Etiology Of Preeclampsia Research Articles

Role of UMOD Promoter Polymorphism in the Etiology of Preeclampsia

To evaluate the association between preeclampsia (PE) and the single nucleotide polymorphism (SNP) rs13333226, located in the promoter region of the UMOD gene. A total of 1248 pregnant Han Chinese women (716 controls and 532 patients with PE) were included in this study. Genotyping of the rs13333226 polymorphism was performed by real-time PCR using a TaqMan-minor groove binder (MGB) probe assay. No significant differences were detected in the allele (p = 0.62, OR = 1.08, 95% CI = 0.81-1.44) and genotype frequencies of rs13333226 (padditive = 0.38, pdominant = 0.45, precessive = 0.31) between cases and controls. When patients were divided into subgroups, no association was found with mild preeclampsia (M PE), severe preeclampsia (S PE), early onset PE, or late-onset PE. Furthermore, no significant differences were detected in the genotype and allele frequencies of rs1333226 between patients with M PE and S PE (p > 0.05) or between patients with late and early onset of the disease (p > 0.05). UMOD rs13333226 does not appear to be associated with PE in Han Chinese women.

Unsupervised Placental Gene Expression Profiling Identifies Clinically Relevant Subclasses of Human Preeclampsia.

Preeclampsia (PE) is a complex, hypertensive disorder of pregnancy, demonstrating considerable variability in maternal symptoms and fetal outcomes. Unfortunately, prior research has not accounted for this variability, resulting in a lack of robust biomarkers and effective treatments for PE. Here, we created a large (N=330) clinically relevant human placental microarray data set, consisting of 7 previously published studies and 157 highly annotated new samples from a single BioBank. Applying unsupervised clustering to this combined data set identified 3 clinically significant probable etiologies of PE: "maternal", with healthy placentas and term deliveries; "canonical", exhibiting expected clinical, ontological, and histopathologic features of PE; and "immunologic" with severe fetal growth restriction and evidence of maternal antifetal rejection. Moreover, these groups could be distinguished using a small quantitative polymerase chain reaction panel and demonstrated varying influence of maternal factors on PE development. An additional subclass of PE placentas was also revealed to form because of chromosomal abnormalities in these samples, supported by array-based comparative genomic hybridization analysis. Overall, our findings represent a new paradigm in our understanding of the origins and maternal-placental contributions to the pathology of PE. The study of PE represents a unique opportunity to access human tissue associated with a complex hypertensive disorder, and our novel approach could be applied to other hypertensive and heterogeneous human diseases.

Research of MFG-E8, OPG and SOCS-3 on preeclampsia

Abstract Preeclampsia is a specific disease and closely associated with systemic inflammatory response. The aim of this study is research to possible role of some inflammatory factors and the controlling factor in inflammation in the etiology of preeclampsia or reflection of this situation on placental tissue. Three parameters were included in the study regarded as potential risk factors: Milk Fat Globul Epidermal Growth Factor-8 (MFG-E8); is an endometrial epithelial proteins and regulates inflammation and apoptosis. MFG-E8 is an important in the development of endometrium and chorionic villus. Osteoprotegerin (OPG); is pro-angiogenic factor, it can be important for plasental angiogenesis. Suppressor of cytokine signaling-3 Receptor (SOCS-3) has a role on prevents the inflammation. Serum and placental tissue samples were used as materials. Control group is composed of 50 healthy women volunteers with no history of preeclampsia and 28–34th weeks of pregnancy. The experimental group consisted 40 preeclamptic women in 28–34th week of pregnancy. All of the measurement analyzed through the ELISA (Enzyme-Linked Immuno Sorbent Analysis) method and findings evaluated comparatively. MFG-E8, SOCS-3 and OPG levels analysed with ELISA in the serum and placental tissue. OPG levels in serum raised and displayed statistically significant difference with respect to control group (p<0.01). In respect to the serum MFG-E8 and SOCS-3 are not observed any difference between two groups. MFG-E8 tissue levels are statistically significantly low in preeclamptic group with respect to healthy control group and that there is no changes in the levels of SOCS-3 (p<0.01).

