The MEG3 lncRNA promotes trophoblastic cell growth and invasiveness in preeclampsia by acting as a sponge for miR-21, which regulates BMPR2 levels

Preeclampsia (PE) is one of the leading causes of maternal morbidity and mortality in pregnant women. This study aimed to investigate the potential impact and regulatory mechanisms of bone morphogenetic protein receptor 2 (BMPR2) on the progression of PE. We obtained placental tissues from pregnant women with PE and normal pregnant women, and the results showed that BMPR2 was expressed at low levels in the tissue from PE women. Genetic knockdown of BMPR2 increased the proliferation and invasion of cultured trophoblast cells, whereas its overexpression reduced these characteristics. Bioinformatics analysis and luciferase reporter gene assays confirmed that BMPR2 is a direct target of miR-21. Overexpression of a miR-21 inhibitor promoted the growth and invasiveness of trophoblast cells, whereas the opposite results were observed for the miR-21 mimic. Furthermore, miR-21 was sponged by the lncRNA MEG3, and shRNA inhibition of MEG3 reduced trophoblast cell growth and invasiveness. miR-21 was upregulated in the tissues from PE women, whereas MEG3 was downregulated, and the two were negatively correlated. Collectively, this study demonstrates that the lncRNA MEG3 acts as a sponge for miR-21, which regulates BMPR2 expression and promotes trophoblast cell proliferation and invasiveness, thereby preventing the development of PE. These findings provide novel insight into a targeted therapy that could be used to treat or prevent the development of PE.

Plasma SerpinA5 in conjunction with uterine artery pulsatility index and clinical risk factor for the early prediction of preeclampsia

Object This study aimed to combine plasma protein SerpinA5 with uterine artery doppler ultrasound and clinical risk factor during the first trimester for prediction of preeclampsia. Methods and materials This study was a nested cohort study and was divided into the screening set and developing set. The plasma was collected during the first trimester (11+0–13+6 weeks), at the same time, UtA-PI was detected and recorded with four-dimensional color Doppler ultrasound. These pregnancies were followed up until after delivery. The plasma proteins were examined using ultra-performance liquid chromatography–mass spectrometry (UPLC-MS) and enzyme linked immunosorbent assay (ELISA). Placental samples preserved after delivery were analysed by immunohistochemistry. Clinical risk factors were obtained from medical records or antenatal questionnaires. Upregulation or downregulation of SerpinA5 expression in TEV-1 cells was performed to investigate the role of SerpinA5 in trophoblasts invasion. Results We demonstrated that SerpinA5 levels were greater not only in preeclampsia placental tissue but also in plasma (both p<0.05), and we found that SerpinA5 may interfere with trophoblastic cell invasion by inhibiting MSP. SerpinA5 may be a potential predictor of preeclampsia. What is more, the sensitivity and specificity of predictive power were strengthened when plasma SerpinA5 was combined with UtA-PI and pre-pregnancy BMI & family history of PE for prediction of preeclampsia. Conclusion These findings showed that placenta-derived plasma SerpinA5 may be a novel biomarker for preeclampsia, which together with uterine artery Doppler ultrasound and clinical risk factor can more effectively predict preeclampsia.

Bone Marrow-Derived Mesenchymal Stem Cells (BMSCs)-Derived MicroRNA-378a-3p (miR-378a-3p) Inhibits the Migration of Gestational Trophoblast Cells and Epithelial Mesenchymal Transition via Regulating X-Linked Inhibitor of Apoptosis Protein (XIAP) Pathway

The components of the in vivo microenvironment are BMSCs and miRNAs that have a critical role in the development of pregnancy. Our aim was to further investigate the effect of the miRNAs of BMSC origin on pregnancy injury. Exosomal miR-378a-3p secreted by BMSCs was identified by electron microscopy and miR-378a-3p expression was measured during gestational injury. Target scan detects the correlation of XIAP and miR-378a-3p which was confirmed by luciferase activity along with analysis of cell growth by MTT assay and cell invasion by Transwell and EMT expression. Exosomal miR-378a-3p derived from BMSCs promoted proliferation and migration and invasion of trophoblast. miR-378a-3p targeted XIAP and its overexpression could significantly increase EMT switching. The miR-378a-3p/XIAP axis is critical in trophoblastic cell migration and EMT and is involved in pregnancy injury progression, indicating that it might be a novel potential target for the treatment of pregnancy injury.

