplasmic antibody(ANCA)associated vasculitis(AAV)has not been studied.The purpose of this study was to investigate the association between 10 single nucleotide polymorphism(SNP)in ATG5 gene and ANCA-associated vasculitis in the Han and the Zhuang population in Guangxi.Methods: 177 patients(AAV group)who met the inclusion criteria of AAV and216 healthy people(control group)whose degree of age,sex was similar to AAV group from the Han and Zhuang population in Guangxi were collected for the study.The control group included 158 Han and 58 Zhuang ethnic people.The AAV group included 115 cases of Han and 62 cases of Zhuang.Clinical data of the subjects were collected and DNA was extracted from whole blood.Genotyping of selected SNPs was performed using multiple polymerase chain reaction combined with high throughput sequencing.A case-control study was conducted on the Han and Zhuang populations respectively to analyze the distribution differences of SNP genotypes,alleles and their common haplotypes in the control group and the AAV group.Results:(1)The 10 SNPs of ATG5 gene included in the study were rs617994,rs1766193,rs656994,rs148316514,rs573775,rs662114,rs510432,rs506027,rs3761796 and rs473543,which were all present in the Han and Zhuang populations in Guangxi.(2)There was no significant difference in the genotype frequency and allele frequency between the control group and the AAV group(P>0.05).(3)In the Han and Zhuang populations of Guangxi,there were 6 common haplotypes made by 10 SNPS of ATG5 gene.In the Han population in Guangxi,haplotype VI was associated a lower risk of AAV compared to haplotype I(OR =0.24,95% CI=0.07-0.84,P=0.027).Conclusion:(1)The ATG5 genes rs617994,rs1766193,rs656994,rs14831654,rs573775,rs662114,rs510432,rs506027,rs3761796,rs473543 are poly morphic in the Han and Zhuang populations of Guangxi,but they may ha ve nothing to do with the genetic susceptibility of AAV in the Zhuang p opulation of Guangxi.(2)In the Han population the haplotype rs617994G-rs1766193T-rs656994T-rs14831654C-rs573775A-rs506027A-rs473543 G may ha ve an influence on AAV.Part Two Association between ATG16L1 Gene Polymorphisms and Anti-neutrophil Cytoplasmic Antibody Associated VasculitisObjective: the purpose of this study was to investigate the relationship between 10 single nucleotide polymorphisms at ATG16L1 gene and anca-associated vasculitis in Han and Zhuang populations of Guangxi.Methods: genotyping of selected SNPS was performed using multiple polymerase chain reaction(PCR)combined with high-throughput sequencing,and a case-control study was conducted in 273 Han and 120 Zhuang populations.Results:(1)The 10 SNPS of ATG16L1 gene included in the study were rs10210302,rs6737398,rs2241880,rs3792109,rs2241877,rs3792106,rs4663396,rs4663402,rs4663421 and rs76072155,and all alleles were detected in Han and Zhuang populations(frequency greater than 0.01).(2)All of alleles of 10 SNPs of ATG16L1 gene were found in the Han and Zhuang populations in Guangxi,however,there was no significant difference in genotype frequency and allele frequency between the control group and the AAV group.(3)rs10210302,rs6737398,rs2241880,rs3792109 and rs2241877 are all strong linkage disequilibrium in the Han and Zhuang populations,constituting four common haplotypes which were not associated with AAV.Conclusion:(1)It showed polymorphisms of rs10210302,rs6737398,rs2241880,rs3792109,rs2241877,rs3792106,rs4663396,rs46634402,rs4663421,rs76072155 in ATG16L1 gene in the Han and Zhuang populations of Guangxi.(2)rs10210302,rs6737398,rs2241880,rs3792109,rs2241877,rs3792106,rs4663396,rs4663402,rs4663421,rs76072155 may not be associated with the AAV in Han and Zhuang populations of Guangxi.Part Three Studies on the Association between IRGM Gene Polymorphisms and ANCA Associated Vasculitis and the Relationship between Genotypes and IRGM Gene ExpressionObjective: the purpose of this study was to investigate the association between single nucleotide polymorphisms at 5 sites of IRGM gene and ANCA-associated vasculitis in Han and Zhuang populations of Guangxi.Methods: genotyping of selected SNPS was performed using multiple polymerase chain reaction(PCR)combined with high-throughput sequencing,and a case-control study was conducted in 273 Han and 120 Zhuang populations.The relationship between genotypes and gene expression of SNPs which is associated with AAV was preliminarily explored using GTEx database,this result was verified by real-time quantitative PCR in AAV patients from Zhuang population in Guangxi.Results:(1)The five SNPs of the IRGM gene included in the study were rs13361189,rs72553867,rs10065172,rs1000113 and rs11743488,which were present in both the Han and Zhuang populations of Guangxi.However,rs13361189,rs10065172,rs1000113 and rs11743488 did not meet the Hardy-Weinberg equilibrium in the Guangxi Han population included in the study,and further analysis was not performed.(2)In the Zhuang ethnic group of Guangxi,the differences in genotype frequency and allele frequency of rs13361189,rs10065172,rs1000113 and rs11743488 between the control group and the AAV group were statistically significant(all P< 0.05).Further analysis showed that the rs13361189 C allele was associated with lower risk of AAV(Recessive: OR=0.39,95%CI=0.15-0.99,P=0.043;Log-additive: OR=0.50,95%CI=0.29-0.86,P=0.010).The rs10065172 T allele was associated with lower risk of AAV(Recessive: OR=0.39,95%CI=0.15-0.99,P=0.043;Log-additive: OR=0.50,95%CI=0.29-0.86,P=0.010).The rs1000113 T allele was associated with lower risk of AAV(Recessive: OR=0.36,95%CI=0.14-0.91,P=0.026;Log-additive: OR=0.51,95%CI=0.30-0.87,P=0.011).The rs11743488 G allele was associated with lower risk of AAV((Recessive: OR=0.28,95%CI=0.10-0.78,P=0.010;Logadditive: OR=0.46,95%CI=0.26-0.80,P=0.004).(3)rs13361189,rs10065172,rs1000113,rs11743488 present strong linkage disequilibrium,rs13361189 was complete linkage disequilibrium with rs10065172,4 SNPs constitute four kinds of common haplotype,overall haplotype was associated with risk of AAV risk(P = 0.023),haplotype II was associated with a decreased risk of AAV compared to haplotype I(OR = 0.47,95% CI = 0.27 0.82,P=0.009).(4)The result of e QTL analysis using data from GTEx database and showed that there was a difference in between rs13361189 and rs10065172 genotypes and IRGM expression,and was verified by real-time quantitative PCR in AAV patients from Guangxi Zhuang population.Conclusion:(1)It showed polymorphisms of rs13361189,rs72553867,rs10065172,rs1000113 and rs11743488 in the Han and Zhuang populations of Guangxi.(2)In the Zhuang population of Guangxi,rs13361189,rs10065172,rs1000113,rs11743488 and their haplotypes may be associated with the risk of AAV.(3)There were differences in IRGM expression between the genotypes of rs13361189 and rs10065172 in AAV patients from Guangxi Zhuang population. |
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