Expression and clinical significance of calcitonin receptor stimulating peptide-3 (CRSP-3) in placenta tissue of preeclampsia patients

Objective To study clinical significance and the difference of calcitonin receptor stimulating peptide-3 (CRSP-3) expression in placenta tissue between normal pregnancies and patients with preeclampsia. Methods CRSP-3 expression position in placenta tissues of 50 cases with preeclampsia (preeclampsia group: 25 cases of mild preeclampsia and 25 cases of severe preeclampsia) and 30 cases of normal late pregnancy (control group) was detected using immunohistochemical methods. CRSP-3 mRNA and protein expression in placenta tissues was detected using Real-time PCR and Western blotting method. Results Syncytiotrophoblast and cytotrophoblast cells expressed CRSP-3 in placenta tissue of preeclampsia and normal pregnancy patients. The expression levels of CRSP-3 mRNA and protein in placenta tissue of preeclampsia patients were higher than those in control group (P 0.05). Conclusion The up-regulation of CRSP-3 in placenta of preeclamptic patients may be associated with the etiology of preeclampsia, but may have no relation with progress and degree of preeclampsia.

Leukemia inhibitory factor promote trophoblast invasion via urokinase-type plasminogen activator receptor in preeclampsia.

Preeclampsia is a pregnancy-related syndrome which can cause perinatal mortality and morbidity. Inadequate invasion by trophoblast cells may lead to poor perfusion of the placenta, even result in preeclampsia. Understanding the molecular mechanisms underlying placentation facilitates the better intervention of preeclampsia. Urokinase-type plasminogen activator receptor (uPAR) is involved in the physiological and pathological processes. Leukemia inhibitory factor (LIF) is an important regulator in the establishment of pregnancy. However, the expression of uPAR in preeclamptic patients and its relationship with LIF remains unclear. In the current study, we found that the level of uPAR was relatively lower in the placentas from preeclamptic patients as compared with normal pregnant women. LIF promoted trophoblast cell outgrowth by upregulating uPAR in an explants culture, and LIF also enhanced migration and invasion potential through uPAR in trophoblast JAR and JEG-3 cell lines, and with increased gelatinolytic activities of matrix metalloproteinase 2 (MMP-2). The effect of LIF and uPAR on trophoblast migration and invasion was mediated by PI3K/AKT signaling pathway. Our data indicates the roles of LIF in promoting trophoblast migration and invasion through uPAR and suggest that abnormal expression of uPAR might be associated with the etiology of preeclampsia.

Abstract P089: Pre-eclampsia and Risk of Subsequent Hypertension: In an American Indian Population

Background and Objectives: Preeclampsia (PE) is a pregnancy-specific disorder that affects 2-8% of all pregnancies and as many as 60,000 annual maternal mortalities globally. A number of proposed pathophysiologic mechanisms relate to similar factors implicated in cardiovascular disease, such as endothelial dysfunction, inflammation, insulin resistance and impaired renal regulation. Pre-eclampsia has been associated with subsequent hypertension, cardiovascular disease and related mortality in later life. Methods: The 3 most recent blood pressures, body mass index (BMI) and use of hypertensive medications were recorded from electronic medical records of routine clinic visits for 130 cases and 289 controls previously identified and carefully characterized for a study of the etiology and complications of PE. Comparison between case and control groups was made using the student t test for continuous variables and chi-square testing for discrete variables. Linear regression analysis was used to evaluate the effects of risk factors on subsequent systolic and diastolic blood pressure (SBP and DBP respectively). Results: Follow-up measurements occurred a mean of 13.11 years (range 3.6 to 36.7 years) post PE pregnancy. Cases were of similar age (35.9 vs 36.1 years, p=0.82); but with significantly higher BMI (34.1 vs 31.6, p<0.002), systolic (127.2 vs 121.3 mm Hg, p<0.001 and diastolic (76.2 vs 72.6 mm Hg, p<0.001) blood pressure, and more likely to be using hypertensive medications (35/130, 27% vs 25/289, 9%, p<0.001). Linear regression showed significant and independent association between BMI (SBP β=0.47, DBP β=0.29), age (SBP β=0.19, DBP β=0.14), previous history of PE (SBP β=4.47, DBP β=2.71) and current systolic and diastolic blood pressure (all p values <0.002). Analysis of the quartile with follow-up of less than 7.19 years also shows independent association of prior PE with both SBP (β=6.54, p<0.006) and DBP (β=4.96, p<0.001). Discussion and Conclusions: PE appears to confer risk of subsequent hypertension on American Indian women after a period of at least 13 years. This risk is independent of additional risk factors such as increased BMI and age. The risk is apparent soon after the affected pregnancy and there are undoubtedly cases where the hypertension of PE fails to resolve post partum.

