Placenta In Early Pregnancy Research Articles

Suppression of GATA3 promotes epithelial-mesenchymal transition and simultaneous cellular senescence in human extravillous trophoblasts

The regulatory mechanism of the transcription factor GATA3 in the differentiation and maturation process of extravillous trophoblasts (EVT) in early pregnancy placenta, as well as its relevance to the occurrence of pregnancy disorders, remains poorly understood. This study leveraged single-cell RNA sequencing data from placental organoid models and placental tissue to explore the dynamic changes in GATA3 expression during EVT maturation. The expression pattern exhibited an initial upregulation followed by subsequent downregulation, with aberrant GATA3 localization observed in cases of recurrent miscarriage (RM). By identifying global targets regulated by GATA3 in primary placental EVT cells, JEG3, and HTR8/SVneo cell lines, this study offered insights into its regulatory mechanisms across different EVT cell models. Shared regulatory targets among these cell types and activation of trophoblast cell marker genes emphasized the importance of GATA3 in EVT differentiation and maturation. Knockdown of GATA3 in JEG3 cells led to repression of GATA3-induced epithelial-mesenchymal transition (EMT), as evidenced by changes in marker gene expression levels and enhanced migration ability. Additionally, interference with GATA3 accelerated cellular senescence, as indicated by reduced proliferation rates and increased activity levels for senescence-associated β-galactosidase enzyme, along with elevated expression levels for senescence-associated genes. This study provides comprehensive insights into the dual role of GATA3 in regulating EMT and cellular senescence during EVT differentiation, shedding light on the dynamic changes in GATA3 expression in normal and pathological placental conditions.

Velocity-selective arterial spin labeling perfusion measurements in 2nd trimester human placenta with varying BMI

IntroductionProper placental development is crucial to fetal health but is challenging to functionally assess non-invasively and is thus poorly characterized in populations. Body mass index (BMI) has been linked with adverse outcomes, but the causative mechanism is uncertain. Velocity-selective arterial spin labeling (VS-ASL) MRI provides a method to non-invasively measure placental perfusion with robustness to confounding transit time delays. In this study, we report on the measurement of perfusion in the human placenta in early pregnancy using velocity-selective arterial spin labeling (VS-ASL) MRI, comparing non-obese and obese participants. MethodsParticipants (N = 97) undergoing routine prenatal care were recruited and imaged with structural and VS-ASL perfusion MRI at 15 and 21 weeks gestation. Resulting perfusion images were analyzed with respect to obesity based on BMI, gestational age, and the presence of adverse outcomes. ResultsAt 15 weeks gestation BMI was not associated with placental perfusion or perfusion heterogeneity. However, at 21 weeks gestation BMI was associated with higher placental perfusion (p < 0.01) and a decrease in perfusion heterogeneity (p < 0.05). In alignment with past studies, perfusion values were also higher at 21 weeks compared to 15 weeks gestation. In a small cohort of participants with adverse outcomes, at 21 weeks lower perfusion was observed compared to participants with uncomplicated pregnancies. DiscussionThese results suggest low placental perfusion in the early second trimester may not be the culpable factor driving associations of obesity with adverse outcomes.

Spatial Localization of Eubacterial 16S rRNA in Early Pregnancy Placenta and Decidua.

Bacteria derived from the maternal circulation have been suggested to seed the human placenta during pregnancy leading to development of an intrinsic placental microbiome; however, other data indicates these bacteria are artifactual contaminants. Limited research on the localization of bacteria in human placental tissue is available, which may help differentiate resident placental bacteria from contaminants. This study spatially localizes bacteria in situ in normal late first to early second trimester human placenta by 16S rRNA chromogenic in situ hybridization and demonstrates patterns consistent with both contaminants and intraparenchymal signals. These results suggest that placental microbiome studies may benefit from spatial strategies that can exclude surface contamination.

