Associations Between The EHBP1-TUBB-WWOX SNPs And Serum Lipid Levels,Coronary Artery Disease And Ischemic Stroke

Object:With the development of life quality,hyperlipidemia has become a prevalent and silent murder for health.Genetic and various environmental factors elevate serum lipid levels leading to hyperlipidemia,which inducing obesity,hypertension,cardiovascular disease(CVD)and poor outcomes.Early prevention and treatment of hyperlipidemia is particularly important,and it is also a key point in the primary prevention of cardiovascular and cerebrovascular diseases.Previous studies revealed that there were differences in the expression of lipid-related genes between Han and Maonan populations,leading to different prevalence of hyperlipidemia and hypertension.GWAS found that single nucleotide polymorphisms(SNPs)rs2710642 and rs10496099 of EH domain binding protein 1 gene(EHBP1)were associated with serum low-density lipoprotein cholesterol(LDL-C)in European population.However,the associations between single nucleotide polymorphisms(SNPs)rs2710642 and rs10496099 and their effect on the EHBP1 and serum lipid profiles remain uncertain.This study was performed to investigate the two EHBP1 SNPs in Han and Maonan populations,including their association,haplotypes,and effects on serum lipid levels.It is to be a theoretical basis for early screening and prevention of hyperlipidemia in susceptible populations through predicting for optimal risk model of hyperlipidemia.Methods:It was a cross-sectional population cohort study.The Maonan and Han populations as the subjects recruited in Huanjiang region of Guangxi Zhuang Autonomous Region in 2015,and 564 Han and 796 Maonan samples were randomly selected using stratified random sampling(same ratio of gender and age)and divided into Han and Maonan groups.First,DNA was extracted,and the restriction fragment length polymorphism PCR method was employed to perform qualitative detection of EHBP1 rs2710642 on some samples,and then high-throughput sequencing was used to detect genotypes of EHBP1 rs271064and rs10496099 among all subjects.Finally,multiple linear regression and logistic regression were used to analyze the influence of the interaction of the two SNPs and their haplotypes with environmental factors on serum lipid levels in the two groups.Results:The total cholesterol(TC),LDL-C level,and apolipoprotein(Apo)A1/Apo B ratio were significantly higher in the Maonan than those in the Han population.On the contrary,the high-density lipoprotein cholesterol(HDL-C)and Apo B levels were significantly lower in the Maonan than those in the Han ethnic group.In Han and Maonan populations,a significant difference was found in the allelic and genotypic frequencies of the EHBP1 rs2710642 and rs10496099 SNPs.The genotypic(AA,GA and GG)and allelic(A and G)frequencies of SNP rs2710642 were 58.0%,38.0%,4.0%,77.0%and 23.0%in the Maonan group;were 41.0%,49.0%,10.0%,65.0%and 35.0%in the Han group.The genotypic(CC,TC and TT)and allelic(C and T)frequencies of SNP rs10496099 were 60.0%,36.0%,4.0%,78.0%and 22.0%in the Maonan group;were 50.0%,41.0%,9.0%,70.0%and 30.0%in the Han group.The alternate alleles of rs2710642A and rs10496099C might be potentially beneficial for healthy lipid levels.Medium linkage disequilibrium between the two SNPs was noted in each ethnic group,D’=0.820 or R~2=0.537 in the Han;D’=0.807 or R~2=0.620 in the Maonan;and four main haplotypes were detected(rs2710642G-rs10496099C,rs2710642A-rs10496099C,rs2710642G-rs10496099Tandrs2710642A-rs10496099T).The rs2710642G-rs10496099C haplotype was associated with high triglyceride(TG)and low HDL-C,and the rs2710642A-rs10496099C haplotype was associated with low TG and high Apo A1.The rs2710642G-rs10496099C haplotype was a high-risk factor for hyperlipidemia,and it interacted with smoking,fasting blood glucose,and hypertension to increase but with the female factor to decrease the prevalence of hyperlipidemia in Han individuals.Conclusions:There were associations between SNPs rs2710642,rs10496099,their haplotypes,and interactions of gene and environment on serum lipid levels and/or hyperlipidemia in Maonan and Han populations.The difference in serum lipid levels between the two groups might be contributed to the different expression in EHBP1 between the two ethnic groups,which might be ethnic specificity to our