Spectrum of Factors Triggering Endothelial Dysfunction in PIH.

Pre-eclampsia (PE) is a major cause of maternal and fetal/neonatal mortality and morbidity. The aetiology and pathogenesis of PE is yet to be completely understood. Evidence shows that, Endothelial Dysfunction (ED) plays a pivotal role in the genesis of this multi-system disorder that develops in PE and eclampsia. To determine the circulating levels of factors Malondialdehyde (MDA), Ferric Reducing Ability of Plasma-α (FRAP), Tumour Necrosis Factor (TNF-α), sFlt-1, VEGF, PlGF, Nitric Oxide (NO) that influence the ED. Study groups consisted of Normotensive pregnant women (N), preeclamptic women (PE) and eclamptic women (E) with 100 subjects in each group in the 3rd trimester of pregnancy. They were investigated for MDA, FRAP, TNF-α, sFlt-1, VEGF, PlGF, NO. Statistical analysis was done using Analysis of Variance (ANOVA). When compared to controls MDA, TNF-α, sFlt-1 levels were found to be significantly high and FRAP, VEGF, PIGF and NO levels were significantly low in PE and E group. E showed a significantly high level of MDA, TNF-α, sFlt-1 and low levels of FRAP, VEGF, PIGF, NO when compared to PE group. Our study substantiated the fact, that oxidative stress, imbalance between anti-angiogenic factors and pro- angiogenic factors exists in Pregnancy Induced Hypertension (PIH) condition. This imbalance is directly related to the ED, the hallmark of PE. So oxidative stress, VEGF, PlGF and sFlt-1 can be used as markers to analyze the onset and progression of the disease.

11β-Hydroxysteroid Dehydrogenase 2 in Preeclampsia.

Preeclampsia is a serious medical problem affecting the mother and her child and influences their health not only during the pregnancy, but also many years after. Although preeclampsia is a subject of many research projects, the etiology of the condition remains unclear. One of the hypotheses related to the etiology of preeclampsia is the deficiency in placental 11β-hydroxysteroid dehydrogenase 2 (11β-HSD2), the enzyme which in normal pregnancy protects the fetus from the excess of maternal cortisol. The reduced activity of the enzyme was observed in placentas from pregnancies complicated with preeclampsia. That suggests the overexposure of the developing child to maternal cortisol, which in high levels exerts proapoptotic effects and reduces fetal growth. The fetal growth restriction due to the diminished placental 11β-HSD2 function may be supported by the fact that preeclampsia is often accompanied with fetal hypotrophy. The causes of the reduced function of 11β-HSD2 in placental tissue are still discussed. This paper summarizes the phenomena that may affect the activity of the enzyme at various steps on the way from the gene to the protein.

Serum copper and zinc levels in preeclamptic Nigerian women.

Background:The exact etiology of preeclampsia remains unknown even though several studies have been done. Some studies have shown that supplementation of zinc (Zn) and copper could ameliorate the effects of preeclampsia while other studies did not establish the beneficial role of these elements in preeclampsia. The objective of this study was to compare the Zn and copper levels in the serum of Nigerian women with or without preeclampsia.Materials and Methods:In this study, serum Zn and copper levels were determined using atomic absorption spectrometry in 54 patients and 48 healthy normotensive pregnant women. The mean, standard deviation, Student's t-test, and Pearson correlation were employed.Results:Serum Zn was significantly lower in patients than controls (8.27 ± 0.60 vs. 12.16 ± 1.83 μmol/l. P <0.001) (t-test). Serum copper was also significantly lower in patients than controls (8.14 ± 1.80 vs. 16.62 ± 3.17 μmol/l, P < 0.001).Conclusion:There was a significant reduction in the levels of Zn and copper in patients with preeclampsia. Dietary supplementation of these trace elements may help to prevent preeclampsia.