Comparison of HCG Levels in Reagents Temperature 25°C and 8°C Using ELISA Method

HCG hormone is a hormone produced by the placenta in early pregnancy with the aim of maintaining pregnancy. The Elisa method is a method for examining HCG levels which has the principle of specific antigen and antibody reactions. The work stage in the laboratory that has the biggest error is in the pre-analytic stage reaching 60%, one of which is the reagent temperature. The ELISA method recommends the reagent temperature to be stored at 2-8°C and when used at 25°C. Examination reagents Elisa method uses reagents that use enzymes in the examination. Enzymes are very sensitive to temperature. This study aims to determine the ratio of HCG levels at reagent temperatures of 25°C &amp; 8° using the ELISA method. This type of research is analytic supported by experiments and literature studies. Serum samples of 6 pregnant women were taken from the Kedungmundu Public Health Center using a limited population count formula. The examination was carried out by comparing the reagents at a temperature of 25°C and 8°C using 6 serums of pregnant women and in duplicate. Data analysis only used primary data and statistical tests were carried out using normality, homogeneity and T-Test tests. The average result at a temperature of 25°C was 380.851 and the average temperature of 8°C was 321.99 and the T-test statistic test showed p value = 0.469 and 0.470. Based on the results of the study, it can be concluded that there is an insignificant difference in the concentration of HCG by comparing the reagents at 25°C and 8°C.

Spatial multiomics map of trophoblast development in early pregnancy

The relationship between the human placenta—the extraembryonic organ made by the fetus, and the decidua—the mucosal layer of the uterus, is essential to nurture and protect the fetus during pregnancy. Extravillous trophoblast cells (EVTs) derived from placental villi infiltrate the decidua, transforming the maternal arteries into high-conductance vessels1. Defects in trophoblast invasion and arterial transformation established during early pregnancy underlie common pregnancy disorders such as pre-eclampsia2. Here we have generated a spatially resolved multiomics single-cell atlas of the entire human maternal–fetal interface including the myometrium, which enables us to resolve the full trajectory of trophoblast differentiation. We have used this cellular map to infer the possible transcription factors mediating EVT invasion and show that they are preserved in in vitro models of EVT differentiation from primary trophoblast organoids3,4 and trophoblast stem cells5. We define the transcriptomes of the final cell states of trophoblast invasion: placental bed giant cells (fused multinucleated EVTs) and endovascular EVTs (which form plugs inside the maternal arteries). We predict the cell–cell communication events contributing to trophoblast invasion and placental bed giant cell formation, and model the dual role of interstitial EVTs and endovascular EVTs in mediating arterial transformation during early pregnancy. Together, our data provide a comprehensive analysis of postimplantation trophoblast differentiation that can be used to inform the design of experimental models of the human placenta in early pregnancy.

Elevated PGT promotes proliferation and inhibits cell apoptosis in preeclampsia by Erk signaling pathway

Prostaglandins participate in maternal recognition of pregnancy, implantation and maintenance of gestation. Prostaglandin transporter (PGT), as a candidate molecule of prostaglandin carriers, might be involved in the pathogenesis of preeclampsia. In preeclampsia (PE) patients’ placental tissue, we identified PGT by RNA sequencing, measured its expression pattern by quantitative real-time PCR and Western blot. PGT was found to be upregulated in preeclamptic placental tissue. The expression pattern of PGT in PE was double confirmed by eight Gene Expression Omnibus (GEO) databases. In abortion tissues at 6–8 weeks, we then observed the cellular location of PGT by Immunofluorescence technique (IF) and found PGT located in trophoblast cell of the placenta of early pregnancy. In vitro studies revealed that forced expression of PGT in HTR8/Sveno cell inhibited its apoptosis, but promoted its proliferation by activating Erk signaling. In vivo study, we used reduced uterine perfusion pressure (RUPP) rat model and L-NAME-induced preeclampsia-like rats to study the possible role of PGT in preeclampsia. And PGT was found to be upregulated in both preeclampsia rat models by Immunohistochemical (IHC) staining. Newly identified PGT plays an important role in trophoblast proliferation via Erk signaling, providing new insights for understanding the pathogenesis of PE.

