Pathogenesis Of Early-onset Preeclampsia Research Articles

GPR124 promotes trophoblast proliferation, migration, and invasion and inhibits trophoblast cell apoptosis and inflammation via JNK and P38 MAPK pathways.

Early-onset preeclampsia, which occurrs before 34 weeks of gestation, is the most dangerous classification of preeclampsia, which is a pregnancy-specific disease that causes 1% of maternal deaths. G protein-coupled receptor 124 (GPR124) is significantly expressed at various stages of the human reproductive process, particularly during embryogenesis and angiogenesis. Our prior investigation demonstrated a notable decrease in GPR124 expression in the placentas of patients with early-onset preeclampsia compared to that in normal pregnancy placentas. However, there is a lack of extensive investigation into the molecular processes that contribute to the role of GPR124 in placenta development. This study aimed to examine the mechanisms by which GPR124 affects the occurrence of early-onset preeclampsia and its function in trophoblast. Proliferative, invasive, migratory, apoptotic, and inflammatory processes were identified in GPR124 knockdown, GPR124 overexpression, and normal HTR8/SVneo cells. The mechanism of GPR124-mediated cell function in GPR124 knockdown HTR8/SVneo cells was examined using inhibitors of the JNK or P38 MAPK pathway. Downregulation of GPR124 was found to significantly inhibit proliferation, invasion and migration, and promote apoptosis of HTR8/SVneo cells when compared to the control and GPR124 overexpression groups. This observation is consistent with the pathological characteristics of preeclampsia. In addition, GPR124 overexpression inhibits the secretion of pro-inflammatory cytokines interleukin (IL)-8 and interferon-γ (IFN-γ) while enhancing the secretion of the anti-inflammatory cytokine interleukin (IL)-4. Furthermore, GPR124 suppresses the activation of P-JNK and P-P38 within the JNK/P38 MAPK pathway. The invasion, apoptosis, and inflammation mediated by GPR124 were partially restored by suppressing the JNK and P38 MAPK pathways in HTR8/SVneo cells. GPR124 plays a crucial role in regulating trophoblast proliferation, invasion, migration, apoptosis, and inflammation via the JNK and P38 MAPK pathways. Furthermore, the effect of GPR124 on trophoblast suggests its involvement in the pathogenesis of early-onset preeclampsia.

Identification of the role of DAB2 and CXCL8 in uterine spiral artery remodeling in early-onset preeclampsia

Aberrant remodeling of uterine spiral arteries (SPA) is strongly associated with the pathogenesis of early-onset preeclampsia (EOPE). However, the complexities of SPA transformation remain inadequately understood. We conducted a single-cell RNA sequencing analysis of whole placental tissues derived from patients with EOPE and their corresponding controls, identified DAB2 as a key gene of interest and explored the mechanism underlying the communication between Extravillous trophoblast cells (EVTs) and decidual vascular smooth muscle cells (dVSMC) through cell models and a placenta-decidua coculture (PDC) model in vitro. DAB2 enhanced the motility and viability of HTR-8/SVneo cells. After exposure to conditioned medium (CM) from HTR-8/SVneoshNC cells, hVSMCs exhibited a rounded morphology, indicative of dedifferentiation, while CM-HTR-8/SVneoshDAB2 cells displayed a spindle-like morphology. Furthermore, the PDC model demonstrated that CM-HTR-8/SVneoshDAB2 was less conducive to vascular remodeling. Further in-depth mechanistic investigations revealed that C-X-C motif chemokine ligand 8 (CXCL8, also known as IL8) is a pivotal regulator governing the dedifferentiation of dVSMC. DAB2 expression in EVTs is critical for orchestrating the phenotypic transition and motility of dVSMC. These processes may be intricately linked to the CXCL8/PI3K/AKT pathway, underscoring its central role in intricate SPA remodeling.Graphical abstract

Bioinformatic Analysis Identifies Potential Extracellular Matrix Related Genes in the Pathogenesis of Early Onset Preeclampsia.