Inventory of Novel Animal Models Addressing Etiology of Preeclampsia in the Development of New Therapeutic/Intervention Opportunities.

Preeclampsia is a pregnancy-related disease afflicting 3-7% of pregnancies worldwide and leads to maternal and infant morbidity and mortality. The disease is of placental origin and is commonly described as a disease of two stages. A variety of preeclampsia animal models have been proposed, but all of them have limitations in fully recapitulating the human disease. Based on the research question at hand, different or multiple models might be suitable. Multiple animal models in combination with in vitro or ex vivo studies on human placenta together offer a synergistic platform to further our understanding of the etiology of preeclampsia and potential therapeutic interventions. The described animal models of preeclampsia divide into four categories (i) spontaneous, (ii) surgically induced, (iii) pharmacologically/substance induced, and (iv) transgenic. This review aims at providing an inventory of novel models addressing etiology of the disease and or therapeutic/intervention opportunities.

Analysis of the gene polymorphism of aldosterone synthase (CYP11B2) and atrial natriuretic peptide (ANP) in women with preeclampsia

ObjectivePreeclampsia (PE) is a major cause of mortality of mothers, fetuses and newborns around the world. The etiology of preeclampsia has not yet been clarified, but many studies indicate a multifactorial basis of PE. Aldosterone synthase (CYP11B2) is responsible for synthesis of aldosterone responsible for regulating blood pressure. Similarly, natriuretic peptide (ANP) regulates blood pressure through a variety of mechanisms affecting the sodium concentration and the amount of extracellular fluid. Currently, attention is paid to the role of the polymorphisms in the expression level of these genes. The aim of the study was to determine the frequencies of genotypes and alleles for polymorphisms of −344C>T CYP11B2 gene and 2238T>C ANP gene in women with preeclampsia and healthy pregnant women from the Caucasian population. Study designThe study included a group of 165 pregnant women (59 women with preeclampsia and 109 healthy pregnant women). DNA was extracted from peripheral blood. Determination of the polymorphism of −344C>T CYP11B2 gene and 2238T>C ANP gene was performed by PCR-RFLP method. ResultsThe results showed that the frequencies of the TC and CC genotypes of 2238T>C polymorphism in ANP gene were significantly higher in patients with PE compared to control group. For −344C>T polymorphism of CYP11B2 gene, the frequency of TT genotype was significantly higher in patients with hypertension than in controls (32.2% vs. 23.58%). ConclusionsOur findings showed that gene polymorphism of CYP11B2 (−344C>T) and ANP (2238T>C) may be associated with developing PE during pregnancy.

Abstract P612: TLR3-Induced Placental miR-155 Contributes to Preeclampsia by Augmenting Inflammation

Hypertensive disorders of pregnancy including preeclampsia (PE) affect ~15% of all pregnancies. We have previously demonstrated that excessive maternal immune system activation via Toll-like receptor 3 (TLR3) contributes to the development of PE-like symptoms such as hypertension, endothelial dysfunction, and proteinuria in mice only when pregnant. Recently, increased miR-155 (an inflammation-related miRNA) expression was reported in placentas from women with PE compared to normotensive women suggesting a pathophysiological role of miR-155 in PE. Whether miRNAs play a role in the etiology of PE by augmenting inflammation is unknown. We confirmed miR-155 expression was up-regulated in placentas of poly I:C (a TLR3 agonist) treated wild type mice by microarray (P-PIC: 1.56 fold, p&lt;0.5 vs controls) and qRT-PCR analyses (P-PIC: 3.15 fold, p&lt;0.05 vs. controls). Based on our data we hypothesized that TLR3 activation induces placental miR-155 expression that augments maternal inflammation leading to PE. Transcription factors NF-κB and AP-1 which are known to bind to the promoter of miR-155 were increased in placentas of P-PIC mice. TargetScan, a target identification algorithm predicted suppressor of cytokine signaling 1 (SOCS1) as a putative target of miR-155. SOCS1 prevents overactivation of immune responses but failed to increase in P-PIC mice. ICAM, a marker of inflammation increased significantly in P-PIC mice. P-PIC TLR3 KO mice did not develop hypertension and proteinuria as well as did not exhibit increased placental levels of NF-κB, AP-1, miR-155 expression and ICAM or decreased levels of SOCS1. To determine the placental etiology, human cytotrphoblasts (CTBs) were treated with poly I:C and NF-κB, AP-1 transcription factor levels and miR-155 expression were increased significantly while SOCS1 levels decreased. Together, these data suggest that TLR3 activation increases placental miR-155 expression which downregulates SOCS1 levels and increases inflammation which may contribute to the development of PE.

