Regulation Of Trophoblast Invasion Research Articles

Feto-maternal interactions and immunological tolerance of the mother to her semiallogeneic fetus

Pregnancy is a cooperative interaction between the mother and her fetus, allowing survival and normal growth of the fetus. Successful pregnancy remains a fascinating phenomenon as it resists the immunological rules of rejection. Immunological recognition of the fetus is vital for maintenance of gestation. The maternal immune system undergoes changes that lead to tolerance of the fetus. Inadequate recognition of fetal antigens may cause abortion. In fact, fetal cells express paternal alloantigens that are not recognized as foreign by the mother. A special balance between lymphocytes is present at the feto-maternal interface to control the immune response. In addition, placental trophoblasts act as a physical barrier and exert an immunoregulatory function. Trophoblast cells regulate the expression of human leucocyte antigens. Dysfunction of these cells leads to morphological and functional alterations of the feto-maternal barrier as well as to recurrent spontaneous abortions. Uterine natural killer cells are appropriate residents of the materno-fetal interface to support the adaptation of the blood vessels of the pregnant uterus and regulate trophoblast invasion into the decidua and myometrium. Cytokines are involved at the feto-placental unit by adapting normal T-cell trafficking and modulating the inflammatory process. This study discusses the complex immunological aspects of immune tolerance and the balance of immunity in pregnancy in terms of the role of the human leucocyte antigen, placental trophoblasts, maternal immunosuppression, immune cells, cytokines and immunoregulatory molecules at the feto-maternal interface.

The role of progesterone in implantation and trophoblast invasion

With its genomic and non-genomic actions, progesterone plays a role in preparing the endometrium for implantation and also in regulating trophoblast invasion and migration. The genomic actions of progesterone are mediated by the classical nuclear progesterone receptors, PR-A and PR-B. In addition to their genomic actions, nuclear progesterone receptors may also trigger rapid cytoplasmic signalling events. Membrane-bound progesterone receptors have been implicated in the rapid non-genomic actions of progesterone. Both genomic and extra-nuclear actions of progesterone are crucial for adequate decidualisation and implantation. Progesterone plays a role in establishing uterine receptivity by blocking the proliferative effect of oestrogen, by inducing genes that allow the endometrium to permit embryo attachment, and also acts as a negative regulator of trophoblast invasion by controlling matrix metalloproteinase (MMP) activity.

Intermedin/Adrenomedullin 2 Is Associated With Implantation and Placentation via Trophoblast Invasion in Human Pregnancy

Intermedin (IMD) is a novel peptide expressed in trophoblast cells in human placenta and enhances the invasion, migration, and human leukocyte antigen class I, G (HLA-G) expression in first-trimester HTR-8SV/neo cells. We recently reported that infusion of IMD antagonist in pregnant rats is detrimental to pregnancy outcome, resulting in impaired fetoplacental growth and deformed placental vasculature. This study was undertaken to assess expression of IMD and its involvement in human implantation and early placentation and assess whether its expression is altered in spontaneous abortion. We demonstrate for the first time that IMD is present in day 5 embryonic secretome; villous and decidual expression of IMD is higher at 6-8 weeks after a decline as gestation advances toward the second trimester; first-trimester spontaneous abortion is associated with a lower expression of IMD in serum, villi, and decidua; IMD stimulates the invasive capacity of first-trimester primary Extravillous cytotrophoblast cells; and IMD decreases elevated levels of tumor suppressor Kangia-1 in decidual explants from first-trimester spontaneous abortion. In conclusion, this study is the first to demonstrate a potential involvement of IMD in human embryo implantation and placental development via regulation of trophoblast invasion at the maternal-fetal interface and suggests a physiological role for this novel peptide in establishment of human pregnancy.