Early-onset preeclampsia (EOPE) is a complex pregnancy complication that poses significant risks to the health of both mothers and fetuses, and research on its pathogenesis and pathophysiology remains insuffcient. This study aims to explore the role of candidate genes and their potential interaction mechanisms in EOPE through bioinformatics analysis techniques. Two gene expression datasets, GSE44711 and GSE74341, were obtained from the Gene Expression Omnibus (GEO) to identify differentially expressed genes (DEGs) between EOPE and gestational age-matched preterm control samples. Functional enrichment analysis was performed utilizing the kyoto encyclopedia of genes and genomes (KEGG), gene ontology (GO), and gene set enrichment analysis (GSEA). A protein-protein interaction (PPI) network was constructed using the STRING database, and hub DEGs were identified through Cytoscape software and comparative toxicogenomics database (CTD) analysis. Furthermore, a diagnostic logistic model was established using these hub genes, which were confirmed through reverse transcription polymerase chain reaction (RT-PCR). Finally, immune cell infiltration was analyzed using CIBERSORT. In total, 807 DEGs were identified in the GSE44711 dataset (451 upregulated genes and 356 downregulated genes), and 787 DEGs were identified in the GSE74341 dataset (446 upregulated genes and 341 downregulated genes). These DEGs were significantly enriched in various molecular functions such as extracellular matrix structural constituent, receptor-ligand activity binding, cytokine activity, and platelet-derived growth factor. KEGG and GSEA annotation revealed significant enrichment in pathways related to ECM-receptor interaction, PI3K-AKT signaling, and focal adhesion. Ten hub genes were identified through the CytoHubba plugin in Cytoscape. Among these hub genes, three key DEGs (COL1A1, SPP1, and THY1) were selected using CTD analysis and various topological methods in Cytoscape. The diagnostic logistic model based on these three genes exhibited high efficiency in predicting EOPE (AUC = 0.922). RT-PCR analysis confirmed the downregulation of these genes in EOPE, and immune cell infiltration analysis suggested the significant role of M1 and M2 macrophages in EOPE. In conclusion, this study highlights the association of three key genes (COL1A1, SPP1, and THY1) with EOPE and their contribution to high diagnostic efficiency in the logistic model. Additionally, it provides new insights for future research on EOPE and emphasizes the diagnostic value of these identified genes. More research is needed to explore their functional and diagnostic significance in EOPE.

Downregulation of CLDN1 impairs trophoblast invasion and endovascular trophoblast differentiation in early-onset preeclampsia

IntroductionTo investigate the role of claudin-1 (CLDN1) in trophoblast invasion and endovascular trophoblast (enEVT) differentiation in early-onset preeclampsia (EOPE). MethodsThe expression and localization of CLDN1 in normal (n = 18) and EOPE (n = 20) placental tissues were detected by immunohistochemical (IHC) staining, quantitative real-time polymerase chain reaction (qRT‒PCR) and Western blotting. Next, invasion, migration and tube formation assays were performed to explore the involvement of CLDN1 in trophoblast invasion and enEVT differentiation in trophoblast cell lines (HTR8/SVneo). Then, invasion and enEVT markers were analyzed via Western blotting and qRT‒PCR, respectively. Finally, we established an EOPE mouse model to detect the Cldn1 protein level. ResultsCLDN1 expression was significantly decreased in EOPE placental tissues. Knockdown of CLDN1 suppressed HTR8/SVneo cell invasion, migration and the ability to penetrate the endothelial tube. Conversely, overexpression of CLDN1 promoted trophoblast invasion and the ability to invade the endothelial tube. Inhibition of CLDN1 decreased the protein expression of VIM and SNAIL along with downregulating IL1B and PECAM1 mRNA levels, while overexpression of CLDN1 gave the opposite results. In the EOPE mouse model, we found a decrease in Cldn1 expression in EOPE mouse placentas. DiscussionThese results suggest that the downregulation of CLDN1 in trophoblast cells is involved in the pathogenesis of early-onset preeclampsia by affecting trophoblast invasion and enEVT differentiation.

Kisspeptin Is Upregulated at the Maternal-Fetal Interface of the Preeclamptic-like BPH/5 Mouse and Normalized after Synchronization of Sex Steroid Hormones.

