Maternal Placenta Research Articles

Profiling placental DNA methylation associated with maternal SSRI treatment during pregnancy

Selective serotonin reuptake inhibitors (SSRIs) for treatment of prenatal maternal depression have been associated with neonatal neurobehavioral disturbances, though the molecular mechanisms remain poorly understood. In utero exposure to SSRIs may affect DNA methylation (DNAme) in the human placenta, an epigenetic mark that is established during development and is associated with gene expression. Chorionic villus samples from 64 human placentas were profiled with the Illumina MethylationEPIC BeadChip; clinical assessments of maternal mood and SSRI treatment records were collected at multiple time points during pregnancy. Case distribution was 20 SSRI-exposed cases and 44 SSRI non-exposed cases. Maternal depression was defined using a mean maternal Hamilton Depression score > 8 to indicate symptomatic depressed mood (“maternally-depressed”), and we further classified cases into SSRI-exposed, maternally-depressed (n = 14); SSRI-exposed, not maternally-depressed (n = 6); SSRI non-exposed, maternally-depressed (n = 20); and SSRI non-exposed, not maternally-depressed (n = 24). For replication, Illumina 450K DNAme profiles were obtained from 34 additional cases from an independent cohort (n = 17 SSRI-exposed, n = 17 SSRI non-exposed). No CpGs were differentially methylated at FDR < 0.05 comparing SSRI-exposed to non-exposed placentas, in a model adjusted for mean maternal Hamilton Depression score, or in a model restricted to maternally-depressed cases with and without SSRI exposure. However, at a relaxed threshold of FDR < 0.25, five CpGs were differentially methylated (|Δβ| > 0.03) by SSRI exposure status. Four were covered by the replication cohort measured by the 450K array, but none replicated. No CpGs were differentially methylated (FDR < 0.25) comparing maternally depressed to not depressed cases. In sex-stratified analyses for SSRI-exposed versus non-exposed cases (females n = 31; males n = 33), three additional CpGs in females, but none in males, were differentially methylated at the relaxed FDR < 0.25 cut-off. We did not observe large-scale alterations of DNAme in placentas exposed to maternal SSRI treatment, as compared to placentas with no SSRI exposure. We also found no evidence for altered DNAme in maternal depression-exposed versus depression non-exposed placentas. This novel work in a prospectively-recruited cohort with clinician-ascertained SSRI exposure and mood assessments would benefit from future replication.

Influence of Collagen Production, Collagenase Activity and Amino Acids Level on Formation of Fetal Growth Restriction in Women with Placental Insufficiency

Rationale. According to WHO, about 21 million children are born annually in the world with low body weight. Currently, the problems of etiology, diagnosis and treatment of fetal growth retardation (FGR) are very relevant for obstetrics and perinatology. Purpose to study the level of collagen (COL1) collagenase activity (MMP1), and content of proline, ornithine in the maternal blood serum, amniotic fluid, placenta and cord blood of patients with placental insufficiency, and to identify their role in the formation of fetal growth restriction (FGR) during early pregnancy. Methods. 150 women were examined, which made up 2 groups control (with uncomplicated pregnancy, n = 105) and main (with IUGR, n = 45). All measurements were performed in maternal blood serum, amniotic fluid, placenta and cord blood of women of both groups. Collagen content and collagenase activity are determined by ELISA kits, USA. Determination of amino acids (proline, ornithine) was carried out by capillary electrophoresis on the device Capel-105 (St. Petersburg). Results. As a result of the correlation analysis in the study of all biofluids by the Spearman method with the calculation of the rank correlation coefficient (r) in the group of women with FGR, significant correlations between collagen production and collagenase activity were revealed. The most pronounced interdependence (r = 0.85; p 0.05) was found in the maternal blood serum. Conclusions. A relationship was found between the studied parameters (collagen, collagenase, proline, ornithine) in blood serum, amniotic fluid, cord blood and placenta of women with placental insufficiency and their role in the formation of fetal growth retardation was established.

Vascular Distribution and Expression Patterns of Angiogenic Factors in Caruncle during the Early Stage of Pregnancy in Goats (Capra hircus).

The placenta is a temporary maternal−fetal organ, and its maternal placenta (caruncle) is essential for fetal growth and development. The exchange function of the placenta requires vascular development (angiogenesis). However, the angiogenesis of the caruncle is poorly understood in goats during the early stage of pregnancy. Here, we investigated the vascular distribution, mRNA expression of major angiogenic factors, and the methylation levels of ANGPT2 in the goat caruncle. It showed that CAD (capillary area density), CSD (capillary surface density), and APC (area per capillary) increased gradually, while CND (capillary number density) showed an insignificant change, probably due to the variability between animals. The proportion of proliferating cells was observed to be very high (>26%) and increased (p < 0.002) approximately 2-fold from day 20 to 60 of pregnancy. Furthermore, the expression patterns of major angiogenic factors changed during the early stage of pregnancy. Interestingly, we discovered an absolute correlation between the mRNA for ANGPT2, TEK, FGF2, and vascular distribution. Subsequently, we evaluated the DNA methylation of ANGPT2, where we found that mean methylation was negatively correlated with CAD. The methylation at the CpG sites, such as CpG 4/18, CpG 9.10.11, and CpG 15, showed significant changes during the early stage of pregnancy. Thus, our findings suggest that the methylation of ANGPT2 may be involved in the regulation of caruncle angiogenesis during the early stage of pregnancy.