580: A pathway analysis of imprinted genes in developing placentae

genes in developing placentae Jennifer King, Kai Wang, Huaiyu Mi, Nicole Bender, Rachel Steward, Sue Ingles, Joseph Ouzounian, Melissa Wilson University of Southern California, Department of Obstetrics and Gynecology, Los Angeles, CA, University of Southern California, Department of Psychiatry and The Behavioral Sciences, Los Angeles, CA, University of Southern California, Department of Preventive Medicine, Los Angeles, CA OBJECTIVE: Genomic imprinting has been demonstrated to be integral to fetal development and placentation. The majority of studies on imprinted gene expression in humans have involved term placentas, while placental imprinting is known to be a dynamic process, which evolves during human pregnancy. For these reasons, the distinct imprinted gene expression profile displayed in the early human placenta remains largely unknown. Our objective was to investigate the pattern of imprinted gene expression across first and early second trimester placental development. STUDY DESIGN: We sequenced complementary cDNA from placental villi obtained at 7, 8, 9, and 15 weeks gestation using massively parallel next-generation sequencing (RNA-seq). We used the PANTHER classification system (http://www.pantherdb.org/about.jsp) to evaluate imprinted gene expression at each gestational age in reference to the human genome. We performed a pathway analysis to determine important processes at each time point that differ from the human genome using the Wilcoxon Rank Sum test. We did not use a Bonferroni correction due to the exploratory nature of the analysis. We report the top 15 pathways for each gestational age. RESULTS: Table includes top 15 pathways for each gestational age, none of which are statistically significant after Bonferroni correction. CONCLUSION: Similarities between global gene expression pathways and imprinted gene pathways speak to the importance of imprinted genes in the developing placenta. Imprinted genes have variable expression in early placental development. The top pathway for gestational ages of 8 and 9 weeks appear remarkably similar. At 15 weeks, the molecular pathways of the imprinted genes are notably different compared to earlier gestational age. These observations may reflect changes in imprinted gene expression that occur with changes in oxygen pressure or may be a result of small sample size. Larger studies are required to differentiate. 581 A pathway analysis of the developing placentome Jennifer King, Kai Wang, Huaiyu Mi, Ian Tilley, Rachel Steward, Sue Ingles, Joseph Ouzounian, Melissa Wilson University of Southern California, Department of Obstetrics and Gynecology, Los Angeles, CA, University of Southern California, Department of Psychiatry and The Behavioral Sciences, Los Angeles, CA, University of Southern California, Department of Preventive Medicine, Los Angeles, CA OBJECTIVE: The creation of a suitable intrauterine environment for fetal development is dependent on the proper functioning of placental chorionic villi. Chorionic villi require well-coordinated expression of several transcription factors, cytokines and their receptors, growth factors, and cell cycle regulators. Disruption of the expression of these regulatory factors may lead to placental growth and development disorders. However, gene expression levels across gestatational age have not been fully delineated. Our objective was to investigate the pattern of global gene expression across first and early second trimester placental development. STUDY DESIGN: In the present investigation, we sequenced complementary cDNA from placenta villi obtained at 7, 8, 9, and 15 weeks gestation, using massively parallel next-generation sequencing (RNAseq). We used the PANTHER classification system (http://www.pantherdb.org/about.jsp) to evaluate global gene expression at each gestational age in reference to the human genome. We performed a pathway analysis to determine important processes at each time point that differ from the human genome using the Wilcoxon Rank Sum test with a Bonferroni correction for multiple comparisons. RESULTS: See Table. CONCLUSION: Pathways involved in protein degradation, and regulation of trophoblast invasion are important at all four gestational ages. However, the relative importance of different pathways diverges after that point among the four gestational ages. The possible clinical significance of the ubiquitin proteasome pathway may be underappreciated to date. Additional analyses with a larger sample size are required to understand how these pathways may play a role in predisposing to diseases of placental etiology, such as preeclampsia or intrauterine growth restriction. Pathway description, ( )over or ( )under represented vs human genome (reference) Poster Session IV Academic Issues, Antepartum Fetal, Clinical Ob, Fetus, Genetics, Hypertension, Med-Surg-Diseases, Operative Ob, U/S www.AJOG.org

Who Let the Dogs In: A Canine Trophoblast Invasion Model for Pre‐eclampsia

Pre-eclampsia affects 2-8% of pregnant women worldwide and is the third leading cause of maternal mortality in the United States, accounting for 20% of maternal deaths, for which the only known cure is delivery of the placenta. It is known that pre-eclampsia results from defects within the trophoblast invasion of the endometrium and myometrium. At a morphological level within the pre-eclamptic human placenta, trophoblast invasion is shallow, and this results in hypoperfusion, which is a life-threatening condition for both the mother and the foetus. Pre-eclampsia has been intensively investigated for over 50 years, and yet the causes are largely unknown. Despite a large body of data, it is still unknown exactly which mechanisms regulate trophoblast invasion. An effective animal model may be crucial to understanding the underlying causes of pre-eclampsia. A canine model is a proposed improvement on the current efforts to investigate disorders of shallow trophoblast invasion throughout gestation and to improve understanding of the factors that regulate trophoblast invasion. The objectives of this research were to elucidate and compare cellular and molecular similarities between normal canine trophoblasts with those from recently published reports on pre-eclampsia in women.