Insufficient invasion of conceptus-derived trophoblast cells in the maternal decidua is a key event in the development of early-onset preeclampsia (PE), a subtype of PE associated with high maternal and fetal morbidity and mortality. Kisspeptins, a family of peptides previously shown to inhibit trophoblast cell invasion, have been implicated in the pathogenesis of early-onset PE. However, a role of kisspeptin signaling during the genesis of this syndrome has not been elucidated. Herein, we used the preeclamptic-like BPH/5 mouse model to investigate kisspeptin expression and potential upstream regulatory mechanisms in a PE-like syndrome. Expression of the kisspeptin encoding gene, Kiss1, and the 10-amino-acid kisspeptide (Kp-10), are upregulated in the non-pregnant uterus of BPH/5 females during diestrus and in the maternal-fetal interface during embryonic implantation and decidualization. Correspondingly, the dysregulation of molecular pathways downstream to kisspeptins also occurs in this mouse model. BPH/5 females have abnormal sex steroid hormone profiles during early gestation. In this study, the normalization of circulating concentrations of 17β-estradiol (E2) and progesterone (P4) in pregnant BPH/5 females not only mitigated Kiss1 upregulation, but also rescued the expression of multiple molecules downstream to kisspeptin and ameliorated adverse fetoplacental outcomes. Those findings suggest that uterine Kiss1 upregulation occurs pre-pregnancy and persists during early gestation in a PE-like mouse model. Moreover, this study highlights the role of sex steroid hormones in uteroplacental Kiss1 dysregulation and the improvement of placentation by normalization of E2, P4 and Kiss1.

Artificial Synchronization of Sex Steroid Hormones Normalizes Kisspeptin Expression and Placentation in the Preeclamptic‐Like BPH/5 Mouse

Insufficient invasion of the maternal decidua by conceptus‐derived trophoblast cells is a key event in the pathogenesis of early‐onset preeclampsia (ePE), a subclass of PE associated with high maternal morbidity and fetal intrauterine growth restriction (IUGR). Kisspeptins (KP) are a family of small peptides previously shown to inhibit trophoblast cell invasion. Interestingly, KP expression is higher in the placentae of women with ePE than in those with uncomplicated pregnancies. Nonetheless, the mediators of placental KP upregulation remain largely unknown. The Blood Pressure High Subline 5 (BPH/5) is a mouse model that spontaneously develops the main features of PE, including decreased trophoblast cell invasion, lower decidual placental expansion, reduced placental labyrinth endothelial remodelling and IUGR. We have previously shown that Kiss1is higher at the BPH/5 maternal‐fetal interface during early gestation, at embryonic days (e) 4.5 and 7.5, when compared with control C57 mice. Moreover, BPH/5 females have abnormal sex steroid hormone profile during early gestation, including an earlier and depressed surge of estradiol‐17ß (E2) and higher blood progesterone (P4) at e2.5. Upregulation of uterine Kiss1expression has been previously demonstrated by administration of E2 and P4 to non‐pregnant ovariectomized CD‐1 mice. Therefore, we aimed to investigate the effects of artificial synchronization of sex steroid hormones (AS‐SSH) in BPH/5 uteroplacental Kiss1expression and placental morphology. It was hypothesized that AS‐SSH during early gestation would prevent the BPH/5 Kiss1upregulation at the embryonic implantation sites (eIS) and result in higher decidual placental expansion. AS‐SSH was performed via ovariectomy of pregnant BPH/5 and C57 females (n = 8/strain) at e2.5; administration of a single dose of E2 (25 ng/mouse, subcutaneously) to stimulate embryonic implantation and daily subcutaneous injections of P4 (1mg/mouse) until sample collection. The eIS were collected at e5.5 and Kiss1 expression was assessed via RT‐PCR. Additionally, e12.5 fetoplacental units from BPH/5 and C57 natural (Nat) and AS‐SSH pregnancies were stained with Isolectin, and the placental decidual expansion was assessed using Image J® (n = 4‐6/group). The placental expansion into the maternal decidua was calculated as the ratio of the labyrinth and junctional zone in relation to the entire placental disc (placenta + decidua). Comparisons were done using unpaired T‐tests and ANOVA with post‐hoc Newman‐Keuls tests. Contrary to Nat BPH/5 pregnancies, eIS Kiss1 expression was not different between AS‐SSH BPH/5 and C57 (p = 0.77). Nat BPH/5 had lower placental expansion (p < 0.05) than Nat C57, whereas AS‐SSH BPH/5 presented higher placental expansion than Nat BPH/5 (p < 0.05). In conclusion, normalization of the E2 and P4 profile in the PE‐like BPH/5 mouse early gestation not only mitigated the Kiss1upregulation at the maternal‐fetal interface, but also ameliorated the placental defects previously reported in this mouse model.