NINJ1 triggers extravillous trophoblast cell dysfunction through blocking the STAT3 signaling pathway.

Trophoblasts are the most important parts of the placenta in early pregnancy. Trophoblast cell dysfunction can induce embryo implantation insufficiency, thereby resulting in multiple diseases, including recurrent spontaneous abortion (RSA). A previous study indicates higher nerve injury-induced protein 1 (NINJ1) RNA levels in the villi tissues of RSA patients. This study aimed to investigate the effect of NINJ1 on trophoblast behaviors and pregnancy loss. Fresh villi tissues were obtained from with RSA patients and patients with artificial selective abortion for personal reasons, and NINJ1 expression in these tissues was detected. Extravillous trophoblast cell line HTR-8/SVneo was transfected with small-interfering RNA targeting NINJ1 or NINJ1 overexpression vector to perform loss-/gain-of-function experiments. Spontaneous abortion (SA) was induced by mating CBA/J females with DBA/2 males and the pregnant females were intraperitoneally injected with adenovirus vector carrying NINJ1 short hairpin RNA. NINJ1 mRNA and protein levels were higher in the villi tissues of RSA patients than those of artificial selective abortion patients. NINJ1 knockdown promoted trophoblast cell proliferation, migration and invasion but inhibited cell apoptosis. Moreover, conditioned medium from NINJ1-depleted trophoblasts promoted the angiogenesis of human umbilical vein endothelial cells. NINJ1 knockdown also promoted activation of the signal transducer and activator of transcription 3 (STAT3) signaling pathway in trophoblasts, and STAT3 inhibitor reversed NINJ1 knockdown-induced effects on trophoblast behaviors. Furthermore, pregnancy loss was attenuated by NINJ1 inhibition. NINJ1 contributes to the development of SA and triggers trophoblast cell dysfunction through inhibiting the STAT3 pathway.

Non-Coding RNAs and Prediction of Preeclampsia in the First Trimester of Pregnancy.

Preeclampsia (PE) is a major cause of maternal and perinatal morbidity and mortality. The only fundamental treatment for PE is the termination of pregnancy. Therefore, not only severe maternal complications but also perinatal complications due to immaturity of the infant associated with early delivery are serious issues. The treatment and prevention of preterm onset preeclampsia (POPE) are challenging. In 2017, the ASPRE trial showed that a low oral dose of aspirin administered to POPE high-risk women in early pregnancy reduced POPE by 62%. A prediction algorithm at 11–13 weeks of gestation identifies POPE with 75% sensitivity when the false positive rate is set at 10%. New biomarkers to increase the accuracy of the prediction model for POPE high-risk women in early pregnancy are needed. In this review, we focused on non-coding RNAs (ncRNAs) as potential biomarkers for the prediction of POPE. Highly expressed ncRNAs in the placenta in early pregnancy may play crucial roles in placentation. Furthermore, placenta-specific ncRNAs have been detected in maternal blood. In this review, we summarized ncRNAs that were highly expressed in the primary human placenta in early pregnancy. We also presented highly expressed ncRNAs in the placenta that were associated with or predictive of the development of PE in an expression analysis of maternal blood during the first trimester of pregnancy. These previous studies showed that the chromosome 19 microRNA (miRNA) -derived miRNAs (e.g., miR-517-5p, miR-518b, and miR-520h), the hypoxia-inducible miRNA (miR-210), and long non-coding RNA H19, were not only highly expressed in the early placenta but were also significantly up-regulated in the blood at early gestation in pregnant women who later developed PE. These maternal circulating ncRNAs in early pregnancy are expected to be possible biomarkers for POPE.

MiR-187 regulates the proliferation, migration and invasion of human trophoblast cells by repressing BCL6-mediated activation of PI3K/AKT signaling.

