Increased decidual mRNA expression levels of candidate maternal pre-eclampsia susceptibility genes are associated with clinical severity

Introduction

Pre-eclampsia (PE) has a familial association, with daughters of women who had PE during pregnancy having more than twice the risk of developing PE themselves. Through genome-wide linkage and genetic association studies in PE-affected families and large population samples, we previously identified the following as positional candidate maternal susceptibility genes for PE; ACVR1, INHA, INHBB, ERAP1, ERAP2, LNPEP, COL4A1 and COL4A2. The aims of this study were to determine mRNA expression levels of previously identified candidate maternal pre-eclampsia susceptibility genes from normotensive and severe PE (SPE) pregnancies and correlate mRNA expression levels with the clinical severity of SPE.

Methods

Third trimester decidual tissues were collected from both normotensive (n = 21) and SPE pregnancies (n = 24) and mRNA expression levels were determined by real-time PCR. Gene expression was then correlated with several parameters of clinical severity in SPE. Statistical significance was determined by Mann–Whitney U test and Spearman's Correlation.

Results

The data demonstrate significantly increased decidual mRNA expression levels of ACVR1, INHBB, ERAP1, ERAP2, LNPEP, COL4A1 and COL4A2 in SPE (p < 0.05). Increased mRNA expression levels of several genes – INHA, INHBB, COL4A1 and COL4A2 were correlated with earlier onset of PE and earlier delivery of the fetus (p < 0.05).

Conclusion

These results suggest altered expression of maternal susceptibility genes may play roles in PE development and the course of disease severity.     

Placental expression and serum levels of cytokeratin-18 are increased in women with preeclampsia

OBJECTIVE: To evaluate placental expression and serum cytokeratin-18 in women with preeclampsia. METHODS: Serum cytokeratin-18 was evaluated in 44 women with preeclampsia and 44 healthy pregnant women using an immunoradiometric assay. Placental expression of cytokeratin-18 was investigated in specimens from 23 women with preeclampsia and 20 healthy pregnant women by immunohistochemistry. RESULTS: Median serum cytokeratin-18 in women with preeclampsia and healthy pregnant women was 106.7 and 76.0 U/L, respectively (P =.02). Among women with preeclampsia, serum cytokeratin-18 was significantly associated with severity of disease (P =.001) and showed a sensitivity (standard error) and specificity (standard error) of 85% (7%) and 65% (12%), respectively. In placental specimens, the cytoplasm of the syncytiotrophoblast stained positive for cytokeratin-18 with strong and widespread staining in 83% and 45% of placental specimens of women with preeclampsia and healthy pregnant women, respectively (P =.01). CONCLUSION: Elevated serum cytokeratin-18 values are associated with disease severity in women with preeclampsia. Our data provide additional evidence that the placenta might be the source of the elevated serum cytokeratin-18 values in women with preeclampsia.     

First-trimester maternal serum metastin, placental growth factor and chitotriosidase levels in pre-eclampsia

Objective

To investigate whether the serum levels of metastin and PIGF and chitotriosidase activity early in pregnancy differ in women who develop pre-eclampsia from those who remain normotensive.

Study design

A retrospective case–control study of prospectively collected data. Thirty healthy pregnant women and 31 women with pre-eclampsia were included in the study. Serum samples were collected at 11–14 weeks and stored at −70 °C. Levels of metastin, PIGF and chitotriosidase activity were measured in serum from pregnant women with subsequent development of pre-eclampsia and matched controls.

Results

Mean maternal serum metastin (1554 ± 385 pmol/L vs 1995 ± 375 pmol/L, p < 0.001) and PIGF (111.9 ± 7.0 pg/mL vs 124.9 ± 13.5 pg/mL, p < 0.001) levels were significantly lower and chitotriosidase activity was significantly higher (681.6 ± 248.3 nmol/mL/h vs 527.7 ± 223.1 nmol/mL/h, p < 0.01) in women who subsequently developed pre-eclampsia than in those who remained normotensive. The areas under the curve equal to 0.797, 0.831 and 0.681 (p < 0.001, p < 0.001 and p < 0.01) for metastin, PIGF, and chitotriosidase respectively were determined for the prediction of pre-eclampsia.

Conclusions

Metastin and PIGF levels and chitotriosidase activity are altered in the first trimester serum of women destined to become pre-eclamptic, reflecting placental dysfunction. Metastin, like PIGF, may have a potential to be used as a first-trimester biomarker of pre-eclampsia.     