Role of Uterine Telocytes During Pregnancy

Abstract Endometrial remolding and angiogenesis are critical events that occur during pregnancy in order to establish uteroplacental vascular communication. This study investigated the role of uterine telocytes (TCs) in pregnancy. We analyzed the distribution of TCs and morphological changes in the endometrium of the gravid rabbit uterus at different stages of pregnancy: after ovulation, pre-implantation (day 7), post-implantation (days 8 and 9), and mid-pregnancy (day 14) and late (days 21–28) pregnancy. TCs gradually increased with the progression of pregnancy. They had distinctive telopodes (TPs) and podoms, with intranucleolar chromatin. The TCs established contact with decidual cells, growing a glandular epithelium, blood vessels, and immune cells, such as lymphocytes, neutrophils, and macrophages. The TCs underwent morphological changes at the post-implantation phase. They acquired thick and voluminous TPs, formed an extensive three-dimensional (3D) labyrinth at mid-pregnancy, and exhibited irregular-shaped nuclei and a dilated rough endoplasmic reticulum at late pregnancy. They also acquired a convoluted contour-formed complex network. Scanning electron microscopy (SEM) showed an extensive 3D network in the endometrium, forming a condensed sheath at late pregnancy. Transmission electron microscopy and SEM detected fenestrated TPs, and TCs were identified by CD34 and vascular endothelial growth factor expression. TCs also expressed matrix metalloproteinase-9 and transforming growth factor beta-1. Results suggested that TCs might play an essential role in maternal placenta formation, especially decidualization, regulation of uterine gland development, and neovascularization of maternal uterine blood vessels.

Distribution of endotoxin in maternal and fetal body with intrahepatic cholestasis of pregnancy and its association with adverse fetal outcome

Intrahepatic cholestasis of pregnancy is a pregnancy-specific liver disease. In this study, we sought to explore the distribution of lipopolysaccharide in the maternal body, and its effect on the fetal body in the intrahepatic cholestasis of pregnancy mice. It provides a new sight for the clinical treatment of women with intrahepatic cholestasis of pregnancy. The serum levels of lipopolysaccharide and lipopolysaccharide binding protein in women with intrahepatic cholestasis of pregnancy were analyzed. To assess the association between lipopolysaccharide levels and adverse fetal outcomes, ursodeoxycholic acid, resveratrol, and phosphatidylinositol-3-kinase inhibitor were employed in intrahepatic cholestasis of pregnancy mice, and we studied the fluorescence intensity and distribution of lipopolysaccharide in mice with intrahepatic cholestasis of pregnancy. Our data indicated significantly elevated levels of lipopolysaccharide and lipopolysaccharide binding protein in women with intrahepatic cholestasis of pregnancy. In vivo fluorescence imaging revealed that the intensity of lipopolysaccharide in mice with intrahepatic cholestasis of pregnancy was higher than that in the control group, and decreased after ursodeoxycholic and resveratrol treatment. The fluorescence intensity analysis indicated that lipopolysaccharide levels in maternal liver, placenta, fetal brain and fetal liver were significantly higher in the intrahepatic cholestasis pregnancy mice group than in the control group. This study provided evidence of endotoxin distribution in maternal liver, placenta, fetal liver and fetal brain in mice with intrahepatic cholestasis of pregnancy. Ursodeoxycholic acid and resveratrol treatment effectively reduced lipopolysaccharide levels in pregnant mice with intrahepatic cholestasis of pregnancy.

MORPHOLOGICAL AND IMMUNOHISTOCHEMICAL FEATURES OF PLACENTA AND PLACENTAL FACTOR IN WOMEN WITH ISOLATED CONGENITAL HEART DEFECTS IN NEWBORN