Abstract P613: The Effect of Metabolic Factors on Hypoxia-Induced sFlt-1 Secretion in Rat Placental Villi

Although the etiology of preeclampsia (PE) remains unclear, evidence indicates that impaired trophoblast invasion followed by placental ischemia/hypoxia promotes the release of placental anti-angiogenic factors, such as soluble fms-like tyrosine kinase-1 (sFlt-1), into the maternal circulation. sFlt-1 blocks the pro-angiogenic actions of vascular endothelial growth factor to elicit maternal endothelial dysfunction and ultimately hypertension. Obesity is a major risk factor for PE. In addition, increased circulating metabolic factors, such as leptin and insulin have been associated with PE. However, the mechanisms whereby obesity and its related metabolic factors increase the risk for the development of PE are unknown. The aim of this study was to evaluate whether chronic leptin or insulin exposure exacerbate hypoxia-induced sFlt-1 secretion from rat placental villi. In order to address this question, placental villous explants were isolated from placentas of normal pregnant rats (n=4, 3 placentas per rat) and pregnant rats treated with either leptin (0.5 mg/kg/min i.p.; n=3, 3 placentas per rat) or insulin (1.5 mU/kg/min s.c.) supplemented with 20% glucose in drinking water (n=3, 3 placentas per rat) from gestational day 14 to 19. Placental explants were then incubated for 48 h at 37 °C under normoxia (6% O2) or hypoxia (1% O2) and sFlt-1 secretion in cultured media was measured by ELISA. While hypoxia significantly enhanced sFlt-1 release of explants from normal pregnant rats compared with normoxia (3224±224 vs 4251±236 pg/mg; P&lt;0.05), explants from chronic hyperleptinemic (3197±178 vs. 3762±317 pg/mg) or euglycemic hyperinsulinemic (4066±186 vs. 4251±213 pg/mg) pregnant rats secreted similar sFlt-1 levels under normoxic and hypoxic conditions, respectively. Additionally, chronic leptin or insulin treatments did not exacerbate the effect of hypoxia on sFlt-1 release. In conclusion, our in vitro studies with placental villi from chronic hyperleptinemic or euglycemic hyperinsulinemic pregnant rats showed no exacerbation of hypoxia-induced sFlt-1 secretion.

Abstract 129: A Biopolymer-Stabilized VEGF Chimera Reverses Angiogenic Imbalance in vitro in the Reduced Uterine Perfusion Pressure Model of Preeclampsia