Umbilical cord serum activin A levels are increased in pre-eclampsia with impaired blood flow in the uteroplacental and fetal circulation

The aims of the present study were to evaluate the umbilical cord serum activin A concentrations in complicated pregnancies and also to explore the relationship between activin A levels and blood flow velocity in fetal arteries. Umbilical cord blood samples were obtained postpartum after a full term uneventful gestation (control group, n=40), and from pregnancies complicated by gestational diabetes (n=13), preterm labour (n=18), or pre-eclampsia (n=19). Cord serum activin A levels were three-fold higher in pregnancies complicated by pre-eclampsia (1.17+/-0.14 ng/ml, p<0.01) than in the control group (0.43+/-0.03 ng/ml), but were unaltered in the diabetes and preterm labour groups. The pre-eclampsia group had a marked increase of umbilical artery pulsatility index (PI) and also a decrease of middle cerebral artery PI (p<0.01). Furthermore, activin A concentration correlated directly with the umbilical artery PI (r=0.540, p=0.021), with the length of stay in the Neonatal Intensive Care Unit (r=0.857, p<0.001) and also with cord blood pH (r=-0.886, p<0.001). In conclusion, umbilical cord serum activin A levels are increased in the presence of pre-eclampsia and provide an indirect marker of impaired blood flow in the uteroplacental and fetal circulation.     

Transforming growth factor-beta 1 serum levels in pregnancy and pre-eclampsia

BACKGROUND: Isoforms of transforming growth factor-beta (TGF-beta1) are thought to be involved in the pathogenesis of pre-eclampsia. Data with respect to TGF-beta1 are controversial. We examined the correlation between TGF-beta1 serum levels and the occurrence and severity of pre-eclampsia. METHODS: Transforming growth factor-beta 1 serum levels were measured using a commercially available enzyme-linked immunosorbent assay in 44 women with pre-eclampsia and 44 healthy pregnant women. Results were correlated with clinical data. RESULTS: No difference in TGF-beta1 serum levels was assessed between women with pre-eclampsia and healthy pregnant women. Transforming growth factor-beta 1 serum levels were not associated with the severity of the disease and were not correlated with clinical maternal (blood pressure, proteinuria) and fetal (5-min APGAR score, umbilical cord pH values, birth weight) parameters. CONCLUSIONS: Our data support the assumption that in contrast to other isoforms, TGF-beta1, as evidenced by serum TGF-beta1 levels, does not seem to be involved in the pathogenesis of pre-eclampsia.     

Low levels of maternal serum PAPP-A in the first trimester and the risk of pre-eclampsia

BACKGROUND: Low levels of pregnancy associated plasma protein-A (PAPP-A) have been previously shown to be associated with pregnancies that subsequently develop pre-eclampsia. The objective of this study was to establish the relative risk for pre-eclampsia at various PAPP-A levels as an aid to counselling and follow up of pregnancies. METHODS: Maternal serum PAPP-A and free ss-human chorionic gonadotropin (ss-hCG) levels at 11 to 13 weeks of gestation from 224 singleton pregnancies that subsequently developed pre-eclampsia were compared to those from 47,770 normal singleton pregnancies resulting in live births after 37 weeks with birth weight greater than or equal to the 10th centile of normal for gestation. In all cases, the measured PAPP-A and free ss-hCG levels were expressed as multiple of the median (MoM). The association between metabolite levels and the incidence of pre-eclampsia was assessed by comparing the relative incidence at a number of MoM cut-offs and at various centiles. At various marker levels, the likelihood ratio for pre-eclampsia was also calculated. RESULTS: In the pre-eclampsia group the median PAPP-A MoM was significantly reduced (0.772 MoM, p < 0.0001) whilst the median free beta-hCG MoM was not different from controls (0.981 MoM, p = 0.26). With decreasing levels of PAPP-A, the likelihood ratio for pre-eclampsia increased. At the 5th centile of normal (PAPP-A MoM 0.415) the odds ratio was increased 4-fold and at this cut-off 15% of cases of pre-eclampsia would be identified. CONCLUSIONS: The graphical presentation of a likelihood ratio curve for pre-eclampsia at any PAPP-A MoM level is likely to be useful in counselling women with low levels of PAPP-A and a normal karyotype. Use of low levels of PAPP-A for selecting women for further follow-up with uterine artery Doppler may further improve the clinical discrimination.     