Recurrent miscarriage (RM), refers to two or more consecutive spontaneous miscarriage in a pregnant woman. RM is caused by many factors, and microRNAs play an important role in the development and pathology of RM. In the present study, we investigated the function of miR-187 in the pathogenesis of RM and its effects on human trophoblast cells. The localization of miR-187 in the human placenta in early pregnancy was determined by in situ hybridization. QRT-PCR was used to detect the expression of miR-187 in villi of normal early pregnancy induced abortion group and recurrent spontaneous miscarriage group. Then, HTR8/SVneo cells were used to investigated the effect of miR-187 on BCL6 expression and biological activity of trophoblasts. We found that the expression of miR-187 in villi of RM group was higher than that of normal abortion group and miR-187 inhibited the proliferation, migration, and invasion of HTR8 cells. We also found that miR-187 promoted apoptosis, inhibited EMT, and inhibited the PI3K/AKT pathway in HTR8 cells. In addition, we also found that BCL6 is a direct target of miR-187 and is negatively regulated by miR-187. In addition, BCL6 reversed the inhibitory effects of miR-187 on HTR8/SVneo cells. These data demonstrate that miR-187-induced repression of PI3K/AKT signaling is mediated by BCL6 in HTR8 cells. MiR-187 inhibits the proliferation, migration, and invasion of trophoblasts through a mechanism that involves regulation of BCL6.

MicroRNA-210/ Long non-coding RNA MEG3 axis inhibits trophoblast cell migration and invasion by suppressing EMT process

IntroductionPre-eclampsia is characterized by insufficient spiral artery remodeling, and trophoblast dysfunction plays an important role in this process. Noncoding RNAs (microRNAs (miRNAs) and long noncoding RNAs (LncRNAs) are included) can regulate trophoblasts. MicroRNA-210 (miR-210) can decrease trophoblast cell migration and invasion and may act as a biomarker for preeclampsia. LncRNA maternally expressed gene 3 (MEG3) plays a positive role in pre-eclampsia, and MEG3 can be a downstream target of miR-210 in human umbilical vein endothelial cells (HUVEC). However, the effect of miR-210 on MEG3 and the mechanism of action of the miR-210/MEG3 axis in trophoblasts remain unclear. MethodThe localization of miR-210 and MEG3 in the human placenta in early pregnancy was determined by in situ hybridization. Then, HTR8/SVneo cells were used to investigated the effect of miR-210 on MEG3 expression and biological activity of trophoblasts in the migration and invasion assays. Gain- and loss-of-function experiments were performed to determine the mechanism of action of miR-210 and MEG3 in Epithelial-mesenchymal transition (EMT) of HTR8/SVneo cells. ResultsThe qRT-PCR and western blotting results demonstrated that the upregulation of miR-210 decreases MEG3, N-cadherin and Vimentin expression and increases E-cadherin expression to inhibit EMT in HTR-8/SVneo cells. Inhibition of the expression of miR-210 results in the opposite effects. Gain- and loss-of-function assay indicated that miR-210 can impair the EMT, migration, and invasion of HTR8/SVneo cells by regulating the expression of MEG3. DisscussionMiR-210 may be a negative regulator of trophoblast EMT that acts by suppressing MEG3 expression.

Scoring Model for Prediction of Placenta Accreta Spectrum (PAS) in Early Pregnancy

Aim: The objective of this prospective cohort study is to create a scoring model for prediction of placenta accreta in early pregnancy by two-dimensional ultrasound and color Doppler.Materials and Methods: In our study, we investigated 146 Patients at high risk of PAS using two dimensional and color Doppler ultrasound for findings suggestive of placenta accreta in early pregnancy. At time of delivery, diagnosis of placenta accreta was confirmed clinically.Results: Significant Doppler and 2D ultrasound findings for prediction for PAS in early pregnancy were; low implantation of the gestational sac, presence of placental lacunae and gestational sac or placenta overlapping uterine scar by 2D transvaginal ultrasound and Intraplacental dilated vessels by Doppler ultrasound. A scoring model for prediction of PAS in early pregnancy was created using these significant findings in addition to the number of previous CS deliveries. A cut-off point of 4.5 (out of 7.5 total score value) was used for prediction of cases at high risk of placenta accreta with sensitivity 77% and specificity 95%.Conclusion: A standardized risk assessment scoring model based on number of previous cesarean deliveries, 2D- ultrasound and Doppler findings can predict women at highest risk for morbidly adherent placenta in early pregnancy with relatively high specificity.