Introduction. Congenital heart defects (CHD) in fetus can affect the course of pregnancy, cause changes in the functioning of the pregnant woman's systems and metabolic and hemodynamic disorders. The relevance of the problems CHD is primarily due to their high frequency in newborns (5-9 per 1,000 live births) depending on geographical differences and the significant share among multiple congenital malformations and the need for surgical treatment in the neonatal period. Placenta, as one of the least studied human organs, plays not only a crucial role in the process of organogenesis, but also in the "programming" of human health after fetal and newborn periods. For today, it’s important to determine the association between the CHD and placental abnormalities, the nature and consequences of which need further study.Objective. To study morphological and immunohistochemical features of placental structures in parturients with isolated congenital heart defects in the newborn.Material and methods. Over the period 2020-2021, 90 pregnant women aged 17 to 39 years (mean age 28.36±5.08) who gave birth at 37-41 weeks, and their placents, of which 30 had different forms of CHD in the newborn, were examined on a hospital-based sample. Patients were divided into two groups: the study - women with CHD in the newborn, and control - somatically healthy motherswith healthy children. Given the different etiologies of cardiovascular defects in fetus, as well as the presence of different risk factorsfor their occurrence, in order to minimize their impact on the emergence of isolated CHD, considered appropriate selection criteria for pregnant women with non-syndromic forms of cardiovascular defects, as the presence of genetic syndrome can affect intrauterine fetaldevelopment, which makes it impossible to determine a single effect of CHD development. The selection criteria in this study included:somatically healthy woman, unburdened pregnancy, natural insemination (without using of reproductive technologies), the presence of isolated heart disease in the fetus, confirmed by a cardiologist using echocardiography. Organometric, macroscopic, general histological and immunohistochemical methods were used to study placental morphology and placental factors.Results. Analyzing the data on the morphological features of placentas from women with CHD in the newborn, we should note the presence of circulatory disorders in 60% of cases, namely multiple infarcts, complete or partial obliteration of arteries and arterioles with perivascular fibrosis (in the stem villi – 15%, intermediate – 10%, terminal - 10%), reduced number of fetal vessels in the terminal villiand syncytiocapillary membranes, abrupt vasodilation of intermediate villi. Complete or partial replacement of villous epithelium by fibrinoid masses in 20% was also detected. Immunohistochemical study of placental growth factor showed less pronounced expression in syncytia, decidual sheath and stromal cells (1-2 points) compared with control (2-3 points).Conclusions. The revealed morphological and immunohistochemical changes in the placental barrier at different levels refer to the damage of maternal and fetal placenta structures of parturients with isolated neonatal heart disease, which lead to impaired stromalvascular processes with reduced perfusion. This confirms the world data on the presence of placental-cardiac axis and can be a predictor of CHD at the early stages of placental structure study, as well as in further prediction of the child's health.

HAEMOLYTIC OF NEWBORN DISEASE-RELATED RED BLOOD CELL ALLOANTIBODIES

Red cell alloimmunisation in pregnancy may result in haemolytic disease of the foetus and newborn (HDN). During pregnancy, there are possibilities of the foetal antigen being exposed to the mother’s circulation, thereby leading to the production of antibodies. The immunoglobulin G (IgG) antibodies from the mother pass through the maternal surface placenta, which is then sensitised and obstructs the formation of foetal red blood cells (RBCs). This review discusses the presence of variance alloantibodies during pregnancy, as well as the consequences of HDN and its management. This review revealed that ABO incompatibility is the most common cause of alloimmunisation in pregnancy. The Rh system (anti-D and -E) are the most common alloantibodies found in pregnant women, followed by other alloantibodies such as Kell, Duffy, Kidd and MNSs. Hyperbilirunemia (increased bilirubin levels) is commonly seen in neonatal jaundice due to HDN and it is usually manageable using the phototherapy method. Nevertheless, the clinical significance of alloantibodies can cause complications such as anaemia, and in some extreme cases, kernicterus might require a blood exchange transfusion.

The effects of maternal cigarette smoking on cadmium and lead levels, miRNA expression and biochemical parameters across the feto-placental unit

Several miRNAs have been previously identified to be associated with cigarette smoke and/or the toxic metals cadmium (Cd) and lead (Pb). The aim of this study was to investigate the associations of maternal cigarette smoking with cadmium (Cd) and lead (Pb) levels, candidate miRNA expression and biochemical parameters across the feto-placental unit. miRNAs were isolated according to protocols provided by manufacturer from 72 healthy postpartum women using Qiagens’ kits based on phenol/guanidine samples lysis and silica-membrane purification of total RNA. Candidate miRNAs (miR-1537, miR-190b, miR-16, miR-21, and miR-146a) were quantified by real-time PCR. Biochemical parameters were analyzed in plasma samples by standardized and harmonized enzymatic methods using appropriate calibrators, while CRP was determined by immunoturbidimetric method. Concentration of Cd and Pb in whole blood and placenta samples were measured by inductively coupled plasma mass spectroscopy. Cd levels in smokers were higher in all of the analyzed compartments of the feto-placental unit, Pb in maternal blood and placenta than non-smokers. Smokers also had a higher expression of miR-16 in maternal and miR-146a in cord plasma, and lower expression of miR-21 in the placenta in comparison to non-smokers. Urate concentrations in the maternal plasma of smokers were lower than this value in non-smokers. The study has demonstrated that maternal smoking was associated with toxic metals (Cd and Pb) levels, urate concentration and alteration of miRNA expression. Given that the effects of maternal smoking on miRNA expression are inadequate, all compartments of the feto-placental unit should be analyzed to obtain a complete picture. This paper is the first to report on the results of expression of cellular and circulating miRNAs simultaneously in maternal and fetal compartments and in the placenta.