Expression profiles and functions of ferroptosis-related genes in the placental tissue samples of early- and late-onset preeclampsia patients

BackgroundThe accumulation of reactive oxygen species (ROS) resulting from upregulated levels of oxidative stress is commonly implicated in preeclampsia (PE). Ferroptosis is a novel form of iron-dependent cell death instigated by lipid peroxidation that likely plays an important role in PE pathogenesis. This study aimed to investigate the expression profiles and functions of ferroptosis-related genes (FRGs) in early-onset preeclampsia (EOPE) and late-onset preeclampsia (LOPE).MethodsGene expression data and clinical information were downloaded from the Gene Expression Omnibus (GEO) database. The “limma” R package was used to screen differentially expressed genes. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein–protein interaction (PPI) network analyses were conducted to investigate the bioinformatics functions and molecular interactions of significantly different FRGs. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to verify the expression of hub FRGs in PE.ResultsA total of 4215 differentially expressed genes (DEGs) were identified between EOPE and preterm cases while 556 DEGs were found between LOPE and term controls. Twenty significantly different FRGs were identified in EOPE subtypes, while only 3 FRGs were identified in LOPE subtypes. Functional enrichment analysis revealed that the differentially expressed FRGs were mainly involved in EOPE and enriched in hypoxia- and iron-related pathways, such as the response to hypoxia, iron homeostasis and iron ion binding process. PPI network analysis and verification by RT-qPCR resulted in the identification of the following five FRGs of interest: FTH1, HIF1A, FTL, MAPK8 and PLIN2.ConclusionsEOPE and LOPE have distinct underlying molecular mechanisms, and ferroptosis may be mainly implicated in the pathogenesis of EOPE. Further studies are necessary for deeper inquiry into placental ferroptosis and its role in the pathogenesis of EOPE.

Role of matrix metalloproteinase-9 expression in the pathogenesis of early-onset preeclampsia: a case–control study

Role of matrix metalloproteinase-9 expression in the pathogenesis of early-onset preeclampsia: a case–control study

Investigating the Roles of Small STBEV’s Micrornas in the Pathogenesis of Early-Onset Preeclampsia through Bioinformatics.

Investigating the Roles of Small STBEV’s Micrornas in the Pathogenesis of Early-Onset Preeclampsia through Bioinformatics.

418 Role of miRNA-452 expression in the pathogenesis of early onset preeclampsia: a case-control study

To investigate the placental tissues expression changes of miRNA-452 in patients with Early onset preeclampsia (EOPE). A case control study included 50 pregnant women scheduled for termination of pregnancy at gestational age (28-36) weeks was conducted between January 2019 and April 2020. Women were divided into two groups: severe EOPE group (n = 30) and healthy pregnant control group (n = 20). The diagnosis of PE is based on modified American College of Obstetricians and Gynecologists (ACOG) criteria 2014. A villus tissue was cut off immediately from the center of maternal side of the placenta, avoiding the area of infarction, bleeding or calcification. After being washed with normal saline, the tissue was preserved for subsequent detection of miR-452 expression by real time quantitative PCR (r-qPCR). Continuous data were expressed as medians. Differences between the two groups were detected using Mann-Whitney test. Nominal data were expressed as percentage, differences between the two groups were detected using Chi square test. Spearman’s correlation between placental weight and relative expression of placental miRNA-452 was used. Both groups were comparable as regard their age, body-mass index, gestational age and mode of delivery. Neonatal and placental weights were significantly higher in control group (p<0.001). miRNA-452 was significantly higher in the placentae of the PE group compared to that of the control group (7.49 vs. 1.07, p <0.001). Moreover, placental miRNA-452 had a significant negative correlation with the placental weight (r= -0.305, p <0.031) In conclusion, miRNA-452 expression in the placenta was significantly upregulated in patients with early onset PE compared to healthy pregnant women.