Galectin-7 serum levels are altered prior to the onset of pre-eclampsia

Galectins regulate many cell functions important for placental development, however, the localization and role of galectin-7 is unknown. We hypothesized galectin-7 would be expressed by the placenta and detected in serum. Galectin-7 immunolocalized to syncytiotrophoblast, extravillous trophoblast and glandular epithelium in 1st trimester placenta/decidua and to syncytiotrophoblast and endothelial cells in term placenta, but in pre-eclamptic placentas endothelial staining was absent. Galectin-7 serum concentration was significantly elevated in women (weeks 10–12 and 17–20) who subsequently developed pre-eclampsia compared to women with healthy pregnancies. Galectin-7 is a promising prospective serum biomarker for pre-eclampsia and likely has important functions in placentation.     

Biochemical markers of maternal bone turnover are elevated in pre-eclampsia

Objectives To investigate the hypothesis that bone turnover is reduced in pre-eclampsia compared with normal pregnancy.
Design A prospective sectional study.
Setting Obstetric unit at the Chelsea and Westminster Hospital, London.
Methods Third trimester maternal plasma levels of the linked carboxyl-terminal telo-peptide of type I collagen (ICTP), (a marker of bone resorption) and the carboxyl terminal pro-peptide of type I pro-collagen (PICP), (a marker of bone formation) were compared in 25 women with pre-eclampsia and in 24 normal pregnant controls. The subjects were matched for maternal age, booking body mass index and gestational age. PICP and ICTP levels were measured by radio-immunoassay.
Results ICTP and PICP levels were significantly increased in women with pre-eclampsia compared with the normal pregnant controls (   P = 0.0001 and P = 0.004, for ICTP and PICP respectively, Wilcoxon signed ranked test  ). There was no significant correlation between either of the markers and booking body mass index, blood pressure, serum uric acid levels or platelet count.
Conclusions These data suggest that bone turnover is increased in established pre-eclampsia compared with normal pregnancy. Further studies are required to investigate whether this precedes the onset of the disease.     

Smoking in pregnancy is associated with increased total maternal serum cell-free DNA levels

OBJECTIVE: Cell-free DNA is a marker of cellular apoptosis and necrosis. We wished to determine if maternal smoking affects maternal and fetal serum cell-free DNA levels. METHODS: Case-control sets of stored second-trimester serum-screening samples from 27 smoking and 90 nonsmoking pregnant women were developed. Smoking status was confirmed by measuring serum cotinine levels. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and DYS1 levels were determined using real-time polymerase chain reaction (PCR) to measure total and fetal cell-free DNA, respectively. At delivery, medical records were reviewed to confirm gender and determine other factors that could affect DNA values. RESULTS: Smoking was associated with significantly elevated GAPDH levels compared with nonsmokers (median: 97,662 genome equivalents (GE)/mL vs 38,217 GE/mL; p = 0.018). DYS1 levels were not statistically significantly elevated in smokers (p = 0.29). Other factors that affected DYS1 levels included maternal age in nonsmokers only (r(2) = 0.30, p = 0.013) and maternal Synthroid use (p = 0.0045) CONCLUSION: Pregnant smokers have threefold higher levels of total cell-free DNA compared with pregnant nonsmokers. Maternal age and Synthroid exposure may also affect circulating cell-free fetal DNA levels.     

Association between maternal serum 25-hydroxyvitamin D level and pre-eclampsia

Objectives: The objective of the study was to evaluate the association of maternal plasma levels of 25-hydroxyvitamin D (25(OH)D) at late second and third trimester and the risk of pre-eclampsia.

Methods: In this prospective cohort study, maternal plasma 25(OH)D levels were measured at late second and third trimester in 77 women who later developed pre-eclampsia (31 non-severe and 46 severe cases) and 180 women without pre-eclampsia.

Results: The mean gestational age of the timing of the blood sampling was 31.1?±?4.4 at control group, 32.6?±?5.7 at non-severe pre-eclamptic group and 32.3?±?5.4 at severe pre-eclamptic group. The mean 25(OH)D concentration was significantly low in severe pre-eclampsia group (5.8?±?4.5?ng/ml) than non-severe pre-eclampsia (11.8?±?7.3?ng/ml, p?=?0.039) and control groups (14.9?±?12.0?ng/ml, p?<?0.0001). There was no statistically significant difference in 25(OH)D concentration between non-severe pre-eclamptic and control groups (p?=?0.404). In women with 25(OH)D concentration <20?ng/ml, a 12.45-fold increase in the odds of severe pre-eclampsia were detected.

Conclusion: Women with severe pre-eclampsia had low serum 25(OH)D levels. The correlation between maternal 25(OH)D levels and aspartate aminotransferase, alanine transaminase, serum creatinine levels, platelet count were not determined. 25(OH)D levels may be used as an independent predictive marker of severe pre-eclampsia.     

Implications of 'low' maternal serum alpha-fetoprotein levels: are maternal age risk criteria obsolete?