Characterisation of cytochrome P450 isoenzyme activity in sheep liver and placental microsomes

IntroductionDrug metabolism during pregnancy is a complex process that involves maternal, placental and fetal sites of metabolism. Indeed, there is a lack of clarity provided from drug metabolism in human pregnancy due to ethical limitations. Large animal models of human pregnancy provide an opportunity to quantify activity of phase 1 drug metabolism mediated by cytochrome P450 (CYP) enzymes in the maternal, placental, and fetal compartments. Herein, we have validated a comprehensive assay to quantify maternal, placental, and fetal CYP activity. MethodsIsolated microsomes from sheep maternal liver, placenta, and fetal liver (140d gestation, term = 150d) were incubated with CYP-specific probe drugs to quantify the activity of CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A. Inhibition studies were performed to validate specificity of probe drugs. The validated assay was developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). ResultsCYP1A2, CYP2B6, CYP2C8, CYP2C19, CYP2D6, CYP2E1 and CYP3A were active in maternal liver. In contrast, only CYP1A2, CYP2C8 and CYP2D6 were active in the placenta, whereas CYP2B6, CYP2C8 and CYP2D6 were active in the fetal liver. Of the placental-specific CYPs validated, CYP1A2 increased in type A compared with type D placentomes, whereas CYP2C8 activity increased in type B compared with type A and C. DiscussionThis study has established conditions for compartment-specific CYP activity in the sheep maternal-placental-fetal unit using a validated and standardised experimental workflow. Compartment- and placentome type-specific CYP activity are important considerations when examining drug metabolism in the maternal-placental-fetal unit and in determining the impact of pregnancy complications.

Placental DAAM2 is unaltered in preeclampsia, but upregulated by treatment with proton pump inhibitors.

Dishevelled Associated Activator Of Morphogenesis 2 (DAAM2) levels are elevated in the maternal circulation and placenta in pregnancies complicated by fetal growth restriction. However, placental DAAM2 levels in cases of preeclampsia have not previously been explored. Here, we examined placental DAAM2 in pregnancies complicated by preterm preeclampsia, and whether candidate preeclampsia therapeutics altered its expression. DAAM2 mRNA and protein levels were assessed in placental tissue from cases of preterm preeclampsia and gestation-matched controls (delivering ≤ 34weeks; qPCR and western blot respectively). Short interfering RNAs were used to silence DAAM2 in isolated primary cytotrophoblast under normoxic (8% O2) and hypoxic (1% O2) conditions, and expression of anti-angiogenic sFLT-1, angiogenic PGF, antioxidant, fetal growth, and inflammatory genes assessed. DAAM2 expression was measured in placental explant tissue from pregnancies complicated by preeclampsia, treated with three proton pump inhibitors (100µM esomeprazole, lansoprazole, and rabeprazole). DAAM2 expression was significantly reduced in preeclamptic placental tissue compared to controls, but protein production was unchanged. Silencing DAAM2 in hypoxic cytotrophoblast increased sFLT-i13 isoform expression, but did not alter sFLT-e15a or PGF expression, or sFLT-1 secretion. DAAM2 knockdown did not alter expression of antioxidant (NQO-1, TXN, GCLC), fetal growth (SPINT1), or inflammasome (NLRP3) genes. Esomeprazole and lansoprazole, but not rabeprazole, increased DAAM2 expression in placental explant tissue from cases of preeclampsia. Placental DAAM2 protein is not significantly altered in placental tissue in cases of preeclampsia, and its suppression does not alter sFLT-1 secretion. Hence, placental DAAM2 is unlikely to drive the pathogenesis associated with preeclampsia.

Maternal and Fetal Exposure to (-)-Δ9-tetrahydrocannabinol and Its Major Metabolites in Pregnant Mice Is Differentially Impacted by P-glycoprotein and Breast Cancer Resistance Protein.

