ABSTRACT: Global transcriptional profiling confirmed failed decidualization of human Endometrial Stromal Cells (hESCs) obtained from non-pregnant women with a previous pregnancy complicated by severe preeclampsia (sPE).
Project description:In preeclampsia (PE), cytotrophoblast (CTB) invasion of the uterus and spiral arteries is often shallow. Thus, the placenta’s role has been a focus. We hypothesized that decidual defects are an important determinant of the placental phenotype. We isolated (human) endometrial stromal cells (hESCs) from non-pregnant donors with a prior pregnancy that was complicated by severe PE (sPE). Versus controls, they failed to decidualize as demonstrated by morphological criteria and the analysis of stage-specific antigens. These results were bolstered by showing that they were transcriptionally inert. Additionally, we used laser microdissection to isolate the decidua from tissue sections of the maternal-fetal interface. Transcriptional profiling revealed sPE-associated defects in gene expression. Also, decidual cells from sPE patients, which de-differentiated in vitro, failed to re-decidualize in culture. Immediately following isolation they released factors that inhibited CTB invasion, linking a possible cause to a known effect. These data suggested that failed decidualization is an important contributor to down regulated CTB invasion in sPE. Diagnosis of this defect prior to pregnancy would enable therapies that are designed to improve decidualization, a novel strategy for prevention.
Project description:The data presented here suggested that sPE is associated with impaired decidualization. Initial experiments involved an in vitro approach. hESCs were isolated from patients who developed sPE in a previous pregnancy or control women who had normal obstetric outcomes. Function was tested by comparing the ability of hESCs in the two groups to decidualize in culture. By morphological criteria and marker expression (PRL and IGFBP1), the cells in the formerly sPE group were decidualization resistant, which was confirmed by microarray analyses showing that they were transcriptionally inert.
Project description:Preeclampsia is a common complication of pregnancy that affects 4-5% of pregnant women around the world. At present, there is a lack of early identification of high-risk patients of preeclampsia in clinical practice, which restricts the development of disease prevention and treatment. Previous studies have indicated that plasma exosomal miRNAs in pregnant women could serve as biomarkers of preeclampsia, but few is focused on exosomal miRNAs from preeclampsia pregnancy with severe features(sPE). Therefore, we detected and compared the plasma exosomal miRNA profiles between normal pregancy and sPE to explore potential biomarkers and pathogenic mechanisms of sPE.
Project description:The aim of the study was to compare gene expression profiles in decidua basalis from cases with complicated pregnancies to those from healthy pregnant controls. Cases included pregnant women with preeclampsia (PE) and/or fetal growth restriction (FGR). Decidual tissue was obtained by vacuum suction of the placental bed after the placenta was delivered. Women with PE and/or FGR were included as cases. PE was defined as persistent hypertension (blood pressure (BP) ¡Ý 140/90 mm Hg), plus proteinuria (¡Ý0.3g/24 h or ¡Ý2+ according to a dipstick test), developing after 20 weeks of pregnancy. FGR implied birth weight under 2 standard deviations (SD) below the expected birth weight, as related to gestational age (GA) and sex. Due to tissue sampling procedures, only cases delivered by cesarean section (CS) were included. Healthy women with normal pregnancies, undergoing CS for various reasons considered irrelevant to the aim of this study (e. g. breech presentation and previous CS), served as controls. Tissue was immediately submerged in a RNA stabilisation solution (RNAlater, Ambion, Huntingdon, U.K.), incubated at 4¡ãC overnight and stored at -80¡ãC
Project description:The pathogenesis of preeclampsia superimposed on chronic hypertension is poorly understood relative to preeclampsia occurring in pregnant people without chronic hypertension. Placental transcriptomes in pregnancies complicated by de novo preeclampsia and superimposed preeclampsia have not been previously compared.