The Role of TNF-α and Anti-TNF-α Agents during Preconception, Pregnancy, and Breastfeeding.

Tumor necrosis factor-alpha (TNF-α) is a multifunctional Th1 cytokine and one of the most important inflammatory cytokines. In pregnancy, TNF-α influences hormone synthesis, placental architecture, and embryonic development. It was also shown that increased levels of TNF-α are associated with pregnancy loss and preeclampsia. Increased TNF-α levels in complicated pregnancy draw attention to trophoblast biology, especially migratory activity, syncytialisation, and endocrine function. Additionally, elevated TNF-α levels may affect the maternal-fetal relationship by altering the secretory profile of placental immunomodulatory factors, which in turn affects maternal immune cells. There is growing evidence that metabolic/pro-inflammatory cytokines can program early placental functions and growth in the first trimester of pregnancy. Furthermore, early pregnancy placenta has a direct impact on fetal development and maternal immune system diseases that release inflammatory (e.g., TNF-α) and immunomodulatory factors, such as chronic inflammatory rheumatic, gastroenterological, or dermatological diseases, and may result in an abnormal release of cytokines and chemokines in syncytiotrophoblasts. Pregnancy poses a challenge in the treatment of chronic disease in patients who plan to have children. The activity of the disease, the impact of pregnancy on the course of the disease, and the safety of pharmacotherapy, including anti-rheumatic agents, in pregnancy should be considered.

Використання біомагнієвого комплексу Магнокс Прегна у профілактиці і лікуванні ускладнень вагітності при дефіциті магнію

Recent studies indicate a widespread deficiency of vitamins and minerals among pregnant and lactating women around the world. Most of the women examined (up to 80%), regardless of age, place of residence and professional affiliation, have a combined deficiency of three or more vitamins. Even with the most balanced diet, a diet adequate to the pregnant woman’s energy consumption is not able to provide the growing needs of the body with the necessary amount of minerals and microelements, which contribute to a change in the size and functional activity of the mother’s organs and systems during pregnancy, normal formation of the placenta, and fetal development. Magnesium deficiency leads to a violation of the processes of placentation and the threat of termination of pregnancy in the first trimester. The success of pregnancy largely depends on adequate implantation, the transformation of the spiral arteries and placentation with the establishment of full blood flow in the mother – placenta – fetus system. Violation of the formation of the placenta in early pregnancy leads subsequently to the development of placental dysfunction. Such a “physiological” magnesium deficiency during pregnancy in the absence of adequate nutritional subsidies can increase the risk of early and late miscarriages, premature births due to increased contractile activity of the myometrium, preeclampsia, etc. Chronic magnesium deficiency, which often leads to serious pregnancy complications, must be compensated for by the ingestion of magnesium supplements. Moreover, due to the fact that a mainly long course of treatment is required, in addition to the effectiveness of the action of magnesium-containing drugs, the bioavailability of magnesium and the safety of use are extremely important. Magnox Pregna biomagnesium complex is a unique combination of magnesium and vitamins B1, B6 and E, the use of which helps to suppress contractile activity of smooth and striated muscles (including the uterus), prevent and eliminate leg cramps, reduce the risk of developing hypertension, and the occurrence of eclampsia and premature birth. Key words: magnesium deficiency, vitamins, trace elements, pregnancy, complications, Magnox Pregna biomagnesium complex.