(-)-Δ9-tetrahydrocannabinol (THC) is the primary pharmacological active constituent of cannabis. 11-hydroxy-THC (11-OH-THC) and 11-nor-9-carboxy-THC (THC-COOH) are respectively the active and nonactive circulating metabolites of THC in humans. While previous animal studies reported that THC could be a substrate of mouse P-glycoprotein (P-gp) and breast cancer resistance protein (Bcrp), we have shown, in vitro, that only THC-COOH is a weak substrate of human BCRP, but not of P-gp. To confirm these findings and to investigate the role of P-gp and/or Bcrp in the maternal-fetal disposition of THC and its metabolites, we administrated 3 mg/kg of THC retro-orbitally to FVB wild-type (WT), P-gp -/-, Bcrp -/-, or P-gp-/- /Bcrp-/- pregnant mice on gestation day 18 and estimated the area under the concentration-time curve (AUC) of the cannabinoids in the maternal plasma, maternal brain, placenta, and fetus, as well as the tissue/maternal plasma AUC geometric mean ratios (GMRs) using a pooled data bootstrap approach. We found that the dose-normalized maternal plasma AUCs of THC in P-gp-/- and P-gp-/- /Bcrp-/- mice, and the placenta-to-maternal plasma AUC GMR of THC in Bcrp-/- mice were 279%, 271%, and 167% of those in WT mice, respectively. Surprisingly, the tissue-to-maternal plasma AUC GMRs of THC and its major metabolites in the maternal brain, placenta, or fetus in P-gp -/-, Bcrp -/- or P-gp-/- /Bcrp-/- mice were 28-78% of those in WT mice. This study revealed that P-gp and Bcrp do not play a role in limiting maternal brain and fetal exposure to THC and its major metabolites in pregnant mice. SIGNIFICANCE STATEMENT: This study systematically investigated whether P-gp and/or Bcrp in pregnant mice can alter the disposition of THC, 11-OH-THC, and THC-COOH. Surprisingly, except for Bcrp, which limits placental (but not fetal) exposure to THC, we found that P-gp-/- , Bcrp-/- , and/or P-gp-/- /Bcrp-/- significantly decreased exposure to THC and/or its metabolites in maternal brain, placenta, or fetus. The mechanistic basis for this decrease is unclear and needs further investigation. If replicated in humans, P-gp- or BCRP-based drug-cannabinoid interactions are not of concern.

The new landscape of differentially expression proteins in placenta tissues of gestational diabetes based on iTRAQ proteomics

IntroductionGestational diabetes mellitus (GDM) refers to abnormal glucose tolerance that occurs or is firstly diagnosed during pregnancy. GDM is related to various adverse pregnancy outcomes, but GDM pathogeny has not been fully elucidated. Nevertheless, previous studies have observed that many proteins in the placentas of patients with GDM are dysregulated. The present study aimed to establish a novel differentially expressed protein (DEP) landscape of GDM and normal maternal placentas and to explore the possible connection between DEPs and GDM pathogenesis. This study provides new insights into the mechanism of GDM and should make an important contribution to the development of biomarkers. MethodsThe morphological characteristics of the placenta were observed on 30 GDM and normal maternal placental tissues stained with haematoxylin and eosin. Isobaric tags for relative and absolute quantitation (iTRAQ) was used in the proteomics screening of the DEPs of the normal and GDM maternal placentas. Bioinformatics analysis was performed on the DEPs, and parallel reaction monitoring (PRM) was performed to verify the DEPs. Finally, the quantitative analysis of iTRAQ and PRM was verified by immunohistochemical assay. ResultsA total of 68 DEPs in the GDM placenta were identified with iTRAQ proteomics experiment, comprising 21 up-regulated and 47 down-regulated DEPs. Bioinformatics analysis showed that the regulation of transport, catabolic process of non-coding RNA, cytoskeleton and cell binding were the most abundant Gene Ontology terms, and RNA degradation was an important pathway for significant enrichment. Protein–protein interaction network analysis showed that heterogeneous nuclear ribonucleoproteins A2/B1 (HNRNPA2B1), heterogeneous nuclear ribonucleoprotein A/B (HNRNPAB), heterogeneous nuclear ribonucleoprotein L (HNRNPL) and heterogeneous nuclear ribonucleoprotein A3 (HNRNPA3) were the cores of the up-regulated proteins. Band 3 anion transport protein (SLC4A1), spectrin beta chain erythrocytic (SPTB), ankyrin-1 (ANK1), spectrin beta chain non-erythrocytic 2 (SPTBN2), D-3-phosphoglycerate dehydrogenase (PHGDH) and exosome complex component RRP42 (EXOSC7) were the cores of the down-regulated proteins. These proteins are involved in the binding, splicing, processing, transport and degradation of RNA and in the formation and maintenance of the cytoskeleton. PRM verification results showed that seven proteins, namely, epiplakin (EPPK1), cold-inducible RNA-binding protein (CIRBP), HNRNPA2B1, HNRNPAB, HNRNPL, Ras-related protein Rab-21 (RAB21) and Ras-related protein Rab-3B (RAB3B), were up-regulated, whereas SPTB and SLC4A1 were down-regulated. The results of immunohistochemical assay also showed that the expression of five proteins, namely EPPK1, HNRNPA2B1, HNRNPAB, CIRBP and RAB21, were significantly higher in GDM placental tissues (P < 0.01). The GDM placentas showed changes in the morphological evaluation, including poor villous maturation, obvious increase in the number of syncytiotrophoblast nodules, thickening of the wall of dry villous arterioles with lumen stenosis, increased fibrinous exudation and excessive filling of villous interstitial vessels. DiscussionDifferentially expressed proteins related to a variety of biological processes in the GDM placenta were found. Fourteen proteins, namely, HNRNPA2B1, HNRNPAB, HNRNPL, HNRNPA3, EPPK1, CIRBP, RAB21, RAB3B, SLC4A1, SPTB, ANK1, SPTBN2, PHGDH and EXOSC7, which were differentially expressed in the placenta, may play an important role in regulating the occurrence and development of gestational diabetes through multi-channel and multi-link regulation.