Project description:Background: Platelets may be pivotal mediators of the thrombo-haemorrhagic complications of preeclampsia (PE), linking inflammation and thrombosis with endothelial and vascular dysfunction. While gestational hypertension (GH) falls within the spectrum of hypertensive complications of pregnancy and is a risk factor for preeclampsia, it is unclear what biomarkers distinguish PE from GH. Aim: To identify specific plasma and platelet thrombo-inflammatory biomarkers indicative of preeclampsia and distinguish PE from GH. Methods: We performed multiplex immunoassays, assessed platelet and plasma proteomics and metabolomics data of PE patients, and compared with non-pregnant (NP), healthy pregnant (PC) and GH participants. Results: We report plasma proteins upregulated, enriched plasma metabolites and proteins distinctly overexpressed in platelets of PE and GH compared to NP and PC. Whilst procoagulation in PC may be fibrinogen driven, Inter-Alpha-Trypsin Inhibitors ITIH2 and ITIH3 were enriched in hypertensive complications of pregnancy (PE and GH), and fibronectin and S100A8/9 may be major procoagulant agonists in PE but not GH. In addition, platelet leucine-rich repeat-containing protein 27 and 42 (LRRC27/42) subunits of volume-regulated VRAC anion channels were markedly overexpressed in preeclampsia and may contribute to the heightened glucose sensitivity and the pro-thrombotic tendency of this disorder. Additionally, our multiplex immunoassays confirmed previous reports of increases in preeclampsia plasma cytokines, including SDF-1α, which can directly activate platelets; but also, i-309 and CTACK cytokines, whose effects on platelets we explored using STRING analysis. Conclusion: We identified biomarkers that may be monitored for preeclampsia onset and progression, and distinguish PE from GH. Also, through protein-protein interactions analysis, we generated a new hypothesis for platelets’ contribution to the thrombo-inflammatory states of preeclampsia.
Project description:Analysis of differences in gene expression in the mouse uterus during decidualization in the presence and absence of a conceptus. A deciduomal model previously shown to better mimick natural decidualization was used. The results reveal some, but very few, genes that are differentially expressed, which is unlike previous reports using other deciduomal models shown previously to not mimick natural decidualization. Total RNA obtained from isolated uterine segment tissue undergoing decidualization from pregnant or pseudopregnant mice. Three replicates per condition.
Project description:Maternal serum levels of calcyclin and heat shock protein 90 were compared throughout pregnancy from the first trimester till term among women with preeclampsia (PE) and age-matched normotensive pregnant controls (C). Serum samples from two different studies, a nested case-control study embedded in the Rotterdam periconception cohort and the Lepra Study both conducted at the Erasmus MC in Rotterdam. They were collected in the first, second and third trimester of pregnancy in 43 patients with preeclampsia, consisting of 20 early-onset and 23 late-onset preeclampsia, and 46 normotensive pregnant controls. A serum based 2D LC-MS assay on Parallel Reaction Monitoring mode using a high resolution tribrid mass spectrometer was used to quantify both calcyclin and heat shock protein 90.
Project description:Normal pregnancy requires adaptations of the maternal vasculature. During preeclampsia these adjustments are not well established, resulting in maternal hypertension and proteinuria. The effects of preeclampsia on the maternal vasculature are not yet fully understood. We aimed to identify gene expression differences in the aorta between non pregnant, healthy pregnant, and experimental preeclamptic rats using a genome wide approach. Whole aortic tissue was isolated from rats with low-dose LPS-induced preeclampsia, healthy pregnant and non-pregnant rats. Gene expression was measured by a whole genome microarray.
Project description:Decidualization is critical for the embryonic implantation and successful pregnancy. ATRA can suppress in-vitro decidualization of human endometrial stromal cells (hESCs) induced by MPA and estrogen treatment. However, the mechanism by which RA suppressed estrogen and progesterone induced decidualization of mESCs is not clear. We used microarrays to investigate the mechanism by which all-trans RA (ATRA) regulates the decidualization of endometrial stroma cells (mESCs). mESCs were isolated at day 4 of pseudopregnancy and cultured with administration of E2 and P4 in the presence or absence of ATRA for 72h.
