Divisions of Maternal Fetal Medicine and Reproductive Genetics, Department of Obstetrics and Gynecology and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA 15213, USA.
Placenta. 2011 Jan;32(1):70-8. doi: 10.1016/j.placenta.2010.10.004. Epub 2010 Nov 13.
Preeclampsia is a major obstetrical complication affecting maternal and fetal health. While it is clear that there is a substantial placental contribution to preeclampsia pathogenesis, the maternal contribution is less well characterized. We therefore performed a genome-wide transcriptome analysis to explore disease-associated changes in maternal gene expression patterns in peripheral blood mononuclear cells (PBMCs).
Preeclampsia was defined as gestational hypertension, proteinuria and hyperurecimia. Total RNA was isolated from PBMCs obtained from women with uncomplicated pregnancies (n = 5) and women with preeclamptic pregnancies (n = 5). Gene expression analysis was carried out using Agilent oligonucleotide microarrays. Biological pathway analysis was undertaken using Ingenuity Pathway Analysis software. Quantitative real-time PCR (QRTPCR) was performed to validate the gene expression changes of selected genes in normotensive and preeclamptic patients (n = 12 each).
We identified a total of 368 genes that were differentially expressed in women with preeclampsia compared to normal controls with false discovery rate (FDR) controlled at 10%. In follow up experiments we further analyzed the expression levels of a number of genes that were identified as altered by the microarray data including survivin (BIRC5), caveolin (CAV1), GATA binding protein-1 (GATA1), signal tranducer and activator of transcription 1 (STAT1), E2F transcription factor-1 (E2F1), fibronectin-1 (FN1), interleukin-4 (IL-4), matrix metalloprotease-9 (MMP-9) and WAP four disulfide domain protein (WFDC-1) by QRTPCR. Additionally we performed immuno blot analysis and zymography to verify some of these candidate genes at the protein level. Computational analysis of gene function identified an anti-proliferative and altered immune function cellular phenotype in severe preeclamptic samples.
We have characterized the genome-wide mRNA expression changes associated with preeclampsia-specific genes in circulating maternal blood cells at the time of delivery. In addition to providing information relating to the biological basis of the preeclampsia phenotype, our data provide a number of potential biomarkers for use in the further characterization of this disease.
子痫前期是一种主要的产科并发症，影响母婴健康。虽然很明显，胎盘对子痫前期的发病机制有很大的贡献，但母体的贡献还不太清楚。因此，我们进行了全基因组转录组分析，以探讨外周血单个核细胞（PBMCs）中与疾病相关的母体基因表达模式的变化。
子痫前期的定义为妊娠高血压、蛋白尿和高尿酸血症。从无并发症妊娠的妇女（n=5）和子痫前期妊娠的妇女（n=5）中分离 PBMCs 的总 RNA。使用 Agilent 寡核苷酸微阵列进行基因表达分析。使用 Ingenuity Pathway Analysis 软件进行生物途径分析。定量实时 PCR（QRT-PCR）用于验证正常和子痫前期患者（n=12 各）中选定基因的基因表达变化。
我们总共鉴定了 368 个在子痫前期妇女中与正常对照相比差异表达的基因，假发现率（FDR）控制在 10%。在后续实验中，我们进一步分析了微阵列数据中鉴定为改变的一些基因的表达水平，包括存活素（BIRC5）、窖蛋白（CAV1）、GATA 结合蛋白-1（GATA1）、信号转导和转录激活因子 1（STAT1）、E2F 转录因子-1（E2F1）、纤维连接蛋白-1（FN1）、白细胞介素-4（IL-4）、基质金属蛋白酶-9（MMP-9）和 WAP 四硫域蛋白（WFDC-1）通过 QRT-PCR。此外，我们进行了免疫印迹分析和酶谱分析，以验证一些候选基因的蛋白水平。基因功能的计算分析确定了严重子痫前期样本中具有抗增殖和改变的免疫功能的细胞表型。
我们已经描述了与分娩时循环母体血细胞中特定于子痫前期的基因相关的全基因组 mRNA 表达变化。除了提供与子痫前期表型的生物学基础相关的信息外，我们的数据还提供了一些潜在的生物标志物，用于进一步表征这种疾病。