Effect of Blastocyst Quality on Human Sex Ratio at Birth in Single Blastocyst Frozen Thawed Embryo Transfer Cycle

ObjectiveThis study aimed to determine whether blastocyst quality affect on sex ratio at birth through in single blastocyst frozen thawed embryo transfer cycle.Materials and MethodsWe ran this retrospective study on 4205 singleton infants born following single blastocyst frozen thawed embryo transfer from January 2016 to October 2020 at a single institution. We compared the sex ratios of these infants with respect to blastocyst quality, embryo growth rate and morphology.ResultsThe main outcomes of this study were that the sex ratio at birth of single blastocyst frozen thawed embryo transfer babies was 56.67% which was higher than the normal level (51.22%). The sex ratio of mothers older than 40 years was significantly lower than that of mothers younger than 40 years (0.39vs1.3, P<0.05). Transplanting good quality blastocysts significantly increased the proportion of boys (1.35vs0.94, P<0.05). Transplanting grade 5 and 6 blastocysts significantly increased the proportion of male babies born compared with grade 3 and 4 blastocysts (1.91vs1.28, P<0.05). There were no significant differences in the sex ratio with respect to the inner cell mass (ICM) score. In addition, the higher the trophectoderm (TE) score, the higher the sex ratio (2.79vs1.18vs0.91, P<0.05).ConclusionsOur study indicated that blastocyst quality, especially trophoblastic cell score, had a significant effect on sex ratio.

The Preeclamptic Environment Promotes the Activation of Transcription Factor Kappa B by P53/RSK1 Complex in a HTR8/SVneo Trophoblastic Cell Line

Preeclampsia is a pregnancy disorder associated with shallow placentation, forcing placental cells to live in hypoxic conditions. This activates the transcription factor kappa B (NFκB) in maternal and placental cells. Although the role of NFκB in preeclampsia is well documented, its mechanism of activation in trophoblastic cells has been never studied. This study investigates the mechanism of NFκB activation in a first trimester trophoblastic cell line (HTR8/SVneo) stimulated by a medium containing serum from preeclamptic (PE) or normotensive (C) women in hypoxic (2% O2) or normoxic (8% O2) conditions. The results indicate that in HTR8/SVneo cells, the most widely studied NFκB pathways, i.e., canonical, non-canonical and atypical, are downregulated in environment PE 2% O2 in comparison to C 8% O2. Therefore, other pathways may be responsible for NFκB activation. One such pathway depends on the activation of NFκB by the p53/RSK1 complex through its phosphorylation at Serine 536 (pNFκB Ser536). The data generated by our study show that inhibition of the p53/RSK1 pathway by p53-targeted siRNA results in a depletion of pNFκB Ser536 in the nucleus, but only in cells incubated with PE serum at 2% O2. Thus, the p53/RSK1 complex might play a critical role in the activation of NFκB in trophoblastic cells and preeclamptic placentas.

Low Abundance Fusobacterium Nucleatum Supports Early Pregnancy Development – An In Vitro Study

Pregnancy success depends greatly on a balanced immune homeostasis. The detection of bacterial components in the upper reproductive tract in non-pregnant and pregnant women raised questions on its possible beneficial role in reproductive health. The local conditions that allow the presence of bacteria to harmonize with the establishment of pregnancy are still unknown. Among the described bacterial species in endometrial and placental samples, Fusobacterium nucleatum was found. It has been observed that F. nucleatum can induce tumorigenesis in colon carcinoma, a process that shares several features with embryo implantation. We propose that low concentrations of F. nucleatum may improve trophoblast function without exerting destructive responses. Inactivated F. nucleatum and E. coli were incubated with the trophoblastic cell lines HTR8/SVneo, BeWo, and JEG-3. Viability, proliferation, migratory capacity, invasiveness and the secretion of chemokines, other cytokines and matrix metalloproteinases were assessed. The presence of F. nucleatum significantly induced HTR8/SVneo invasion, accompanied by the secretion of soluble mediators (CXCL1, IL-6 and IL-8) and metalloproteinases (MMP-2 and MMP-9). However, as concentrations of F. nucleatum increased, these did not improve invasiveness, hindered migration, reduced cell viability and induced alterations in the cell cycle. Part of the F. nucleatum effects on cytokine release were reverted with the addition of a TLR4 blocking antibody. Other effects correlated with the level of expression of E-cadherin on the different cell lines tested. Low amounts of F. nucleatum promote invasion of HTR8/SVneo cells and induce the secretion of important mediators for pregnancy establishment. Some effects were independent of LPS and correlated with the expression of E-cadherin on trophoblasts.