Preeclampsia (PE) is a hypertensive disorder that complicates approximately 5-8% of all pregnancies in the U.S. The unknown etiology of PE develops from molecular dysfunction at the maternal-placental interface, where inflammatory, oxidative stress, and anti-angiogenic pathways are initiated. The resulting hypoxic in utero environment contributes to placental ischemia from which the anti-angiogenic factor soluble fms-like tyrosine kinase-1 (sFlt-1) is released. A truncated isoform, sFlt-1 consists solely of the extracellular domain of the vascular endothelial growth factor (VEGF) receptor 1, and hence lacks both catalytic and regulatory activities. However, sFlt-1 retains a high affinity for VEGF and sequesters this ligand from binding with fully functional VEGF receptors, thus prevents the activation of angiogenic remodeling pathways. The rodent model of PE, reduced uterine perfusion pressure (RUPP), exhibits the sFlt-1 pathophysiology observed in human PE. In an effort to counteract excessive sFlt-1 production and restore angiogenic balance, we have constructed a VEGF chimera fused to an Elastin-like Polypeptide (ELP) carrier with increased in vivo half-life and stability which retains full VEGF signaling activity. When human umbilical vein vascular endothelial cells (HUVECs) are exposed to serum from normal pregnant or RUPP-treated rats, tube formation on extracellular matrix is inhibited 31% (± 2%) by the RUPP serum. This inhibition is reversed when ELP-VEGF, but not ELP control, is added to the culture medium ( p = 0.0007, one-way ANOVA with Bonferroni multiple comparison), suggesting that ELP-VEGF counteracts the anti-angiogenic factors present in RUPP serum. We also characterized the pharmacokinetics, biodistribution, and placental transfer of ELP-VEGF in the pregnant Sprague Dawley rat. These studies indicate that ELP-delivered VEGF has potential for counteracting the circulating anti-angiogenic factors in maternal plasma.

Upregulation of the Tim-3/Gal-9 pathway and correlation with the development of preeclampsia

ObjectiveIt is well documented that an imbalance in immune tolerance at the maternal–fetal interface is likely to play an essential role in the etiology of preeclampsia. However, the mechanisms underlying immune tolerance during preeclampsia are still poorly understood. Tim-3, a Th1-specific cell surface molecule, is a relatively newly described molecule with important immunological functions. It can regulate Th1 responses with its ligand galectin-9 (Gal-9), and has become an attractive candidate for exploring the pathogenesis of preeclampsia. Study designTwenty normal pregnancies and 20 preeclamptic pregnancies were enrolled in the present study. We examined the expression and function of Tim-3/Gal-9 in decidual tissue at the RNA and protein levels. In order to analyze their correlation with the development of preeclampsia, we measured the expression of Tim-3 on peripheral blood leukocytes using flow cytometry. IFN-γ, IL-10, and IL-17 in the peripheral blood plasma were measured by ELISA. ResultsTim-3/Gal-9 was upregulated in decidual tissue of preeclamptic vs. normotensive pregnancies. There was a significantly increased Th1 and Th17 response in PE as demonstrated by the upregulated levels of IFN-γ/IL-17, whereas IL-10 secreted by Th2 cells was sharply reduced. ConclusionsThe present study showed that an abnormal Tim-3/Gal-9 pathway was able to facilitate the development of preeclampsia. Our data uncovered a novel mechanism by which the Tim-3/Gal-9 pathway regulates immune responses, and we now identify this pathway as a potential therapeutic target in preeclampsia.

Genome-Wide Mapping of 5mC and 5hmC Identified Differentially Modified Genomic Regions in Late-Onset Severe Preeclampsia: A Pilot Study.

Preeclampsia (PE) is a leading cause of perinatal morbidity and mortality. However, as a common form of PE, the etiology of late-onset PE is elusive. We analyzed 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) levels in the placentas of late-onset severe PE patients (n = 4) and normal controls (n = 4) using a (hydroxy)methylated DNA immunoprecipitation approach combined with deep sequencing ([h]MeDIP-seq), and the results were verified by (h)MeDIP-qPCR. The most significant differentially methylated regions (DMRs) were verified by MassARRAY EppiTYPER in an enlarged sample size (n = 20). Bioinformatics analysis identified 714 peaks of 5mC that were associated with 403 genes and 119 peaks of 5hmC that were associated with 61 genes, thus showing significant differences between the PE patients and the controls (>2-fold, p<0.05). Further, only one gene, PTPRN2, had both 5mC and 5hmC changes in patients. The ErbB signaling pathway was enriched in those 403 genes that had significantly different5mC level between the groups. This genome-wide mapping of 5mC and 5hmC in late-onset severe PE and normal controls demonstrates that both 5mC and 5hmC play epigenetic roles in the regulation of the disease, but work independently. We reveal the genome-wide mapping of DNA methylation and DNA hydroxymethylation in late-onset PE placentas for the first time, and the identified ErbB signaling pathway and the gene PTPRN2 may be relevant to the epigenetic pathogenesis of late-onset PE.