An association has been reported between 'low' maternal serum alpha-fetoprotein (MSAFP) and fetal chromosome abnormalities, notably Down syndrome. We suggest the predictive value be used for genetic counselling when a 'low' MSAFP is found, and present an illustrative risk table. It can also be seen that normal MSAFP in a woman may lower her age-related risks below that previously defined as 'high risk'. However, until good estimates of sensitivity and specificity are available from prospective, population based studies, patients should be told that any risk estimates are rough approximations. When good estimates are available, use of age risks alone may become obsolete.     

Circulating concentrations of sFlt1 (soluble fms-like tyrosine kinase 1) in fetal and maternal serum during pre-eclampsia

OBJECTIVE: We hypothesized that umbilical vein serum soluble fms-like tyrosine kinase 1 (sFlt1) concentration was augmented in pre-eclampsia. We also explored a possible association between fetal and maternal concentrations of sFlt1. STUDY DESIGN: At cesarean delivery, maternal serum samples from pre-eclamptic (n=38) and uncomplicated (n=32) pregnancies were obtained, as well as umbilical vein serum and amniotic fluid samples. ELISA for human sFlt1, vascular endothelial growth factor (VEGF) and placental growth factor (PlGF) were performed. RESULTS: Median sFlt1 concentrations were elevated in pre-eclampsia compared to uncomplicated pregnancy, in umbilical venous serum (246 and 163 pg/mL, P=0.04), in maternal serum (9932 and 3417 pg/mL, P<0.001), as well as in amniotic fluid (51,040 and 33,490 pg/mL, P=0.03). A positive association between the fetal and maternal serum levels of sFlt1 was found in the pre-eclampsia group. Median PlGF concentration in the maternal serum was significantly lower in the pre-eclampsia group compared to the control group (82 pg/mL and 169 pg/mL, P<0.001). CONCLUSIONS: sFlt1 concentration is elevated in the fetal circulation in pre-eclampsia, but at a much lower level than in the maternal circulation. The results of our study do not support a substantial fetal contribution to the elevated circulating maternal sFlt1 protein concentration in pre-eclampsia.     

First-trimester maternal serum activin A in pre-eclampsia and fetal growth restriction.

OBJECTIVE: To investigate whether the reported increase in maternal serum activin A concentration in pre-eclampsia is evident from the first trimester. DESIGN: This was a case-control study carried out in antenatal clinics among singleton pregnancies at 10-14 weeks of gestation. METHODS: Activin A concentration was measured in stored maternal serum samples obtained at 11-14 weeks of gestation from 131 women who subsequently developed pre-eclampsia, 77 who developed non-proteinuric pregnancy-induced hypertension, 141 with fetal growth restriction in the absence of hypertensive complications and from 494 normotensive controls. RESULTS: Compared to the median activin A level in the control group (1.00 MoM), the median MoM in the patients who subsequently developed pre-eclampsia and pregnancy-induced hypertension (1.49 MoM and 1.32 MoM, respectively) was significantly increased (p < 0.001), and in patients with fetal growth restriction (1.02 MoM) it was not significantly different (p = 0.57). In the pre-eclampsia group (n = 131) the disease was considered to be sufficiently severe to necessitate iatrogenic delivery before 35 weeks in 25 patients, and in this group the median MoM was 1.92. CONCLUSION: Maternal serum activin A concentration at 12 weeks of gestation in pregnancies which subsequently develop hypertensive disease is increased, whereas in those complicated by fetal growth restriction it is normal.     

Serum levels of nesfatin-1 are increased in gestational diabetes mellitus

Objective: To analyze the concentrations of nesfatin-1 in maternal and cord serum, to evaluate the expression of nesfatin-1 in subcutaneous adipose tissue (SAT) from pregnant women with gestational diabetes mellitus (GDM) and those with normal glucose tolerance (NGT).

Methods: We studied a total of 50 GDM and 50 NGT subjects. The clinical features, serum nesfatin-1, homeostasis model assessment of insulin resistance (HOMA-IR), lipid profiles were measured at the third trimester of pregnancy. The expression of nesfatin-1 in the SAT was determined by western blot.

Results: Compared with the NGT group, the GDM group showed greater levels of serum nesfatin-1, adipocyte fatty acid binding protein (AFABP), and leptin; a greater level of cord blood nesfatin-1; and a higher level of expression in SAT (p?p?b?=?0.317, p=?0.022) and body mass index (BMI) before delivery (b?=?0.367, p=0.008) were independently associated with serum nesfatin-1. Nesfatin-1 was the independent risk factor for GDM.

