Stephanie Worton

Analysis of the Metabolic Footprint and Tissue Metabolome of Placental Villous Explants Cultured at Different Oxygen Tensions Reveals Novel Redox Biomarkers

Pre-eclampsia (PE) is a multi-system disorder of pregnancy hypothesised to arise from circulating factors derived from an unhealthy placenta. Some changes in placental phenotype seen in PE can be reproduced by culture in altered oxygen (O2) tension. Currently, these circulating factors are unidentified, partly due to the complexity of maternal plasma. Investigation of factors released from placental tissue provides a potential method to identify bioactive compounds. Experimental strategies to study compounds present in a biological system have expanded greatly in recent years. Metabolomics can detect and identify endogenous and secreted metabolites. We aimed to determine whether metabolites could be identified in placental cultures with acceptable experimental variability and to determine whether altered O2 tension affects the composition of the placental metabolome. In this study we used gas-chromatography-mass spectroscopy to determine the presence of metabolites in conditioned culture medium (CCM) and tissue lysates of placental villous explants cultured in 1, 6 and 20% atmospheric O2 for 96h. This experimental strategy had an intra-assay variation of 6.1-11.6%. Intra and inter-placental variability were 15.7-35.8% and 44.8-46.2% respectively. Metabolic differences were identified between samples cultured in 1, 6 and 20% O2 in both CCM and tissue lysate. Differentially expressed metabolites included: 2-deoxyribose, threitol or erythritol and hexadecanoic acid. We conclude that metabolomic strategies offer a novel approach to investigate placental function. When conducted under carefully controlled conditions, with appropriate statistical analysis, metabolic differences can be identified in placental explants in response to altered O2 tension. Metabolomics could be used to identify changes in conditions associated with placental pathology.

IFPA meeting 2014 workshop report: Animal models to study pregnancy pathologies; new approaches to study human placental exposure to xenobiotics; biomarkers of pregnancy pathologies; placental genetics and epigenetics; the placenta and stillbirth and fetal growth restriction

Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialized topics. At IFPA meeting 2014 there were six themed workshops, five of which are summarized in this report. These workshops related to various aspects of placental biology but collectively covered areas of animal models, xenobiotics, pathological biomarkers, genetics and epigenetics, and stillbirth and fetal growth restriction.

UNDERSTANDING THE PLACENTAL AETIOLOGY OF FETAL GROWTH RESTRICTION; COULD THIS LEAD TO PERSONALIZED MANAGEMENT STRATEGIES?

Fetal growth restriction (FGR) is defined as the failure of a fetus to attain its full genetic growth potential. It is a leading cause of stillbirth, prematurity, cerebral palsy and perinatal mortality. Small size at birth increases surviving infants’ lifelong risk of adverse health outcomes associated with the metabolic syndrome. The pathophysiology of abnormal fetal growth is extremely complex and incompletely understood, with a plethora of genetic, signalling and metabolic candidates under investigation, many of which may result in abnormal structure and function of the placenta. In contrast to, or maybe because of, the underlying complexities of FGR, the strategies clinicians have for identifying and managing this outcome are conspicuously limited. Current clinical practice is restricted to identifying pregnancies at risk of FGR, and when FGR is detected, using intensive monitoring to guide the timing of delivery to optimise fetal outcomes. Abnormal Doppler indices in the umbilical artery are strongly associated with poor perinatal outcomes and are currently the “gold standard” for clinical surveillance of the growth-restricted fetus.

Altered Plasma Metabolome in Early Pregnancy in Women Who Develop a Small for Gestational Age Baby

Plenary Session: Trainee Plenary (Thursday, 3/19/2009, 9:00 AM 10:00 AM) Scientifi c Abstracts Reproductive Sciences Vol. 16, No. 3 (Supplement), March 2009 69ABackground: Small for Gestational Age (SGA) is defi ned as <10th centile on individualised birthweight centiles. SGA babies are at increased risk of complications in pregnancy and later life such as stillbirth, preterm delivery, cerebral palsy and heart disease and diabetes in adulthood. Metabolomics is the study of global metabolite profi les in a biological system. This technology has previously identifi ed differences in maternal plasma between normal and preeclamptic pregnancies. Aim: To examine the differences in the plasma metabolome in early pregnancy between women who develop a SGA baby and normal matched controls. Methods: 60 primigravid women who delivered a SGA baby were identifi ed within the SCOPE study (www.scopestudy.net) and matched to 60 controls. Venepuncture was performed at 15�1 weeks gestation. Samples were analysed using Ultra Performance Liquid Chromatography(UPLC)/LTQ-Orbitrap Mass Spectrometry (MS). Univariate statistical analysis was performed. Subsequently all the peaks in each class were grouped and analysed in combination by employing Multivariate Discriminant Analysis. Results: Univariate analysis of the UPLC/LTQ-Orbitrap MS data at 15 weeks gestation identifi ed 111 ?information rich? peaks (p < 0.05). The multivariate discriminant model (all 111 peaks), provided an area under the ROC curve of 0.96. Downloaded from rsx.sagepub.com at The John Rylands University Library, The University of Manchester on January 28, 2011Plenary Session: Trainee Plenary (Thursday, 3/19/2009, 9:00 AM 10:00 AM) Scienti c Abstracts Reproductive Sciences Vol. 16, No. 3 (Supplement), March 2009 69A

Metabolic Footprinting of Preeclamptic Placental Tissue Using UPLC/LTQ-Orbitrap Mass Spectrometry

Plenary Session: Trainee Plenary (Thursday, 3/19/2009, 9:00 AM 10:00 AM) Scientifi c Abstracts Reproductive Sciences Vol. 16, No. 3 (Supplement), March 2009 69ABackground: Small for Gestational Age (SGA) is defi ned as <10th centile on individualised birthweight centiles. SGA babies are at increased risk of complications in pregnancy and later life such as stillbirth, preterm delivery, cerebral palsy and heart disease and diabetes in adulthood. Metabolomics is the study of global metabolite profi les in a biological system. This technology has previously identifi ed differences in maternal plasma between normal and preeclamptic pregnancies. Aim: To examine the differences in the plasma metabolome in early pregnancy between women who develop a SGA baby and normal matched controls. Methods: 60 primigravid women who delivered a SGA baby were identifi ed within the SCOPE study (www.scopestudy.net) and matched to 60 controls. Venepuncture was performed at 15�1 weeks gestation. Samples were analysed using Ultra Performance Liquid Chromatography(UPLC)/LTQ-Orbitrap Mass Spectrometry (MS). Univariate statistical analysis was performed. Subsequently all the peaks in each class were grouped and analysed in combination by employing Multivariate Discriminant Analysis. Results: Univariate analysis of the UPLC/LTQ-Orbitrap MS data at 15 weeks gestation identifi ed 111 ?information rich? peaks (p < 0.05). The multivariate discriminant model (all 111 peaks), provided an area under the ROC curve of 0.96. Downloaded from rsx.sagepub.com at The John Rylands University Library, The University of Manchester on January 28, 2011Plenary Session: Trainee Plenary (Thursday, 3/19/2009, 9:00 AM 10:00 AM) Scienti c Abstracts Reproductive Sciences Vol. 16, No. 3 (Supplement), March 2009 69A