P40. MIR-21 and mir-221 overexpression in placental tissue of preeclamptic patients

Introduction Preeclampsia is a hypertensive disorder potentially leading to severe complications in mother and the fetus. Etiology of preeclampsia is multifactorial including partially genetic and immunological pathways; the process of apoptosis is also involved in the development of preeclampsia. MicroRNAs (miRNAs) - small noncoding molecules- are emerging as critical regulators of biological function (including apoptosis), and alterations to the miRNAome have been described in the context of pregnancy and PE. Objectives Goal of this work was to assess expression of pre-selected miRNAs in preeclamptic placental samples and comparison with samples from physiological pregnancies. We designed a 96-well reaction plate to analyze 21 miRNAs in duplicates described to be involved in the apoptosis regulation and expressed in placenta: pro- apoptotic miR-1, let-7c, let-7g, mir-200c, mir-143, mir-205, mir-122, mir-409-3p, mir-449, mir-708, mir-149, mir-204,mir-133 and anti-apoptotic : mir-214, mir221, mir 222, and miRNAs with both the anti-apoptotic and apoptotic targets mir29a and mir29c. Methods The normalization was performed against RNU44. The miRNAs were extracted from placental tissue obtained after delivery from both preeclamptic women and healthy controls. After stem-loop primer reverse transcription in one tube, the samples were analyzed on the plates and evaluated by delta Ct algorithm. The aberrantly expressed miRNAs were subsequently validated by TaqMan MicroRNA Assay by relative quantification with the standard curves. Results Overexpression of miRNA was observed in mir-122, mir-21, mir-221, mir-29a, mir-29c and mir-449. The validation was performed on 13 preeclamptic placental samples and 6 placental samples from physiological pregnancies; significant overexpression of mir-21 and mir-221 with p =0.0055 (CI 95% 25.2–123.47) and p =0.0047 (CI 95% 12.16–57.20) was observed, respectively. Conclusion Analysis of miRNA deregulation can help in the identification of deregulated signaling in preeclamptic placentas for detection of new, potentially usable biomarkers. Even though authors would like to point out the necessity of validation on a larger sample cohort.

STA54. Preeclampsia – Myths are still stronger than scientific data

Introduction During pregnancy close interactions between the maternal and fetal system exist with the placenta as the organ situated between the two individuals. In cases where this crosstalk is disturbed, preeclampsia may develop. During the last decades a number of hypotheses have been developed to describe the etiology of this syndrome. However, so far all of them have failed. Objectives One of the current hypotheses – although wrong as well – is still strongly supported by the scientific community although it has become clear in recent years that a variety of arguments that are needed to support this hypothesis are not valid at all. Methods This list comprises the following arguments, known to be wrong today: (1) Failure in trophoblast invasion is a prerequisite for preeclampsia. (2) Placental hypoxia is present in cases suffering from preeclampsia. (3) During preeclampsia, the placenta is the main source for angiogenic factors in maternal blood. Additionally, another important issue needs attention: (4) The quality of a biomarker only depends on its concentration in maternal blood. Results These four arguments and issues will be discussed. It will become clear that (1) changes in trophoblast invasion are not related to preeclampsia; (2) placental hypoxia does not exist at all in cases with preeclampsia; (3) the maternal vascular system is the main source for angiogenic factors; and (4) pre-analytics are increasingly important to maintain the quality of samples to test for biomarkers. Conclusion Due to the availability of new data it is more than needed to update the hypotheses on the etiology of preeclampsia. It is quite detrimental to keep with old but no longer valid hypotheses and hindering the rise of newer and better hypotheses. Therefore, the above mentioned arguments including trophoblast invasion, placental hypoxia and the source of angiogenic factors need to be taken into account when further discussing the etiology of preeclampsia.