Role of ultrasound in the diagnosis and management of gestational trophoblastic disease in Rwanda Rural health facilities- A case report

Introduction Gestational trophoblastic disease (GTD) is a rare kind of proliferative disorder of trophoblastic cells which develops from the placenta in early pregnancy and the complete hydatidiform mole (CHM) sub-type which limited to endometrium is most common. It has excellent prognosis if early appropriate diagnosis and management are done. A well performed ultrasound(US) play a primordial role in the diagnosis of maternal health disorders during routine prenatal care. This helps in avoiding complications and consequently aids in achieving the objectives of the Millennium Development Goals (MDGs) in Rwanda. Aim To understand the definition of Gestational trophoblastic disease(GTD); To recognize key diagnostic findings of complete molar pregnancy on ultrasound and appropriate management in maternal follow up and To review the differential diagnosis for ultrasound findings seen with GTD. Methods The case has been referred in ultrasound unit for scan from outpatient department and follow up US was ordered and done during management period while the patient lies on a bed while operator moves the 3,5 MHz lubricated transducer of frequency with gel on the abdomino- pelvic region with full urinary bladder and Mindray ultrasound Machine has been used. Results There was a large moderately hyperechogenic mass completely filling the endometrial cavity with numerous small cystic spaces of different size. Post 5 days under treatment, anechoic area within endometrial cavity and myometrium well outlined was discharged and in 3rd medical visit after 2 weeks, US were normal. Conclusion Ultrasound is the first modality to be used in all rural health facilities for diagnosis of suspected Gestational trophoblastic disease (GTD) for better results. It is available and free of radiation. Recommendation -The US is the baseline modalities to be used in 1st trimester in pregnancy women with GTD suspicion bleeding. -All rural health facilities should have empowered and capable trained Medical imaging practitioners for appropriate carrying out the ultrasonographic investigation for better care service delivery on GTD(Medical imaging practitioners Role extension project after formal training by Prof. Kawooya).

IGF signalling and endocytosis in the human villous placenta in early pregnancy as revealed by comparing quantum dot conjugates with a soluble ligand.

A complex combination of trafficking and signalling occurs at the surface of the placenta. The system delivers maternal nutrients to the fetus and facilitates gaseous exchange, whilst mediating signal transduction to support and stimulate the growth of the placenta itself. IGF-I is acknowledged as a maternally-derived ligand important in the regulation of placental growth. Here we show that quantum dots bearing IGF can stimulate IGF receptor (IGF1R) phosphorylation in the syncytio- (maternal-facing) and cyto- (fetal-facing) trophoblast bilayer that forms the outer boundary of the placenta, in a distribution similar to the one resulting from exposure to a soluble ligand. The conjugates are internalised by a clathrin-dependent pathway and delivered to a syncytioplasmic compartment that differs from conventional late endosomes and lysosomes. Two discrete downstream responses are evident in different cellular compartments: phosphorylation of P70S6K in the non-proliferative syncytiotrophoblast and of AKT in the cytotrophoblast. Co-conjugation of IGF-quantum dots with an RGD-containing ligand permits penetration beyond the syncytium, into the cytoplasm of the underlying cytotrophoblast. These data reveal the existence of a trans-syncytial pathway that allows maternal mitotic signals to penetrate to the inner progenitor cells, which must proliferate to support placental and consequently fetal growth.

The early pregnancy placenta foreshadows DNA methylation alterations of solid tumors

ABSTRACTThe placenta relies on phenotypes that are characteristic of cancer to successfully implant the embryo in the uterus during early pregnancy. Notably, it has to invade its host tissues, promote angiogenesis—while surviving hypoxia—, and escape the immune system. Similarities in DNA methylation patterns between the placenta and cancers suggest that common epigenetic mechanisms may be involved in regulating these behaviors. We show here that megabase-scale patterns of hypomethylation distinguish first from third trimester chorionic villi in the placenta, and that these patterns mirror those that distinguish many tumors from corresponding normal tissues. We confirmed these findings in villous cytotrophoblasts isolated from the placenta and identified a time window at the end of the first trimester, when these cells come into contact with maternal blood, as the likely time period for the methylome alterations. Furthermore, the large genomic regions affected by these patterns of hypomethylation encompass genes involved in pathways related to epithelial-mesenchymal transition, immune response, and inflammation. Analyses of expression profiles corresponding to genes in these hypomethylated regions in colon adenocarcinoma tumors point to networks of differentially expressed genes previously implicated in carcinogenesis and placentogenesis, where nuclear factor kappa B is a key hub. Taken together, our results suggest the existence of epigenetic switches involving large-scale changes of methylation in the placenta during pregnancy and in tumors during neoplastic transformation. The characterization of such epigenetic switches might lead to the identification of biomarkers and drug targets in oncology as well as in obstetrics and gynecology.