A Potential Role for CXCR2 in Early-onset Preeclampsia: Placental CXCR2 Expression is Related to Increased Blood Pressure and Serum LDH Levels

Abstract Objective: This study was aimed to determine the changes in CXCR2 expression in preeclampsia placenta and its correlation with clinical parameters. Methods: Sixty-four gravidas ranging in age from 25 to 42 years referred to the obstetrics unit of the West China Second University Hospital from April 2012 to October 2012 were recruited in this case-control study; women were diagnosed and divided into early-onset preeclampsia group (n = 22), late-onset preeclampsia group (n = 22), and healthy pregnancy group (n = 20). After immunolocalized in human placenta, the levels of CXCR2 protein and messenger ribonucleic acid (mRNA) were detected by enzyme-linked immunosorbent assay and real-time quantitative polymerase chain reaction. Correlations between placental CXCR2 protein expression with systolic blood pressure and lactate dehydrogenase (LDH) in early-onset preeclampsia were examined using Pearson or Spearman's correlation coefficients. Results: Placental CXCR2 protein and mRNA expression in early-onset preeclampsia was significantly lower than it was in placentas from healthy pregnancy pregnancies and late-onset preeclampsia (P &lt; 0.05). The placental CXCR2 protein expression correlated negatively with systolic blood pressure and LDH in early-onset preeclampsia (r = −0.51, P &lt; 0.05; r = −0.43, P &lt; 0.05). Conclusion: Significant abnormal placental CXCR2 expression in early-onset preeclampsia, and its correlations with some clinical parameters (systolic blood pressure and LDH) were discovered, suggesting that CXCR2 may play a role in the pathogenesis of early-onset preeclampsia.

Decreased circulating levels of plasmacytoid dendritic cells in women with early-onset preeclampsia

The role of plasmacytoid dendritic cells (pDCs), and myeloid dendritic cells (mDCs) in women with preeclampsia has not been elucidated. We compared the frequency of peripheral pDCs, mDCs, NK cells, and T helper 17 (Th17) cells among non-pregnant/pregnant women, and women with early-/late-onset preeclampsia. We examined pDCs and mDCs using Anti-Human Lineage Cocktail 3 (CD3, CD14, CD19, and CD20), HLA-DR, CD11c, and CD123. We detected NK cells using Lineage cocktail, CD8, CD16, and CD56. We determined Th17 cells using CD3, CD4, CD8, CXCR3, and CCR6. We recruited 13 non-pregnant women, 50 normal pregnant women, 13 women with early-onset preeclampsia (onset at <34 gestational weeks), and 10 women with late-onset preeclampsia. The fraction of pDCs in women with early-onset preeclampsia was significantly lower than in non-pregnant women and normal pregnant women at 19−29 gestational weeks (4.1 % vs. 41.2 % and 19.0 %, respectively [p = 0.0005, and p = 0.025]), however, the fraction of pDCs in late-onset preeclampsia was not significantly different from normal pregnant women at 37 gestational weeks (11.1 % vs. 29.1 %, respectively [p = 0.149]), although it was significantly lower than in non-pregnant women (11.1 % vs. 41.2 %, respectively [p = 0.044]). The fraction of Th17 cells in women with early-onset preeclampsia was significantly higher than in normal pregnant women at 19−29 gestational weeks (p = 0.022). In conclusion, the level of circulating pDCs was lower in early-onset preeclampsia than in non-pregnant and pregnant women, suggesting the role of pDCs in the pathogenesis of early-onset preeclampsia.

