High-throughput Technology Is Used To Screen The Genes Associated With Longevity And Ankylosing Spondylitis

Study on healthy longevity phenotype and genotype in south of ChinaAim Lifespan would be extending with the improve of health care level.Most country must face the problem of aging population more or less.Longevity is a complex trait,and its formation is the result of many factors.Recent study had showed that genetic factors played an important role in longevity.Searching longevity associated gene or variants could help us understanding causes of longevity,reduce the occurrence of senile diseases,protect the health of the elderly,and promote the healthy aging of society.Methods All samples were collected in Guangxi,Bama and Yongfu,and CLHLS.There 156 long-lived individuals and 116 controls in Bama population,482 long-lived individuals and 404 controls in Yongfu population.CLHLS database contains 362 long-lived individuals and 115 controls.100 whole-exomes were sequenced by Illumina 2500,containing 32 males and 68 females.Longevity associated candidate variants were selected by comparing sequencing result to 1000 Genome CHS variants database,and replicated in Bama sample set and CLHLS database.Previously reported longevity-associated variants were analyzed in Bama sample set and CLHLS database.Allele of disease,type Ⅱ diabetes mellitus,coronary disease,stroke,cancer,Alzheimer’s disease,distribution comparison was proceeded between Bama long-lived individuals and CHS.Candidate longevity-associated low-frequency variants were selected by frequency differences,and these variants were genotyped in Bama and Yongfu sample set,using PCR-HRM,PCR-RFLP and Sequenom MassARRAY iPLEX.All these data was summaried to identify longevity-associated variatns.The genetic effect on longevity related phenotypes was also analyzed.Results MUC4 rs199822551,MUC4 rs79358488,MUC4 rs71635077,MUC4 rs201141296,MUC4 rs72499650,PRIM2 rs62415470,MUC3A rs77036391,MUC3A rs78164286,MUC6 rs34844844,MUC19 rs2452315,PARP4 rs201405094,AHNAK2 rs2582506 and MAP2K3 rs55935757 were detedted as longevity-associated variatns by comparing Bama sequencing sample set and CHS population.Meta-analysis was proceeded in Bama sample set and CLHLS database,and MAP2K3 rs55935757(P＝.031 × 10-7,OR:1.755,95%CI:1.427-2.159,I2＜0.001)was detedted as a longevity-associated variant.The association between APOE rs440446 and longevity had been verificated repeatedly(P=0.030,OR:1.251,95%CI:1.022-1.531,I2:64.82).There was no significat association of allele distribution,what was associated with T2DM,CAD,stroke,cancer or AD,between longevity and CHS population.TAS1R2 rs200812417,TTN rs4893852,TTN rs13398235,APOB rsl3306187,WNT1OA rsl47680216,ANKRD28 rs77003664,TMEM156 rs2276887，FAM13A rs41284761,PDHA2 rs35423663,PRDM5 rs146228268，ABHD18 rs560511632,CEP44 rs150156902,ERAP1 rs27985,ASPHrsl 83457686,EPB41L4B rs149000673,DAB2IP rs202232151,HIFNT rs2732441,ZFN641 rs2634680,ZFN641 rs2732481,ATP5G2 rs17101603,RTNI rs201516762,GALNS rs2303269 and MPP3 rs 17742683 were detected as longevity associated variants in Bama population.ZFN641 rs2732481 and ATP5G2 rs17101603 were verificated in Yongfu population.Meta-analysis result showed TTN rs4893852,TTN rs13398235,TMEM156 rs2276887,PDHA2 rs35423663,ABHD18 rs560511632,EPB41L4B rS149000673,ZFN641 rs2732481,ATP5G2 rs17101603 and RTN1 rs201516762 were associated with longevityConclusions MAP2K3 rs55935757 C/T was associated with longevity in Chinese.The association between APOE rs440446 C/G and longevity was verificated.Longevity population carries less disease risk allele than common population.Low frequency variants,includes TTN rs4893852,TTN rs 13398235,TMEM156 rs2276887,PDHA2 rs35423663,ABHD18 rs560511632,EPB41L4B rs149000673,ZFN641 rs2732481,ATP5G2 rs17101603 and RTN1 rs201516762,were associated with longevity in Guangxi population.Identification of Ankylosing Spondylitis associated low frequency variantsAim Ankylosing spondylitis(AS)is