Research On Proteomics Of Preeclampsia Based On Mass Spectrometry

Background and purposePreeclampsia(PE)is a common hypertensive disease in pregnancy,which seriously affects maternal and infant morbidity and mortality worldwide.PE is a multi-system disease in which various etiologies affect each other.Therefore,specific monitoring indicators and methods for early diagnosis or prediction of disease severity are lacking.Proteomics studies based on mass spectrometry provide essential breakthroughs in understanding the pathophysiology of complex human diseases by discovering biomarkers.In this project,DIA-PRM technology platform was used to carry out proteomics analysis on blood and urine samples of preeclampsia.By comparing the results of prenatal and postnatal differential proteins from normal pregnancy and preeclampsia patients,we screened differential proteins with high specificity and sensitivity for identification as biomarkers.MethodsThe blood and urine samples of 7 normal pregnancies and 5 preeclampsia pregnant women were collected 24 hours before and after delivery.The differential proteins were screened by quantitative proteomics method of data independent acquisition(DIA),and analysised by ingenuity pathway analysis(IPA).We collected urine samples 24 hours before and after delivery from 20 normal pregnancies,25 cases of preeclampsia,and 15 cases of gestational hypertension.Parallel response monitoring(PRM)technique was used to verify the targeted differential proteins.ROC curves and AUC were constructed for the verified differential proteins to search for biomarkers with high identification efficiency.ResultsIn the first part,274 differential proteins were screened out in serum by DIA technique,including 84 in the preeclampsia group and 190 in the control group.There were 671 differential proteins in urine,including 449 in preeclampsia group and 222 in control group.Differential proteins were up-regulated and down-regulated in different degrees,respectively.Bioinformatics analysis showed that differential proteins in preeclampsia were abnormal in inflammatory immune response,endothelial injury,cell proliferation and angiogenesis,and significantly positively activated in renin angiotensin signaling pathway,adrenomedullin signaling pathway and myocardial hypertrophy signaling pathway.Important functional differential proteins in the serum and urine of preeclampsia and normal pregnancy have a consistent trend,however,in the preeclampsia group,the expression of PRDX2,CD99 and ITGB1 proteins was down-regulated in serum and upregulated in urine after delivery,which was inconsistent with the normal group.In the second part,urine samples were selected from three groups of preeclampsia(n=20),normal pregnancy(n=18)and gestational hypertension(n=11).A total of 109 proteins(157 polypeptides)were quantified by PRM,and 73 proteins(99 peptides)were consistent with DIA trend,among which 28 proteins(35 peptides)were up-regulated before delivery and 45 proteins(64 peptides)were down-regulated before delivery.In line with the DIA trend(p<0.05)showed 16 proteins(18 peptide),of which 2 proteins(2 peptide)were up-regulated and 14 proteins(16 peptide)were down-regulated.Combined with scatter plot and ROC curve,11 candidate biomarkers with differential significance were screened out,namely MXRA5,VGF,VATA,NTF2,LTBP2,FLNA,FAT4,CREL1 LIRB4,FIBB,PRDX2,which can be used as candidate biomarkers to distinguish preeclampsia from normal pregnancy.Among them,MXRA5,VGF,VATA,NTF2,LTBP2,FLNA can be used as candidate biomarker protein to distinguish preeclampsia,normal pregnancy and gestational hypertension.The identification efficiency of protein VGF was the highest,with AUC=0.939,and the combination of three proteins VGF,CREL1 and LTBP2 with AUC=0.95(95%CI:0.854-1)had a higher differentiation efficiency between preeclampsia and normal pregnancy.The combination of FLNA and LTBP2 with AUC=0.92(95%CI:0.5-1)showed high efficiency in differentiating preeclampsia from gestational hypertension.Conclusions1.Differential proteins in ferroptosis signaling pathway were significantly activated,which may be associated with preeclampsia related injury.2.Eleven differential proteins MXRA5,VGF,VATA,NTF2,LTBP2,FLNA,FAT4,CREL1,LIRB4,FIBB and PRDX2 may serve as candidate biomarkers to distinguish preeclampsia from normal pregnancy.Among them,the combination of three differential proteins VGF,CREL1 and LTBP2(AUC=0.95)has higher value in distinguishing preeclampsia from normal pregnancy.3.Six differential proteins MXRA5,VGF,VATA,NTF2,LTBP2 and FLNA may be used as candidate biomarkers to distinguish preeclampsia,normal pregnancy and gestational hypertension.Among them,the combination of FLNA and LTBP2(AUC=0.92)has higher value in differentiating preeclampsia from gestational hypertension.