Influence of PRRSV-1 vaccination and infection on mononuclear immune cells at the maternal-fetal interface.

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most devastating viruses for the global swine industry. Infection during late gestation causes reproductive failure but the local immune response in utero remains poorly understood. In this study, an experimental PRRSV-infection model with two different PRRSV-1 field isolates was used to investigate the immune cell phenotypes at the maternal-fetal interface during late gestation. In addition, phenotypic changes induced by a modified live virus (MLV, ReproCyc® PRRS EU) vaccine were studied. Vaccinated (n = 12) and non-vaccinated pregnant gilts (n = 12) were challenged with either one of the PRRSV-1 field isolates (low vs. high virulent, LV or HV) or sham-inoculated at day 84 of gestation. Twenty-one days post infection all gilts were euthanized and the fetal preservation status for all fetuses per litter was assessed. Leukocytes from the maternal-fetal interface were isolated and PRRSV-induced changes were investigated using ex vivo phenotyping by flow cytometry. PRRSV load in tissue from the maternal endometrium (ME) and fetal placenta (FP) was determined by RT-qPCR. In the ME, a vast increase in CD8β T cells with CD8αposCD27dim early effector phenotype was found for fetuses from the non-vaccinated LV and HV-challenged gilts, compared to non-treated and vaccinated-only controls. HV-challenged fetuses also showed significant increases of lymphocytes with effector phenotypes in the FP, including NKp46pos NK cells, CD8αhigh γδ T cells, as well as CD8αposCD27pos/dim CD4 and CD8 T cells. In vaccinated animals, this common activation of effector phenotypes was more confined and the fetal preservation status significantly improved. Furthermore, a negative correlation between the viral load and CD163highCD169pos mononuclear phagocytic cells was observed in the FP of HV-infected animals. These results suggest that the strong expansion of effector lymphocytes in gilts that were only infected causes immune-pathogenesis rather than protection. In contrast, the attenuated MLV seems to dampen this effect, yet presumably induces memory cells that limit reproductive failure. This work provides valuable insights into changes of local immune cell phenotypes following PRRSV vaccination and infection.

Murine influenza A virus disrupts immune profiles in the maternal gut and placenta during acute infection in a dose-dependent manner

Murine influenza A virus disrupts immune profiles in the maternal gut and placenta during acute infection in a dose-dependent manner

ODP321 Impact of a Maternal High-Sucrose Diet in Rats on Fetal Physiology and the Placenta

Abstract Consumption of sucrose (table sugar) is high in much of the world. The effects of maternal sucrose intake on the placenta and fetal brain remain unknown. In rats, maternal consumption of sucrose at a human-relevant level alters the mother's physiology and steroids, as well as the adult offspring's brain and behavior. Effects in mothers are impaired glucose tolerance, increased liver lipids, increased adipose inflammation, and decreased corticosterone levels in the blood but not in the brain. In contrast, in adult female offspring, maternal sucrose intake increases corticosterone levels in the blood and the brain. In adult male offspring, maternal sucrose intake increases preference for high-sucrose and high-fat diets as well as motivation for sugar rewards in a progressive ratio task. In this study, we investigate the underlying mechanisms of the observed behavioral and endocrine effects in the adult offspring. We examine anti-inflammatory steroids, steroidogenic enzymes and cytokines in the placenta, amniotic fluid, fetal blood and brain. In our model, we feed rat dams either a high-sucrose diet (26% of kCal) or an isocaloric, matched, control diet (1% sucrose) 10 weeks prior to and during gestation. At embryonic day 19 (E19), we collected maternal serum, placenta, amniotic fluid, fetal blood, and fetal brain. We used Palkovits punch to microdissect the placenta and fetal brain. Next, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS), which is highly sensitive and specific, to measure multiple steroids (e. g. corticosterone, estrone, allopregnanolone). We examined multiple regions of the fetal brain (e. g. prefrontal cortex, nucleus accumbens, hypothalamus, hippocampus). We will also examine neuronal proliferation, microglia and tyrosine hydroxylase immunoreactivity in the fetal brain. Maternal high-sucrose diet increased 11-dehydrocorticosterone (inactive metabolite of corticosterone) and aldosterone in maternal serum and amniotic fluid. Testosterone and androstenedione were significantly higher in the amniotic fluid of male fetuses than female fetuses. Placental steroidogenic enzymes 3β-HSD and CYP19 were not affected by maternal diet or fetal sex; however, 3β-HSD activity was higher in the decidua than in the fetal part of the placenta. Steroid and cytokine data from the placenta, fetal blood and fetal brain are currently being analyzed. Ongoing analyses examine how a maternal high-sucrose diet affects brain development possibly by increasing inflammation. Presentation: No date and time listed