Investigation of mucosal-associated invariant T (MAIT) cells expressing immune checkpoint receptors (TIGIT and CD226) in early-onset preeclampsia

ObjectiveDuring our work, we examined the possible contribution of MAIT cells in the pathogenesis of the clinical phase of early-onset preeclampsia and how this could be influenced by TIGIT and CD226 immune checkpoint molecules. Study Design37 pregnant women diagnosed with early-onset preeclampsia and 36 healthy, age-matched control women were involved in this study. Peripheral blood mononuclear cells were isolated by density gradient and frozen. After thawing, cells were stained with monoclonal antibodies to characterize MAIT, MAIT-like, and non-MAIT cells. Flow cytometric analyses were used to measure TIGIT, CD226, intracellular perforin, and granzyme B expression. ResultsMAIT (CD3+ CD8+ Vα7.2+ CD161++), MAIT-like (CD3+ CD8+ Vα7.2+ CD161+) and non-MAIT (CD3+ CD8+ Vα7.2+ CD161-) cell population were identified based on their CD161 receptor positivity. MAIT cells markedly differed in proportion, TIGIT expression, granzyme B, and perforin content compared to MAIT-like and non-MAIT cells. A significant difference was determined in TIGIT expression by non-MAIT cells and in CD8/CD226 positive relationship between the preeclamptic and healthy condition. ConclusionsConsidering that we did not detect a notable difference between early-onset preeclampsia and healthy pregnancy, we suppose that peripheral MAIT cells expressing TIGIT and CD226 immune checkpoint molecules have a marginal role in the pathogenesis of early-onset preeclampsia.

Reduced ELABELA expression attenuates trophoblast invasion through the PI3K/AKT/mTOR pathway in early onset preeclampsia

IntroductionEarly onset preeclampsia is linked to abnormal trophoblast invasion, leading to insufficient recasting of uterine spiral arteries and shallow placental implantation. This study investigated ELABELA (ELA) expression and its involvement in the pathogenesis of early onset preeclampsia. MethodsWe used immunohistochemistry, quantitative PCR and Western blot to calculate ELA levels in the placentas. Transwell assays were utilize to assess the invasion and migration of trophoblastic Cells. Western blot was used to identify the concentrations of vital kinases in PI3K/AKT/mTOR pathways and invasion-related proteins in trophoblast cells. ResultsELA was expressed in villous cytotrophoblasts and syncytiotrophoblasts in placental tissue. Compared with the normal pregnancies, ELA mRNA and protein expression was significantly reduced in early onset preeclampsia placentas. In the HTR-8/SVneo cells, when ELA was knocked down, the invasion and migration capability of cells decreased significantly, with MMP2 and MMP9 expression downregulated and the expression of important kinases in the PI3K/AKT/mTOR pathways being significantly decreased compared to the control group. Overexpression of ELA was on the contrary. Besides, while PI3K was blocked, the invasion and migration capability of HTR-8/SVneo cells and the expression of key kinases in PI3K/AKT/mTOR pathways were decreased significantly. DiscussionELA stimulates the invasion and migration of trophoblastic cells through activation of downstream PI3K/AKT/mTOR pathway and is complicit in early onset preeclampsia pathogenesis. Our research offers a potential novel treatment for PE.

Correlation between the expression of STOX1 in placenta of patients with early onset preeclampsia

Objective: To investigate the relationship of STOX1expression and pathogenesis of early onset preeclampsia. Methods: 65 cases of preeclampsia women who delivered in Shanghai Pudong Hospital from October 2015 to June 2018, were recruited, which included 31 cases with early onset preeclampsia (early onset group, gestational week<34 weeks) and 34 patients with late onset preeclampsia (late onset group, gestational week ≥34 weeks). 34 cases women who received caesarean section because of pelvic structural deformities, breech presentation, macrosomia and social factors were included as the control group(gestational week ≥34 weeks) were selected as control group.The expression and localization of STOX1 mRNA and protein in placenta of three groups of maternal were evaluated by immunohistochemistry SP, RT-qPCR and Western blotting. Results: (1) The expression of STOX1 in placenta mainly distributed in the cytoplasm of placental syncytiotrophoblasts, cytotrophoblasts, vascular endothelial and mesenchymal cells, a few in the cell nucleus.The staining intensity of STOX1 in early onset group was significantly stronger than that in late onset group, the staining intensity of the late onset group was similar to that of the control group. The positive expression rates of STOX1protein in early onset group, late onset group and control group were 96.8%(30/31), 70.6%(24/34), 67.6%(23/34) respectively, which was higher in early onset group than that in late onset group(P=0.005). There was no statistical difference of STOX1 level between the late onset group and the control group(P=0.793). (2)Relative expression of STOX1 mRNA in early onset group, late onset group and control group were 0.054 3±0.003 5,0.037 5±0.000 7,0.035 2±0.000 4 respectively, which was significantly higher in early onset group than that in late onset group(P<0.05), while there was no statistical difference between the late onset group and the control group(P>0.05).(3)Relative expression level of STOX1 protein in early onset group, late onset group and control group were 0.78±0.04,0.59±0.020 and 0.54±0.018 respectively, which is higher in early onset group than that in late onset group(P<0.05). There was no statistical difference of STOX1 level between the late onset group and the control group(P>0.05). Conclusions: The pathogenesis of early onset and late onset preeclampsia may be different. Up-regulated expression of STOX1 in placenta may be associated with the pathogenesis of early onset preeclampsia.