P42. TNF-ALPHA gene polymorphism in pathology of preeclampsia

Introduction Preeclampsia (PE) is a life-threatening complication of pregnancy associated with a high rate of maternal and perinatal morbidity and/or mortality worldwide. If untreated, it can progress to eclampsia, which can result in the death of mother, fetus or both. Objectives Etiology of PE is still uncertain, however recently the role of immune system takes on importance. Thus the role of central cytokine, tumor necrosis factor (TNF) involved in inflammation processes become widely investigated in obstetric disorders. Following this the present study is to investigate the effect of TNF-alpha gene G308A (rs1800629) polymorphism on disease risk, renal functions, microvascular permeability, endothelial cell dysfunction and organ involvement in women with preeclampsia. Methods Initially a 102 3 rd trimester pregnant women (preeclamptic-cases and healthy controls) with singleton pregnancy were invited for participation of which 76 were genotyped for TNF-alpha G308A polymorphism, evaluated for plasma levels of sVCAM-1, fibronectin, TNF-alpha and correlated to profile of preeclampsia. The odds ratio (OR) and 95% confidence intervals obtained from unconditional logistic regression were used to set association between TNF-apolymorphism and PE risk. For continuous variables we applied Student's t -test and for categorical variables the Pearson x 2 or Fisher's exact test. The two-way ANOVA with Bonferroni correction was used in multivariate analysis. Results The A allele was more frequent in cases than controls (22.4% vs. 13.2%) what increased disease risk (OR=2.73). Maternal serum levels of TNF-alpha, sVCAM-1 and fibronectin were significantly increased in cases (855.8±385.1pg/mL, 1243±671ng/mL, 0.308±0.231g/L) compared to controls (301.1±156.1pg/mL, 651±250ng/mL, 0.218±0.101g/L, p p p =0.031; respectively) and levels shoved increasing trend with mutant allele in genotype. Moderate and severe proteinuria was higher in rs1800629 allele A subjects compared to G/G carriers (53.8% vs. 14.3%, p p Conclusion This study revealed possible association between clinical and laboratory manifestations of preeclampsia and TNF-alpha gene G308A (rs1800629) polymorphism.

22: A Prospective Assessment of Neurodevelopment in Children Following a Pregnancy Complicated by Severe Preeclampsia

Preeclampsia (PE) affects 2–7% of otherwise healthy women, and is diagnosed by newly elevated BP after 20 weeks gestation, with associated proteinuria. The etiology of PE is largely unknown. Many theories have been proposed, including mechanisms involving oxidative stress, angiogenic imbalance, and immunologic intolerance between feto-placental and maternal tissue. Delivery is the only definitive cure, with other treatments focused on symptom management. Gestational hypertension occurs in 6–17% of pregnancies, and is defined as newly elevated blood pressure (BP), with the absence of proteinuria, after 20 weeks gestation in a previously normotensive individual. These hypertensive disorders of pregnancy are associated with adverse health outcomes including perinatal deaths, preterm birth, intrauterine growth restriction (IUGR), neonatal morbidity, and infants that are small for gestational age. Previous studies have demonstrated a link between PE and neurodevelopmental delay; however there has been disagreement on whether this effect is protective or harmful. Additionally the independent effects of gestational age (GA), birth weight, and PE are unclear, and the vast majority of studies have been retrospective or cross-sectional in nature. To prospectively examine whether infants of women with a PE pregnancy, compared to infants of women without a PE pregnancy, have differences in anthropometric measurements and/or neurodevelopmental performance. Women were recruited into this longitudinal prospective cohort following a pregnancy with (n=129) or without PE (n=140). Anthropometric measurements were performed on offspring at birth, one, three, and five years of age. The Ages and Stages Questionnaire (ASQ) was completed yearly, until age five. Anthropometrics did not differ between severe PE and controls after birth. The proportion of ASQ categories failed was significantly different in years 1, 3, and 4. Comparison of the distribution of number of categories failed revealed that severe PE children failed significantly more categories at year 1. A subgroup analysis indicated that preterm birth significantly contributes to this relationship, which persists through year 5. Logistic regression indicated that severe PE and IUGR are associated with increased odds of failure, while increased GA is protective. Growth delays experienced by children born to mothers with PE are not found beyond one year of age; however neurodevelopmental changes persist with preterm birth.