Maternal exposure to ambient PM2.5 perturbs the metabolic homeostasis of maternal serum and placenta in mice

Epidemiological and animal studies have shown that maternal fine particulate matters (PM2.5) exposure correlates with various adverse pregnancy outcomes such as low birth weight (LBW) of offspring. However, the underlying biological mechanisms have not been fully understood. In this study, female C57Bl/6 J mice were exposed to filtered air (FA) or concentrated ambient PM2.5 (CAP) during pregestational and gestational periods, and metabolomics was performed to analyze the metabolic features in maternal serum and placenta by liquid chromatography-mass spectrometry (LC-MS). The partial least squares discriminate analysis (PLS-DA) displayed evident clustering of FA- and CAP-exposed samples for both maternal serum and placenta. In addition, pathway analysis identified that vitamin digestion and absorption was perturbed in maternal serum, while metabolic pathways including arachidonic acid metabolism, serotonergic synapse, 2-oxocarboxylic acid metabolism and cAMP signaling pathway were perturbed in placenta. Further analysis indicated that CAP exposure influenced the nutrient transportation capacity of placenta, by not only changing the ratios of some critical metabolites in placenta to maternal serum but also significantly altering the expressions of nutrition transporters in placenta. These findings reaffirm the importance of protecting women from PM2.5 exposure, and also advance our understanding of the toxic actions of ambient PM2.5.

PS-2 Early Intrauterine Suppression of CXCL12-CXCR4 Alters Expression of Placental Inflammatory Profile at Mid-to-Late Gestation

Abstract A proper balance of inflammatory cytokines at the fetal-maternal interface is crucial for successful embryo implantation and placental formation. Chemokine ligand 12 (CXCL12) and receptor CXCR4 are expressed by fetal and maternal placental tissues and implicated in cytokine synthesis. However, mechanisms controlling the axis and placental inflammatory profile are not well characterized. Using an in-vivo sheep model, we previously demonstrated suppressing CXCL12-CXCR4 signaling at the fetal-maternal interface during implantation yielded dysregulated production of inflammatory cytokines in the placenta. Our objective was to determine if disrupting the cytokine profile during implantation alters the placental inflammatory profile at mid-to-late gestation. Day 12 post-breeding, osmotic pumps were surgically installed in 37 ewes to deliver CXCR4 inhibitor (1.5X dose, n = 8; 1X dose, n = 8; 3X dose, n = 8) or saline (n = 13) into the uterine lumen ipsilateral to the corpus luteum for 14 days. On days 90 and 135 of gestation, placental tissues were collected; maternal (caruncle) and fetal (cotyledon) placenta components were then separated and analyzed. Real-time qPCR was used to measure gene expression of pro-inflammatory cytokines tumor necrosis factor (TNF), interferon-γ (IFNγ), interleukin-12 (IL12), and anti-inflammatory cytokines interleukin-10 (IL10) and transforming growth factor-β (TGFβ). Gene expression for TGFβ from tissues ipsilateral to the pump on day 90 increased (P &amp;lt; 0.03) in caruncles from 3x treated ewes compared to control ewes while IL10 mRNA decreased (P &amp;lt; 0.02) in cotyledons from 1.5x compared to 3x treatment. Furthermore, analysis of day 135 1x treated ewes showed greater expression (P &amp;lt; 0.03) of IL12 in caruncles ipsilateral compared to contralateral of the pump; meanwhile IL10 showed decreased expression (P &amp;lt; 0.02) in cotyledons ipsilateral compared to contralateral of the pump. High levels of TGFβ, IFNγ (whose production is promoted by increased IL12), and TNF along with decreased levels of IL10 have been associated with preeclampsia in women. The changes in gene expressions of ovine immune cytokines with a disrupted CXCL12-CCR4 axis may be related to preeclamptic events. Ultimately, modulating CXCL12-induced actions is a novel approach to manipulating the fetal-maternal environment when most pregnancy losses occur and may reveal methods to improve reproductive success and fetal health in livestock.