Serum Calcium and 25-Hydroxy Vitamin D Level in Normal and Early Onset Pre-eclamptic Pregnant Women: A Study from Indonesia

Introduction: Pre-eclampsia is one of the most common causes of fetomaternal morbidity and mortality worldwide. This disorder is categorised into Early Onset Pre-eclampsia (EOPE) and Late-Onset Pre-eclampsia (LOPE). EOPE is usually accompanied by severe complications for both the mother and fetus, while LOPE is accompanied by relatively mild fetomaternal complications. Although the pathogenesis of EOPE is not yet fully elucidated, recent studies indicate that serum calcium and 25 Hydroxy Vitamin D (25(OH)D) levels may play a role in its pathogenesis. Aim: To find out the relationship of calcium and 25(OH)D serum levels in pregnant women with normal pregnancy and with EOPE in Indonesia. Materials and Methods: This study was a case-control study, conducted in Dr. Soetomo General Hospital from July to October 2017. A total of 36 women with EOPE and 64 women with normal pregnancy were included in this study. Inclusion criteria were pregnant women in 2nd or 3rd-trimester with BMI > 18 kg/m(2). Blood sample analysis was done to measure serum calcium and 25(OH)D level. Data were expressed as Mean +/- Standard Deviation. Data distribution was analysed using Shappiro-Wilk test. Comparison of serum calcium and 25(OH)D level between groups was analysed using Independent t-test. Correlation between serum calcium and 25(OH)D level was analysed using Pearson's correlation test. The p-value of Results: There was a significant difference in serum calcium level between case and control group (8.294 +/- 0.725 vs 8.670 +/- 0.405 mg/dL; p=0.006). In 25(OH)D level, there was no difference between both groups (16.128 +/- 7.5463 vs 17.325 +/- 6.4992 ng/mL; p=0.406). No correlation was found between calcium and 25(OH)D level (r=0.165; p=0.101). Conclusion: Calcium deficiency plays a role in the incidence of EOPE among pregnant women in Indonesia. The actual role of calcium deficiency in EOPE needs further investigation.

Altered DNA methylation and transcription of WNT2 and DKK1 genes in placentas associated with early-onset preeclampsia

PurposeThe purpose of this study was to characterize the changes in DNA methylation and transcription of WNT2 and DKK1 genes in placentas associated with early-onset preeclampsia. MethodsThe study includes three groups: patients with early-onset preeclampsia, normotensive preterm and term births. Placental tissues were collected and pyrosequencing was performed on DKK1 and WNT2 proximal promoters. Transcriptional levels of DKK1 and WNT2 genes were determined with real-time PCR. ResultsDKK1 gene methylation levels were lower in placentas associated with early-onset preeclampsia compared to those associated with preterm birth (P<0.05). DKK1 mRNA expression was higher in early-onset preeclampsia placentas than those in preterm placentas (P < 0.05). WNT2 mRNA expression in early-onset preeclamptic placentas was lower than that in other two groups (P < 0.05). In the preterm and early-onset preeclampsia groups, the mRNA levels for WNT2 and DKK1 were negatively correlated (P < 0.05). In all the subjects, the levels of DKK1 mRNA and methyaltion were negatively correlated (P < 0.05). ConclusionDecreased methylation of DKK1 promoter in early-onset preeclamptic placenta tissues may up-regulate the expression of DKK1. The increased expression of DKK1 and decreased expression of WNT2 may be involved in the pathogenesis of early-onset preeclampsia.

