Association Between Sex Hormone Metabolism And Angiogenic Pathway Genes Polymorphism With Preeclampsia

Objective:Although aberrant synthesis and metabolism of sex-hormone are likely to be associated with alterations in vascular function in preeclampsia,epidemiologic studies have not found clear associations between preeclampsia and sex-hormone metabolism.The study aims to investigate whether single nucleotide polymorphisms?SNPs?in sex hormone related genes are associated with preeclampsia.It may be one of importance role in the underlying mechanism of preeclampsia that remodeling of the maternal spiral arteries reduced.The aim of the study is to implore that single nucleotide polymorphisms?SNPs?in angiogenic related genes are associated with preeclampsia.Expreimental data have revealed that the sex hormone could reduce angiogenesis dysfunction in the maternal blood with preeclampsia,and these studies may provide clues about preeclampsia etiology.We therefore invested the relationships between sex hormone and angiogenic related genes polymorphisms and the risk of preecampsia.Method:Objectives and Research methodology:We conducted a nested case-control analysis based on 203 women with preeclampsia as case group and 233 normal controls,the patients were recruited for the study at the Department of Obstetrics and Gynecology,the First Affiliated Hospital of Shanxi Medical University in Taiyuan,China between March 2012 and November 2013.203 cases selection was based on diagnostic criteria of gestational hypertension disease diagnosis and treatment.We had been randomly selected 233 controls from the same population and being frequency matched to cases on year of birth?±2 years?,residence,and gestation weeks?±3 months?.The patients gave singleton live birth without major birth defects,and had no chronic hypertension and cardiovascular diseases.An in-person interview was conducted using a standard questionnaire to collect information on demographic factors?past medical history,passive/initiative smoking,drinking?,reproductive and medical history.The majority of clinical data?information on birth outcomes and pregnancy complications?was abstracted from medical database.All the blood samples were drawn at around the time of delivery.Experimental methods:DNA was extracted,isolated and purified from whole blood samples by DNA extraction Kit?Qiagen,USA?.Genotyping was conducted using an Illumina Golden gate Platform.A total of 111 SNPs from 28 sex hormones pathway genes and 15 SNPs from 6 angiogenic pathway genes were considered for this study.For quality control,about 5%random selected duplicates were included.Statistical approachs:Statistical analyses were performed using the SAS software version 9.4?SAS Institute?.We analyzed the basic characteristics of case and control by Chi-square test and t-test.We examined an association at the gene level through the Minimum P?“MinP”?tests.All study SNPs conforms to the Hardy-Weinberg Equilibrium in the genetal population by Chi-square test.Unconditional logistic regression models were used to explore the associations between two pathways genes polymorphism and preeclampsia and its subtypes,and calculate odds ratios?ORs?and95%confidence intervals?CIs?for individual SNPs adjusting for age,education level,and body mass index.Haplotype analyses were conducted for all genes in which more than one SNP was genotyped.Haplotype block structures were evaluated with HaploView using the four gamete rule.We estimated the individual haplotype frequencies through the expectation maximization algorithm under the assumption of the Hardy-Weinberg Equilibrium.An unconditional logistic regression model was used to estimate the effect of individual haplotypes,with the most common haplotype as the reference.As we were testing different SNPs,we reported q-values,a measure of significance in terms of the false-discovery rate?FDR?as proposed for multiple comparisons.We considered a q-value less than 20%to be significant.Results:A nested case-control study was conducted,the equilibrium test between case and control group was comoparable.In the present study,we examined sex hormone metabolism and angiogenesis pathway genes polymorphism and explored the association between SNP and PE.There were 28 genes?94 SNPs?and 6 genes?12SNPs?in these two pathways.1.Associations between Sex hormone pathway genes polymorphism and the risk of preeclampsia were showed.Sex hormone biosynthesis and metabolism and Gonadotropin and germ cell development genes:TXNRD2/COMTrs3788314(OR_GG=2.15,OR_AG&GG=1.68),SULT1A2/SULT1A1rs4788073(OR_AG=1.67,OR_GG=4.01,OR_AG&GG=1.82)and CYP17A1rs4919690(OR_AG=1.65,OR_AG&GG=1.66),rs4919687(OR_AG=1.50,OR_AG&GG=1.54),LHCGRrs10180731(OR_AC&GG=1.45)were associated with an increased risk of preeclampsia overall.HSD17B3 rs8190512(OR_AC=0.61,OR_CC=0.55,OR_AC&CC=0.59)were