a chronic inflammatory arthritis that affects the spine and sacroiliac joints.AS is a polygenic disease.Recent study showed AS pathogeny was associated with genetics,infections and environmental factors.Multiple GWAS results showed HLA region in chromosome 6 was the strongest associated loci with AS.HLA-B27 is a classic AS-associated locus.Then previous studies suggested that there might be association between HLA Class III region and AS.We had mapped the underlying susceptible loci to AS within a 58-kb segment ranging from BAT1 to NCR3 in the HLA Class III region.With the progress of study,we had identified two new AS associated loci,TNFa rs 1799724 and LTA rs909253.We hypothesized that there might be unknown AS associated genes.This study was preformed to investigate and verify the association of low frequency variations in HLA Class III region and ankylosing spondylitis susceptibility in Northern Chinese populationsMethods 901 ankylosing spondylitis patients and 956 healthy controls were collected in this study.There were 136 patients and 148 controls in Jilin population,269 patients and 257 controls in Beijing population,496 patients and 551 controls in Ningxia population.rs13202464,rs4418214,HCP5 rs192776040,AIF1 rs369457101,BAG6 rs201165258,C2 rs574394257,SK1V2L rs144147284 and TNXB rs141190850 were genotyped in ankylosing spondylitis cases and healthy controls,using high resolution melting.The association of variants and ankylosing was analyzed in the combined population which was formed by Jilin,Beijing and Ningxia population,and the relationship of ankylong spondylitis associated low frequency variants in HLA Class III and HLA-B27.Results Significant difference was detected in genotype and allele distribution of rs 13202464,rs4418214,HCP5 rs 192776040,AIF1 rs369457101,BAG6 rs201165258and C2 rs574394257 in Jilin,Beijing and Ningxia sample sets(P<0.05).Dominant models of 8 variants were all associated with AS(P＜0.05).Binary logistic regression result showed,rs13202464 was associated with AS β=1.797,P=5.923 X 10-36,ORadj:6.029,95%CI:4.551-7.988);rs4418214 was associated with AS(β=1.764,P=5.000 × 10-30,ORadj:5.833,95%CI:4.306-7.903);HCP5 rs192776040 was associated with AS(3=3.488,P=4.675 X 10-33,ORdj:32.708,95%CI:18.484-57.877);C2 rs574394257 was associated with AS(β=0.752,P=0.030,ORadj:2.121,95%CI:1.074-4.191);TNXB rs141190850 was associated with AS(β=1.841，P=0.001，ORadj:6.301,95%CI:2.133-18.610).Linkage disequilibrium analysis result showed there were two Block in this region,Block 1 formed by AIF1 rs369457101,BAG6 rs201165258 and C2 rs574394257,Block 2 formed by SKIV2L rs144147284 and TNXB rs141190850.Haplotype analysis results showed there were two haplotypes in Block 1,CAT and TGG and Block contained two haplotypes,GA and AG.In Block 1,the distribution of CAT haplotype in cases and controls was significant(P=1.015×10’29,OR:0.157,95%CI:0.109-0.226).The distribution of TGG haplotype in cases and controls was significant(P=2.889×10-25,OR:14.064,95%CI:7.359-26.878).In Block 2,the distribution of GA haplotype in cases and controls was significant(P=2.763×10’16,OR:0.060,95%CI:0.024-0.150).the distribution of AG haplotype in cases and controls was significant(P=1.939X 10-12,OR:13.894,95%CI:5.011-38.523).In HLA-B27 positive group,carrying rs4418214 G allele,HCP5 rs192776040 T allele,AIF1 rs369457101 T allele,BAG6 rs201165258 G allele or C2 rs574394257 G allele could increase AS risk(P＜0.05).In HLA-B27 negative group,carrying rs4418214 G allele,HCP5 rs192776040 T allele,AIF1 rs369457101 T allele,BAG6 rs201165258 G allele,C2 rs574394257 G allele or TNXB rs141190850 A allele could increase AS risk(P<0.05).Conclusions HCP5 rs192776040,AIF1 rs369457101,BAG6 rs201165258,C2 rs574394257,SKIV2L rsl44147284 and TNXB rs141190850 would associated with ankylosing spondylitis,independent with HLA-B27 in Chinese northern population.Carrying minor alleles of these variants could increase the risk on ankylosing spondylitis.