PSIII-1 Late-Breaking: Fighting Fetal Morbidity at the Fetal-Maternal Interface One Chemokine Access at a Time

Abstract Improper placentation is the root cause to intrauterine growth restriction (IUGR) and fetal mortality in livestock. This study aims to further characterize the importance of the C-X-C motif chemokine receptor 4 (CXCR4) and its ligand CXCL12 on placentation and fetal growth through potential impacts on the insulin-like growth factors (IGF) and their binding proteins (IGFBP1-3). CXCR4 and CXCL12 are implicated in placentation though the exact roles are unclear. The peptides insulin-like growth factors 1 and 2 have impacts on fetal growth, placental development, and trophoblast differentiation and if the IGFs are down regulated, a corresponding decrease in placenta and fetus size has been observed. The roles of the IGFs and their binding proteins in disease states like IUGR are not completely known but studies have demonstrated them to be contributing factors. Using an in-vivo sheep model we surgically inserted osmotic pumps on day 12 (d12) of gestation ipsilateral to the corpus luteum to deliver treatments into the uterus of bred ewes. Pumps delivered either saline (control) or AMD3100, the CXCR4 inhibitor at a 1x, 1.5x, or 3x dose into the uterine lumen over the course of 14 days. Our objective was to determine if disrupting the CXCL12/CXCR4 axis during implantation alters the expression of IGFs and their binding proteins in the placenta at mid and late gestation. At midgestation (d90) and late gestation (d135) placental and fetal samples were obtained for analysis. Of the targets tested, mRNA expression was significantly decreased by AMD3100 treatment for both IGF-1 (P = 0.05) and IGF-2 (P = 0.02) compared to control primarily in maternal placenta tissues on d135. For fetal tissue samples, d90 mRNA expression for IGF-1 (P = 0.04) IGFBP-2 (P = 0.02) and IGFBP-3 (P = 0.03) increased in the 3x AMD3100 treated group compared to the control while in d135 samples, IGF-1was significantly decreased (P = 0.04) by the 1x treatments compared to control. Our preliminary data show an importance of the CXCL12- CXCR4 signaling on placentation and that impairment of this axis may alter IGFs in multiple tissue types later in gestation. Insights into this chemokine axis will lead to a greater understanding of placental development and its importance to the health of mother and offspring.

Alterations in maternal plasma exosomal miRNAs revealed selective material exchange between maternal circulation and placenta.

Exosomes have emerged as important regulators in the communication between maternal peripheral blood and placenta. We aimed to compare maternal plasma exosomal miRNAs profile between healthy pregnant and nonpregnant women, screen for differential expressed miRNAs and their potential regulatory role during pregnancy. We isolated exosomes from plasma of mid-trimester, last trimester, and nonpregnant women (n=6 each group), analyzed the miRNA profile using next-generation sequencing. Several miRNA clusters were expressed in plasma exosomes, such as C19MC, C14MC, and let-7 family, miRNAs in each cluster may have synergistic effect during pregnancy. We assumed maternal circulating exosomal miRNA could be transported into placenta or selectively uptook by placenta, which was consistent with the fact that many pregnancy-associated or placenta highly expressed miRNAs reduced in exosomes during pregnancy. Some exosomal miRNAs were mainly secreted by the placenta, which could act as markers that reflect changes in the function and microenvironment of the placenta. Exosomal miRNAs are associated with placenta development and have potential as molecular markers.

Maternal cadmium exposure impairs placental angiogenesis in preeclampsia through disturbing thyroid hormone receptor signaling.

Cadmium is a ubiquitous environmental pollutant, which can increase the risk of preeclampsia. This study was designed to determine the mechanism of cadmium exposure during pregnancy impaired placental angiogenesis that was associated with the occurrence of preeclampsia. The effects of cadmium exposure on placental thyroid hormone receptor signaling were explored. JEG3 cells were treated with CdCl2 (20μM) and the Dio2 inhibitor, IOP (100μM). Cadmium levels in maternal blood and placentae were increased in preeclampsia group. Placental angiogenesis of preeclampsia was decreased with decreased expression of PLGF and VEGF and increased expression of sFlt1. Meanwhile, the expression and nuclear translocation of thyroid hormone receptor α were decreased in preeclampsia placenta, as well as the expression of Dio2, but not the expression and nuclear translocation of thyroid hormone receptor β. Furthermore, we found that cadmium exposure downregulated the expression of thyroid hormone receptor α and Dio2, but not the expression of thyroid hormone receptor β in JEG3 cells. Also, we found that cadmium exposure decreased the expression of PLGF and VEGF and increased the expression of sFlt1 in JEG3 cells. IOP pretreatment decreased the expression of PLGF and increased the expression of sFlt1. In conclusion, our results elucidated that cadmium exposure would impair placental angiogenesis in preeclampsia through disturbing thyroid hormone receptor signaling.