The possible role of CD8+/Vα7.2+/CD161++ T (MAIT) and CD8+/Vα7.2+/CD161lo T (MAIT-like) cells in the pathogenesis of early-onset pre-eclampsia.

The objective of this study was to compare the expressions of different immune-checkpoint molecules by MAIT and MAIT-like cells in healthy pregnancy and in early-onset pre-eclampsia. Peripheral blood mononuclear cells (PBMC) were stained with monoclonal antibodies to characterize MAIT and MAIT-like cells. Flow cytometric analyses were used to measure PD-1, TIM-3, activation markers, and intracellular perforin expression. We identified CD3+/CD8+/Vα7.2+/CD161++ MAIT cells and a minor cell population characterized by CD3+/CD8+/Vα7.2+/CD161lo surface markers. In measuring the expression of PD-1 receptor, we found a significantly lower expression by MAIT cells in women with early-onset pre-eclampsia. CD69 expression by MAIT cells was significantly elevated in early-onset pre-eclamptic patients. Intracellular perforin content by MAIT and PD-1+ MAIT cells was significantly increased in pre-eclamptic patients compared with healthy individuals. Altered frequency and reduced PD-1 expression combined together with the elevated perforin content of MAIT cells insinuate their potential roles in the pathogenesis of early-onset pre-eclampsia.

Distinct expression profiles of lncRNAs between early-onset preeclampsia and preterm controls

Early-onset preeclampsia (EOPE), which is the most severe form of the syndrome, confers a high risk of neonatal morbidity and perinatal death. We aim to study the roles of long non-coding RNAs (lncRNAs) in the pathogenesis of early-onset preeclampsia (EOPE). Therefore, we examined the expression profiles of lncRNAs between early-onset preeclampsia and preterm controls using microarray analysis. Quantitative real-time PCR (qRT-PCR) was performed to verify the selected differentially expressed lncRNAs. In total, we identified 15,646 upregulated and 13,178 downregulated lncRNAs in the placenta of EOPE patients compared to the preterm controls. Gene ontology and pathway analysis revealed that compared to the preterm controls, many of the processes over-represented in the EOPE patients were related to cell migration and cell motility. A selection of the differentially expressed lncRNA transcripts was confirmed using qRT-PCR, particularly RP11-465L10.10, which is associated with the MMP9 gene. These data may offer a background/reference resource for future functional studies of lncRNAs related to EOPE.

Interleukin 10 gene promoter polymorphisms in women with early-onset pre-eclampsia.

Pre-eclampsia is one of the most serious disorders of human pregnancy and T helper type 1 (Th1)/Th2 imbalance plays a major role in its aetiology. The Th2 cytokine, interleukin (IL)-10, plays a significant role in the maintenance of pregnancy. The present study is aimed at understanding the role of IL-10 promoter polymorphisms (-1082 G/A; -592 A/C and -819 C/T) and their haplotypes in early-onset pre-eclampsia. A total of 120 patients and an equal number of women with normal pregnancy, from Government Maternity Hospital, Petlaburz, Hyderabad, India, were considered for the present study. A standard amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) was carried out for genotyping followed by agarose gel electrophoresis. Appropriate statistical methods were applied to test for the significance of the results. It was found that the IL-10 -819 C allele (P = 0·003) and -592 A (P = 0·005) allele frequencies increased significantly in patients compared to controls. No significant difference was found with regard to -1082 promoter polymorphism. Haplotype analysis of the IL-10 single nucleotide polymorphisms (SNPs) revealed a significant association with ACC haplotype with a twofold increased risk in patients compared to controls. The frequencies of two common IL-10 haplotypes (GCC and ATA) did not show any significant difference. Further, the diplotype analysis revealed five genotypes: -1082A with -819C (P = 0·0016); -1082G with -819C (P = 0·0018); -819C with -592C (P = 0·001); -1082A with -592C (P = 0·032); and -1082G with -592C (P = 0·005) associated with the disease. These findings support the concept of contribution of IL-10 gene polymorphisms in the pathogenesis of early-onset pre-eclampsia.