study populations.Object: The average global prevalence of dyslipidemia is about 20%,and it is higher in patients with premature coronary artery diseases and chronic kidney disease.In the past decades,technological advances in biological sequencing technology have enabled genome-wide association studies(GWASs),which help to thoroughly characterize the risk factors for dyslipidemia.GWASs found that tubulin beta class I(TUBB)and WW domain-containing oxidoreductase(WWOX)genes,were associated with serum low-density lipoprotein cholesterol(LDL-C)and/or high-density lipoprotein cholesterol(HDL-C)profiles.However,associations between TUBB and WWOX and lipid traits and dyslipidemia in the Chinese population have not been examined.We arm to explore the expressions of the four SNPs in Guangxi Maonan population,elaborated their genetic characteristics,and revealed associations between TUBB-WWOX SNPs,their haplotypes,and gene-gene(G × G)and gene-environment(G × E)interactions and the prevalence of dyslipidemia in the Maonan population.Methods: It was a cross-sectional study.A total of 1993 unrelated subjects were selected from our previous sample library in 2015 through stratified random sampling.Subjects were separated into normal lipid(864)and dyslipidemia(1129)groups with similar age and gender distributions.First,DNA was extracted,and then four SNPs(rs3132584,rs3130685,rs2222896,and rs2548861)were genotyped by high-throughput sequencing.Finally,binary logistic regression and generalized multifactor dimensionality reduction analysis were used to analyze the effects of G × G and G × E interactions on lipid levels and dyslipidemia in the two groups.Results: The lipid levels of TC,TG,HLD-C,LDL-C,Apo A1 and Apo B,and the ratio of Apo A1 / Apo B were significantly different in the two groups,as well as mean weight,waist circumference,body mass index(BMI),alcohol consumption,systolic blood pressure,diastolic blood pressure,pulse pressure,fasting blood-glucose(FBS),hypertension morbidity,and proportion with BMI greater than 24 kg/m2 and with FBS of at least 7.0 mmol/L were higher in the dyslipidemia group than in the normal group(P < 0.01).While 5.0% of Maonan subjects carried the rs3132584 TT genotype,none of the Chinese Han in Beijing subjects did.Allele and genotype frequencies differed between the normal and dyslipidemia groups for three SNPs(rs3132584,rs3130685 and rs2222896).rs2222896 G allele carriers in the normal group had higher low-density lipoprotein cholesterol and lower high-density lipoprotein cholesterol levels.Linkage disequilibrium was observed between the WWOX rs2222896 and rs2548861 SNPs in the normal and dyslipidemia groups(D′ = 0.870 or R2 =0.350 and D′ = 0.810 or R2 = 0.220,respectively).The rs3132584 GG,rs3130685CC+TT,and rs2222896 GG genotypes as well as the rs2222896G-rs2548861 G and rs2222896G-rs2548861 T haplotypes were associated with an elevated risk of dyslipidemia;the rs2222896A-rs2548861 T and rs2222896A-rs2548861 G haplotypes were associated with a reduced risk of dyslipidemia.Among the thirteen TUBB-WWOX interaction types identified,rs3132584T-rs3130685T-rs2222896G-rs2548861 T increased the risk of dyslipidemia 1.371-fold.Fourteen two-to four-locus optimal interactive models for SNP-SNP,haplotype-haplotype,G × G,and G × E interactions exhibited synergistic or contrasting effects on dyslipidemia.Finally,the interaction between rs3132584 and rs2222896 increased the risk of dyslipidemia 2.548-fold and the risk of hypertension 1.523-fold.Conclusions: This study identified associations between TUBB,WWOX,environmental factors,serum lipid levels,and dyslipidemia in the Maonan population.Our findings revealed that,compared to single-locus effects,TUBB-WWOX-environment interactions can result in synergistic or contrasting effects on serum lipid levels,thereby increasing or reducing the risk of dyslipidemia.Object: Recently,the treatment of hyperlipidemia,hypertension and diabetes is still the key to the primary prevention of cardiovascular and cerebrovascular diseases.Statin is the traditional drug for lipid-lowering,in the meanwhile,gene therapy has been used for refractory hyperlipidemia.Our previous research found that associations between EHBP1,TUBB and WWOX SNPs and serum lipid levels and dyslipidemia in Guangxi Han and Maonan populations,revealing that dyslipidemia of