Proteoform profiling of peripheral blood serum proteins from pregnant women provides a molecular IUGR signature
Proteoform profiling of peripheral blood serum proteins from pregnant women provides a molecular IUGR signature
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DOI：
10.1016/j.jprot.2016.04.027
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发表时间：
2016-10-21
影响因子：
3.3
通讯作者：
Glocker，M. O.
中科院分区：
文献类型：
--
作者：
Woelter，M.;Roewer，C.;Glocker，M. O.
宫内生长受限（IUGR）是围产期发病率和死亡率的一个重要原因，并在很大程度上导致了医学上指征的早产；防止胎儿死亡。母亲外周血的分子谱分析是为了监测胎儿的健康状况。为了建立这样一种微创检测方法，我们对IUGR组和对照组孕妇的外周血血清样本采用了蛋白质亲和分离方法。通过使用MALDI-ToF质谱对完整的蛋白质形态混合物进行相对定量分析，证明了原理。载脂蛋白C-II和载脂蛋白c - ii0是两种最佳的分化蛋白和蛋白形态。它们与三种稳定表达的载脂蛋白C-II、载脂蛋白C-III1和载脂蛋白C-III2作为相对定量分析的标志蛋白一起构成了母体IUGR蛋白质组特征。我们的IUGR变形特征的分离置信灵敏度为0.73，特异性为0.87，接收算子特征的曲线下面积为0.86。由于儿童病情严重，需要在婴儿期进行专门护理，因此需要在病例室中识别IUGR新生儿。然而，在出生时，没有现成的适用的临床试验。因此，分子分析分析是非常需要的。需要指出的是，目前临床对IUGR的定义和建议不幸具有误导性，并不是普遍适用的。最常用的定义是腹围（AC）或估计胎儿体重测量值< 10个百分位数。尽管美国妇产科学院（ACOG）和英国皇家妇产科学院（RCOG）都认为，在这个临界值上，围产期发病率和死亡率的风险会增加，但这一定义并没有考虑到每个胎儿的个体生长潜力。特别是，它的唯一用途无法识别尚未达到其生长潜力并可能存在不良后果风险的较大胎儿。此外，如果单独使用这一定义，将导致对一些体质较小的胎儿的IUGR的误诊。需要指出的是，上述标准只有在母亲在怀孕早期报告的情况下才能在怀孕期间确定。我们开发了一种测试方法，它依赖于对母亲血清蛋白成分（IUGR特征）的质谱分析，可以在分娩前和分娩当日分别使用微创血液采样方法进行测定。在这篇文章中，我们描述了使用质谱分析方法，从孕妇分娩前的30外周血样本，无论是无可疑的新生儿或iugr影响的婴儿。我们首次报道了通过亲和质谱分析将IUGR婴儿与对照组区分开来的高可信度。
Intrauterine growth restriction (IUGR) is an important cause of perinatal morbidity and mortality and contributes substantially to medically indicated preterm birth; preventing fetal death. Molecular profiling of the mothers' peripheral blood was desired to monitor the health conditions of the fetuses. To develop such a minimally invasive assay, we applied a protein affinity fractionation method to peripheral blood serum samples from pregnant women belonging to either the IUGR or to the control group. Proof-of-principle was shown by relative quantitation analysis of mixtures of intact proteoforms using MALDI-ToF mass spectrometry. The two best differentiating proteins and proteoforms, respectively, were apolipoprotein C-II and apolipoprotein C-III0. Together with three robustly expressed protein proteoforms proapolipoprotein C-II, apolipoprotein C-III1, and apolipoprotein C-III2, which served as landmarks for relative quantitation analysis, they constituted the maternal IUGR proteome signature. Separation confidence of our IUGR proteoform signature reached a sensitivity of 0.73 and a specificity of 0.87 with an area under curve of 0.86 in receiver operator characteristics.SignificanceIdentification of IUGR newborns in the case room is required as children are severely diseased and need specialized care during infancy. Yet, at time of birth there is no readily applicable clinical test available. Hence, a molecular profiling assay is highly desired.It needs to be mentioned that current clinical definitions and recommendations for IUGR are unfortunately misleading and are not universally applicable. The most commonly adopted definition is an abdominal circumference (AC) or estimated fetal weight measurement < 10th percentile. Although both, the American College of Obstetricians and Gynecologists (ACOG) and the Royal College of Obstetricians and Gynecologists (RCOG) agree that at this cut-off the risk of perinatal morbidity and mortality increases, this definition does not take into account the individualized growth potential of each fetus. In particular its sole use fails to identify larger fetuses that have not achieved their growth potential and may be at risk of adverse outcomes. Also, this definition, when solely applied, will result in the misdiagnosis of IUGR for some constitutionally small fetuses. It needs to be pointed out that the above mentioned criteria can only be determined during pregnancy in case mothers report from early on during pregnancy.We have developed a test that relies on mass spectrometric analysis of the mother's serum protein composition (IUGR signature) which can be determined just ahead of delivery and at date of delivery, respectively using a minimal invasive blood sampling approach. With this manuscript we describe the use of a mass spectrometric profiling method of 30 peripheral blood samples from pregnant women prior to giving birth of either unsuspicious newborns or IUGR-affected infants.We report for the first time that maternal blood sample analysis via affinity mass spectrometry differentiates IUGR infants from controls with high confidence.