Gene Expression in Pre‐Term, Pre‐Labour and Parturient Canine Placenta

Matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), vascular endothelial growth factor (VEGF)-A, VEGF-A receptor (Flt-1) and KiSS-1 receptor (KiSS-1R) all play a role in trophoblast invasion in a number of mammalian species. However, mRNA expression of factors regulating trophoblast invasion has not been studied in dogs. Abnormal expression of these factors at the end of canine gestation may contribute to placental retention and/or subinvolution of placental sites. Therefore, we sought to determine the relative mRNA expression of these factors in canine chorioallantois tissue at 61 ± 1 day past the LH surge (pre-term; n = 4), following elective c-section at 64 ± 1 day past the LH surge prior to first-stage labour (pre-labour; n = 4) and following natural delivery (parturient; n = 3). Total RNA was isolated, real-time RT-PCR was performed, and relative expression was calculated using the relative quantitation (2-ΔΔCT) method. MMP-9 mRNA expression was significantly higher in pre-term samples compared to pre-labour and parturient samples. The results showed no significant difference between MMP-2, TIMP-2, VEGF-A and Flt-1 mRNA expression among the three groups. KiSS-1R mRNA was not expressed in any tissues studied. Gene expression of MMP-9 may be related to the onset of labour, whereas MMP-2, VEGF-A, Flt-1, TIMP-2 and KiSS-1R mRNA do not appear to play a role at the end of gestation in the dog.

High-Temperature Requirement Protein A4 (HtrA4) Suppresses the Fusogenic Activity of Syncytin-1 and Promotes Trophoblast Invasion

Cell-cell fusion and cell invasion are essential for placental development. Human cytotrophoblasts in the chorionic villi may undergo cell-cell fusion to form syncytiotrophoblasts to facilitate nutrient-gas exchange or differentiate into extravillous trophoblasts (EVTs) to facilitate maternal-fetal circulation. The placental transcription factor glial cells missing 1 (GCM1) regulates syncytin-1 and -2 expression to mediate trophoblast fusion. Interestingly, GCM1 and syncytin-1 are also expressed in EVTs with unknown physiological functions. In this study, we performed chromatin immunoprecipitation-on-chip (ChIP-chip) analysis and identified the gene for high-temperature requirement protein A4 (HtrA4) as a GCM1 target gene, which encodes a serine protease facilitating cleavage of fibronectin and invasion of placental cells. Importantly, HtrA4 is immunolocalized in EVTs at the maternal-fetal interface, and its expression is decreased by hypoxia and in preeclampsia, a pregnancy complication associated with placental hypoxia and shallow trophoblast invasion. We further demonstrate that HtrA4 interacts with syncytin-1 and suppresses cell-cell fusion. Therefore, HtrA4 may be crucial for EVT differentiation by playing a dual role in prevention of cell-cell fusion of EVTs and promotion of their invasion into the uterus. Our study reveals a novel function of GCM1 and HtrA4 in regulation of trophoblast invasion and that abnormal HrtA4 expression may contribute to shallow trophoblast invasion in preeclampsia.

Altered placental expression of kisspeptin and its receptor in pre-eclampsia

Kisspeptin, originally identified as metastatin, important in preventing cancer metastasis, has more recently been shown to be important in pregnancy. Roles indicated for kisspeptin in pregnancy include regulating trophoblast invasion and migration during placentation. The pregnancy-specific disorder pre-eclampsia (PE) is now accepted to begin with inadequate trophoblast invasion and the current study therefore sets out to characterise placental expression of both kisspeptin (KISS1) and its receptor (KISS1R) throughout pregnancy and in PE. Placental tissue was obtained from women undergoing elective surgical termination of early pregnancy (n=10) and from women following Caesarean section at term in normal pregnancy (n=10) and with PE (n=10). Immunohistochemistry of paraffin embedded sections and western immunoblotting were performed to assess protein localisation and expression. Quantitative real-time PCR was carried out to evaluate mRNA expression of both KISS1 and KISS1R. Protein and mRNA expression was found to mirror each other with KISS1 expression found to be reduced in PE compared with that in normal term pregnancy. Interestingly, KISS1R expression at both the mRNA and protein levels was found to be increased in PE compared with that in normal term pregnancy. The current findings of increased KISS1R expression may represent a mechanism by which functional activity of KISS1 is higher in PE than in normal pregnancy. Higher levels of activity of KISS1R may be involved in inhibition of trophoblast invasion and angiogenesis, which are associated with PE.