Up-and-down immunity of pregnancy in humans.

One part of the human placenta in early pregnancy is particularly important for local immunity: the decidua basalis, which is transformed endometrium located at the site of embryo implantation . This placental bed tissue contains both maternal uterine immune cells, including decidual natural killer (NK) cells, the dominant leukocyte population exhibiting a unique phenotype, and fetal extravillous trophoblast which comes into direct contact with maternal decidual cells . To establish a successful placental development and healthy pregnancy outcome, the maternal immune system must tolerate paternal antigens expressed by trophoblast cells yet remain efficient for clearing any local pathogen infection. This review deals mainly with decidual NK cells. A key element, among others, to achieve such dual functions is the direct interaction between activating and inhibitory receptors expressed by decidual NK cells and their specific ligands presented by trophoblast or other decidual cells. Depending whether maternal decidual cells and trophoblast are infected by viruses, the balance between activating and inhibitory receptor signals mediated by decidual NK cell–trophoblast cross-talk results in tolerance (healthy pregnancy) or specific killing (pathogen-infected cells).

СТАН СУДИННОГО ГОМЕОСТАЗУ У ЖІНОК З НЕПЛІДДЯМ ПІСЛЯ ВИКОРИСТАННЯ ДОПОМІЖНИХ РЕПРОДУКТИВНИХ ТЕХНОЛОГІЙ.

Мета – встановити роль порушень судинної регуляції у виникненні перинатальних ускладнень у вагітних високого ризику.Методи дослідження. Обстежено 50 вагітних жінок після допоміжних репродуктивних технологій із непліддям гормонального генезу (основна група), 50 вагітних жінок після допоміжних репродуктивних технологій внаслідок трубно-перитонеального фактору непліддя (група порівняння) і 50 соматично здорових жінок із неіндукованою вагітністю (контрольна група) у терміні гестації 14–16 тижнів. Рівень гомоцистеїну визначено методом ферментативної циклічної реакції, за допомогою набору Diasyis (Німеччина) з використанням аналізатора Respons 920 (Німеччина) за методикою виробника. Вміст L"аргініну в крові визначено за допомогою фотометричного методу, в основу якого покладено реакцію L"нафтола з гіпобромідним реактивом. Для кількісного визначення плацентарного фактора росту людини (PLGF) в зразках плазми крові використано імунохімічний метод з електрохемілюмінесцентною детекцією (ECLIA).Результати. Для визначення ролі порушень регуляції судинного гомеостазу у виникненні ускладнень вагітності в основній групі виділено 2 підгрупи: 1"ша – 9 жінок із суттєвими акушерськими й перинатальними порушеннями (викидні та завмерлі вагітності, прееклампсія тяжкого ступеня, декомпенсований дистрес плода), 2"га – 31 жінка без таких порушень. Встановлено, що вагітність після програм допоміжних репродуктивних технологій проходила з особливо тяжкими ускладненнями в тих жінок із непліддям гормонального генезу, в яких вже на початку вагітності формування і розвиток плаценти відбувались в умовах ендотеліальної дисфункції: негативної дії гомоцистеїну, порушень процесів ангіогенезу.Отже, ці фактори можуть бути ранніми прогностичними маркерами тяжких акушерських і перинатальних порушень при індукованій вагітності в жінок із непліддям різного генезу. Висновки. В умовах гомеостатичного дисбалансу в даної категорії жінок порушення ендотелію та факторів судинної регуляції лежать в основі патогенезу репродуктивних, акушерських і перинатальних ускладнень, що є теоретичною передумовою для розробки патогенетично обґрунтованого комплексу профілактики й лікування, спрямованого на корекцію виявлених порушень.