the two populations was affected by genetic and regional environmental factors.In view of the close correlation between dyslipidemia and atherosclerotic diseases,we speculate that there may be associations between the three genes and coronary artery disease(CAD)and ischemic stroke(IS)in Guangxi Han population.We arm to investigate the expression levels of EHBP1 rs2710642 and rs10496099,TUBB rs3132584 and rs3130685,and WWOX rs2222896 and rs2278075,to elaborate genetic characteristics of the six SNPs,and to reveal associations between SNPs,their haplotypes,EHBP-TUBB-WWOX and gene-environment(G × E)interactions and CAD and IS in the Han.Screening the optimal predict model for disease risk,it might provide a basic theory of new target treatments for primary prevention for dyslipidemia in patients with CAD and IS,and ultimately achieve the goal of reducing the prevalence of ischemic cardio-cerebrovascular disease.Methods: It was a cross-sectional and case-control study.A total of 1853 unrelated subjects,who were physical examination,outpatient visits or hospitalization in the First Affiliated Hospital of Guangxi Medical University from September 2009 to December 2011,were divided into normal control(n=638),CAD(n = 622),and IS(n = 593)groups.Identifying the HWE conditions satisfied,we analyzed the correlation between these SNPs,their haplotypes and CAD and IS and blood lipid levels,and then explored the associations between G × G,G × E and CAD and IS,and finally screened out the optimal forecast models for risks of CAD and IS.Results: The mean height,systolic blood pressure,diastolic blood pressure,pulse pressure,daily alcohol and cigarette consumption,and the proportion with antihypertensive treatment of the CAD or IS,were significantly different from those of the control.The prevalence of hypertension in the IS,and the hypoglycemic rate in the CAD were higher than those in the control,respectively(P < 0.001).The genotypic and minor allelic frequencies of rs2278075 were statistically different between the CAD and control groups,and those of rs2710642,rs3130685,rs2222896 and rs2278075 were different between the IS and control groups.Linkage disequilibrium(LD)was observed between the EHBP1 rs2710642 and rs10496099 SNPs in the three groups.In the control,CAD and IS,D’ = 0.951 or R2 = 0.821,D’ = 1.000 or R2 = 0.938,D’ =1.000 or R2 = 0.690,respectively;while there was no LD among other SNPs in each group.The rs2278075 T allele,rs3130685-rs2222896-rs2278075,rs3130685-rs2222896-DM,and rs3130685-rs2222896-drinking interactions increased while I5-I6-drinking reduced the risk of CAD.The rs2278075 T and rs2710642 G alleles,rs2710642G-rs10496099 C haplotype and rs3130685-rs2278075-rs2222896,rs2710642-rs2278075-hypertension interactions aggravated the risk of IS;rs3130685T or rs2222896 A allele,rs2710642A-rs10496099 C haplotype,the interaction of haplotype rs2710642A-rs10496099 C and haplotype rs2710642G-rs10496099C-drinking,and the interaction of A-C-G-C-A-A and A-C-G-T-A-A and A-C-G-T-G-A and G-T-G-C-G-A declined the risk of IS.There was no difference in the average levels of TC,LDL-C,Apo B and the rate of lipid-lowing treatment between the control group and CAD group or the IS group(P ≥ 0.05).However,the levels of TG in the CAD or IS groups were higher than those in the control group(P ≤0.003),and the levels of HDL-C and Apo A1,and the ratio of Apo A1 / Apo B were significantly lower than those of the control group(P < 0.001).The rs2278075 T was associated with low level of LDL-C and high level of Apo A1 in CAD group.The rs2278075 T allele and rs2710642A-rs10496099 C haplotype were associated with low levels of TC,TG and LDL-C and high level of HDL-C;rs2710642G and rs3130685 T alleles were correlated with high levels of TC,TG and LDL-C;and rs2222896 G allele was correlated with high levels of LDL-C and TG in IS group.Conclusions: WWOX rs2278075 T carried led to CAD or IS;while EHBP1rs2710642,TUBB rs3130685 and WWOX rs2222896 might alter the risk for IS through modifying blood lipid profile.The haplotypes of EHBP1 rs2710642 and rs10496099 were more valuable in predicting IS.Associations between these interactions of gene and gene,interactions of genes and environmental factors including women,drinking,smoking,diabetes mellitus and hypertension,determined the risk of CAD and IS in Guangxi Han population.