associated with a decreased risk of preeclampsia overall.There were difference associations between SNPs and preeclampsia's subtypes.TXNRD2/COMTrs3788314,SULT1A2/SULT1A1rs4788073,HSD17B3rs8190512 and LHCGRrs10180731 were associated with early-onset and severe preeclampsia.CYP17A1rs4919690 and rs4919687 were associated with an increased risk of late-onset and severe preeclampsia,while GNRHRrs2630488 was associated with a decreased risk of severe preeclampsia(OR_AG=0.57,OR_AG&GG=0.56).We also observed that SULT1A2/SULT1A1rs4788073 was associated with an increased risk of mild preeclampsia.All SNPs,with related to preeclampsia,were predicted how exonic mutations disrupt the correct pattern of pre-mRNA splicing by ESEfinder3.0software.Significant difference in haplotype frequencies of HSD17B3,LHCGR,SULF1 was also found between preeclampsia and the healthy controls.The haplotype block?HSD17B3 rs10739847&rs11788785&rs13302476&rs8190512?was associated with the risk of preeclampsia,and an increased risk of preeclampsia was observed in A-A-A-A and C-A-A-C genotype of the block compared to reference.However,a reduced risk of preeclampsia was observed in C-A-A-C and A-C-C genotype of the block?LHCGRrs12469065&rs10180731&rs4594496&rs4245821,SULF1rs10106958&rs7813987&rs6990375?compared to reference.2.Associations between angiogenesis pathway genes polymorphism and the risk of preeclampsia were showed,but none of the SNPs remained significant after correction for multiple comparisons in preeclampsia overall.While we observed that FLT-1rs12584067 was associated with an increased risk of early-onset preeclampsia(OR_GC=1.98,OR_GC&GG=1.99).Additionally,VEGFArs3025039 was associated with an increased risk of mild preeclampsia(OR_TT=3.14)and ENGrs10121110 was associated with an increased risk of severe preeclampsia(OR_TC=2.17).We also observed that VEGFArs3025039(OR_TC&TT=0.50),VEGFCrs1485766(OR_AC=0.50,OR_AC&CC=0.50)and VEGFArs699947(OR_AC&AA=0.55)were associated with a decreased risk of severe preeclampsia.All non-coding genes polymorphism,with related to preeclampsia,were predicted how exonic mutations disrupt the correct pattern of pre-mRNA splicing by ESEfinder 3.0 software.3.Associations between sex-hormone-metabolism-angiogenesis pathway genes polymorphism and the risk of preeclampsia were showed.TXNRD2/COMT rs3788314(OR_AG=2.53,OR_AG&GG=1.65),SULT1A2/1A1rs4788073(OR_AG&GG=1.71)and LHCGRrs10180731(OR_GG=2.29,OR_AG&GG=1.53)were associated with an increased risk of preeclampsia overall.HSD17B3rs8190512(OR_AC&CC=0.46)and VEGFCrs1485766(OR_AC=0.59,OR_CC=0.50,OR_AC&CC=0.60)was associated with aR decreased risk of preeclampsia overall.TXNRD2/COMTrs3788314,ENG rs10121110 and VEGFCrs1485766 were associated with early-onset preeclampsia.TXNRD2/COMTrs3788314,SULT1A2/1A1rs4788073,CYP17A1rs4919690,LHCGRrs10180731 and HSD17B3rs8190512 were associated with early-onset preeclampsia.TXNRD2/COMTrs3788314,SULT1A2/1A1rs4788073,LHCGR rs10180731,VEGFArs3025039 and GNRHRrs2630488 were associated with mild preeclampsia.TXNRD2/COMTrs3788314,SULT1A2/1A1rs4788073,LHCGR rs10180731,ENGrs10121110,VEGFArs3025039,rs699947 and VEGFCrs1485766were associated with severe preeclampsia.Conclusions:Based on these results,SNPs in selected hormone pathway genes appear to be strongly related to preeclampsia and its subtypes.Our study suggested that genetic polymorphisms in TXNRD2/COMTrs3788314,SULT1A2/SULT1A1rs4788073,CYP17A1rs4919690 and rs4919687,HSD17B3rs8190512,GNRHRrs2630488,LHCGRrs10180731 might play a role in preeclampsia risk,especially in late-onset preeclampsiaandseverepreeclampsia.GenesofTXNRD2/COMT,SULT1A2/SULT1A1,CYP17A1,HSD17B3 might play a key role in sex hormone metabolism,and gene of GNRHR,LHCGR might play a role in signal pathway of sex hormone functions.Our results suggest that SNPs in selected angiogenesis pathway genes appear to be strongly related to subtypes preecampsia.The five SNPs in the VEFGA rs3025039,rs699927,FLT-1 rs12584067,VEGFC rs1485766,ENGrs10121110 may be associated with preeclampsia's subtypes,especially early-onset and severe preeclampsia.Our study provided that genetic polymorphisms in TXNRD2/COMTrs3788314,SULT1A2/1A1rs4788073,CYP17A1rs4919690,HSD17B3rs8190512,LHCGR rs10180731,GNRHRrs2630488,VEGFArs699947,rs3025039,VEGFCrs1485766,ENGrs10121110 were associated with the preeclampsia pathogenesis and the risk varied by preeclampsia subtypes.The significant associations observed for preeclampsia overall were mainly seen for late-onset preeclampsia and severe preeclampsia.We suggested that these gene polymorphisms may affect the expression and function of proteins.The two pathways sex-hormone metabolism and angionesis might be mutual regulatory relationship in the pathogenesis of preeclampsia and its subtypes.Future studies are warranted to replicate the observed associations and their functional mechanisms.