Vulnerability of primitive human placental trophoblast to Zika virus

Infection of pregnant women by Asian lineage strains of Zika virus (ZIKV) has been linked to brain abnormalities in their infants, yet it is uncertain when during pregnancy the human conceptus is most vulnerable to the virus. We have examined two models to study susceptibility of human placental trophoblast to ZIKV: cytotrophoblast and syncytiotrophoblast derived from placental villi at term and colonies of trophoblast differentiated from embryonic stem cells (ESC). The latter appear to be analogous to the primitive placenta formed during implantation. The cells from term placentas, which resist infection, do not express genes encoding most attachment factors implicated in ZIKV entry but do express many genes associated with antiviral defense. By contrast, the ESC-derived trophoblasts possess a wide range of attachment factors for ZIKV entry and lack components of a robust antiviral response system. These cells, particularly areas of syncytiotrophoblast within the colonies, quickly become infected, produce infectious virus and undergo lysis within 48 h after exposure to low titers (multiplicity of infection > 0.07) of an African lineage strain (MR766 Uganda: ZIKVU) considered to be benign with regards to effects on fetal development. Unexpectedly, lytic effects required significantly higher titers of the presumed more virulent FSS13025 Cambodia (ZIKVC). Our data suggest that the developing fetus might be most vulnerable to ZIKV early in the first trimester before a protective zone of mature villous trophoblast has been established. Additionally, MR766 is highly trophic toward primitive trophoblast, which may put the early conceptus of an infected mother at high risk for destruction.

36 The role of foxm1 in early placentation in normal pregnancy

Introduction The development of placenta plays a central role in determining the outcome of pregnancy. Extravillous trophoblast invasion is a fundamental component of human placentation. However, the transcriptional networks responsible for controlling the extent of trophoblast invasion are not well defined. Trophoblast cells are often compared to highly invasive carcinoma cells due to their capacity to proliferate and invade and to exhibit similar gene expression signatures. Thus, the transcriptional regulation of genes associated with cancer metastasis and trophoblast invasion may also be shared. The transcription factor forkhead box M1 (FOXM1), which is one of a few genes shown to be upregulated during early cancer development, possibly fits this scenario. FOXM1 has been shown to play a crucial role in the regulation of cell proliferation, cell cycle progression, angiogenesis, metastasis and DNA damage repair. Objective our objective in this study was to evaluate the role of FOXM1 in normal placental development. Method To evaluate the expression of FOXM1 throughout pregnancy Sprague Dawley dams were sacrificed at E14, E15, E18, E19 and E20 and protein and total RNA was isolated from one placenta per dam. Changes in placental protein expression were analyzed by western blot and mRNA expression was analyzed by qPCR. The influence of FOXM1 on trophoblast proliferation and migration was evaluated by Sulforhodamine B colorimetric (SRB) assay and wound healing assay, respectively, in JEG cell lines following silencing of FOXM1 via RNA interference under 21%, 8% and 3% oxygen conditions. Results The transcription regulator FOXM1 was expressed during early-mid gestation (E14) but its expression decreased as gestational age increased (E14 to E18 there was a decrease in 0.29 fold and E14 to E21 0.04 fold). Compared to trophoblast cells expressing control non-specific siRNA, FOXM1 knockdown resulted in a significant reduction of migration at 21%, 8% and 3% O 2 , whereas silencing of FOXM1 slows down the proliferation of JEG-3 cells at 21% O 2 but it does not alter its proliferative capacity at 8% or 3% O 2 . Conclusions Our data is suggestive that FOXM1 modulates trophoblast migration but not proliferative potential at 8% and 3% O 2 , which coincides with oxygen tension in the placenta in early pregnancy and the results of the rat placenta indicate that FOXM1 may play a role in early placentation events. However, more studies are required to fully understand the exact role of FOXM1 in placenta development.