Conclusions: The GDM group had higher levels of maternal serum and cord blood nesfatin-1, and greater nesfatin-1 expression in SAT. Nesfatin-1 is closely related to obesity and IR in pregnancy.     

Elevated maternal and fetal serum interleukin-6 levels are associated with epidural fever

OBJECTIVE: The study was performed to explore the evidence for a noninfectious inflammatory cause for epidural fever. STUDY DESIGN: A secondary analysis of a prospective randomized trial was performed. At epidural placement, afebrile term nulliparous women were randomized to receive acetaminophen 650 mg or placebo every 4 hours. Maternal serum was collected every 4 hours until delivery. Cord blood samples were collected at delivery. Interleukin-6 (IL-6) and interleukin-8 levels were measured using enzyme-linked immunosorbent assay techniques. Student t tests, chi(2), repeated measure analysis of variance, Pearson correlation coefficients, and linear regression modeling were used as appropriate. RESULTS: Twenty-one subjects received placebo, and 21 received acetaminophen. The rate of fever more than 100.4 degrees F was identical in the placebo and acetaminophen groups (23.8%). Maternal serum IL-6 levels before delivery were significantly higher in mothers who had a fever (596.0 vs 169.1 pg/mL, P <.001), as was the cord blood IL-6 levels of their infants (370.5 vs 99.0 pg/mL, P <.01). Linear regression modeling demonstrated that initial maternal serum IL-6, fever, and duration of epidural but not length of rupture of membranes or number of vaginal examinations were significantly associated with final maternal serum IL-6 levels. All neonatal blood cultures were negative. CONCLUSION: Epidural fever is associated with maternal and fetal inflammation in the absence of neonatal infection. Higher levels of cytokines in maternal serum suggest that the maternal compartment is the primary inflammatory source.     

子痫前期患者血清及胎盘组织中人类白细胞抗原G的表达研究

目的:检测正常晚期妊娠妇女及子痫前期患者胎盘组织HLA-G的表达及血清中sHLA-G的浓度,探讨HLA-G在子痫前期发病中的临床意义。方法:用半定量逆转录-聚合酶链技术(RT-PCR)检测37例正常晚期妊娠妇女(正常妊娠组)及41例子痫前期患者(轻度20例,重度21例)胎盘组织中HLA-G mRNA的表达;并用ELISA检测血清sHLA-G浓度。结果:(1)子痫前期胎盘组织中HLA-G mRNA表达水平分别为:轻度0.402±0.104、重度0.329±0.09,明显低于正常妊娠组的0.628±0.117(P<0.01),轻、重度差异亦有统计学意义(P<0.05)。(2)子痫前期血清sHLA-G浓度:轻度45.5±11.9u/ml,重度31.2±10.3u/ml,均明显低于正常妊娠组的105.7±12.5u/ml(P<0.01),轻重度差异亦有统计学意义(P<0.01)。(3)子痫前期患者血清sHLA-G浓度与胎盘组织HLA-G mRNA表达水平呈正相关(r=0.702,P<0.01)。结论:子痫前期血清sHLA-G浓度及胎盘组织中HLA-G表达水平均明显降低,可能与子痫前期发病及病情轻重程度相关。     

Visfatin serum levels are increased in women with preeclampsia: A case-control study

Objective: Visfatin has been implicated in the pathogenesis of preeclampsia with limited and contradictory, however, results. The aim of this study was to investigate the potential association between visfatin serum concentration and preeclampsia. Methods: Visfatin was determined in the serum of 38 women with preeclampsia and 38 women with uncomplicated pregnancies, matched for age and gestational age. Results: Similar baseline characteristics were present between the two groups in terms of age, body mass index, parity and gravidity. Serum visfatin was significantly increased in the preeclamptic women (median?=?10.3?ng/mL; interquartile range [IQR]?=?20) as opposed to their matched controls (median?=?2.6?ng/mL; IQR?=?1.4) (p?<?0.001). Univariate analysis revealed a strong linear correlation of visfatin levels with systolic (r?=?0.505, p?<?0.001), diastolic (r?=?0.467, p?<?0.001) and mean arterial blood pressure (r?=?0.497, p?<?0.001), as well as with uric acid concentrations in the serum (r?=?0.463, p?<?0.001). A receiver operating characteristics curve analysis illustrated that serum visfatin concentration is helpful in discriminating between preeclamptic or nonpreeclamptic women with an area under the curve of 0.887 (95% confidence interval [CI]: 0.794–0.948; p?<?0.001). Conclusion: Visfatin serum concentration seems to be increased in preeclampsia as compared with uncomplicated pregnancy.