Gonadotropin-releasing hormone/gonadotropin-releasing hormone receptor signaling in the placenta

This review summarizes our current understanding of the role of gonadotropin-releasing hormone (GnRH)/GnRH receptor (GnRHR) signaling at the maternal-fetal interface. Several isoforms of GnRH and GnRHR are described. The hypothalamic decapeptide, GnRH-I, binds to the anterior pituitary and induces the synthesis and secretion of luteinizing hormone and follicle-stimulating hormone. It is also found in extrahypothalamic sites. A second isoform, GnRH-II, acts both in the hypothalamus and other organ systems, including placenta, breast, endometrium, and ovary. Although several putative isoforms of GnRHR have been identified, it is clear that, in humans, both GnRH-I and GnRH-II signal through a single receptor, GnRHR-I. GnRH-I, GnRH-II, and GnRHR-I mRNA and protein have been identified in placenta and regulate the β-subunit of human chorionic gonadotropin production, which is essential for the maintenance of early pregnancy. They may also play a role in the autocrine/paracrine regulation of trophoblast invasion through extracellular matrix remodeling. GnRH-I and GnRH-II have multiple extrapituitary roles. In placenta, they bind to GnRHR-I to stimulate the production of β-subunit of human chorionic gonadotropin. They may also play a role in trophoblast invasion. A better understanding of the molecular mechanisms involved in GnRH/GnRHR signaling at the maternal-fetal interface may identify novel roles for GnRH agonists/antagonists in the prevention or treatment of hormonally mediated diseases.

The pivotal role of protein inhibitors of activated STATs (PIAS) in regulating trophoblastic functions

Background: Signal Transducer and Activator of Transcription 3 (STAT3) is an intracellular signaling molecule which, in trophoblastic cells, can be phosphorylated at its tyr705 (p-STAT3 tyr705) residue by Leukemia Inhibitory Factor (LIF) receptor binding. This phosphorylation correlates with invasiveness. Protein inhibitors of activated STATs (PIAS) antagonize STAT function by interfering with STAT DNA-binding and are, therefore, expected to regulate trophoblast invasion.

Immunomodulator expression in trophoblasts from the feline immunodeficiency virus (FIV)-infected cat

BackgroundFIV infection frequently compromises pregnancy under experimental conditions and is accompanied by aberrant expression of some placental cytokines. Trophoblasts produce numerous immunomodulators that play a role in placental development and pregnancy maintenance. We hypothesized that FIV infection may cause dysregulation of trophoblast immunomodulator expression, and aberrant expression of these molecules may potentiate inflammation and compromise pregnancy. The purpose of this project was to evaluate the expression of representative pro-(TNF-α, IFN-γ, IL-1β, IL-2, IL-6, IL-12p35, IL-12p40, IL-18, and GM-CSF) and anti-inflammatory cytokines (IL-4, IL-5, and IL-10); CD134, a secondary co-stimulatory molecule expressed on activated T cells (FIV primary receptor); the chemokine receptor CXCR4 (FIV co-receptor); SDF-1α, the chemokine ligand to CXCR4; and FIV gag in trophoblasts from early-and late-term pregnancy.MethodsWe used an anti-cytokeratin antibody in immunohistochemistry to identify trophoblasts selectively, collected these cells using laser capture microdissection, and extracted total RNA from the captured cell populations. Real time, reverse transcription-PCR was used to quantify gene expression.ResultsWe detected IL-4, IL-5, IL-6, IL-1β, IL-12p35, IL-12p40, and CXCR4 in trophoblasts from early-and late-term pregnancy. Expression of cytokines increased from early to late pregnancy in normal tissues. A clear, pro-inflammatory microenvironment was not evident in trophoblasts from FIV-infected queens at either stage of pregnancy. Reproductive failure was accompanied by down-regulation of both pro-and anti-inflammatory cytokines. CD134 was not detected in trophoblasts, and FIV gag was detected in only one of ten trophoblast specimens collected from FIV-infected queens.ConclusionFeline trophoblasts express an array of pro-and anti-inflammatory immunomodulators whose expression increases from early to late pregnancy in normal tissues. Non-viable pregnancies were associated with decreased expression of immunomodulators which regulate trophoblast invasion in other species. The detection of FIV RNA in trophoblasts was rare, suggesting that the high rate of reproductive failure in FIV-infected queens was not a direct result of viral replication in trophoblasts. The influence of placental immune cells on trophoblast function and pregnancy maintenance in the FIV-infected cat requires additional study.

Regulation of invasion behavior of ERK-mediated trophoblast by focal adhesion kinase

To investigate the role of focal adhesion kinase (FAK) in extracellular signal-regulated kinase (ERK) signaling pathway mediated invadsion of trophoblasts. We established a human extravillous cytotrophoblasts in vitro invasion model. Different concentrations of herbimycin A(FAK inhibitor)and PD98059 (ERK inhibitor) were given to observe the influence on the growth of trophoblast cells, FAK, ERK phosphorylation, and trophoblast invasion abilities. The expression of phosphorylated FAK in the extravillous cytotrophoblasts (EVCT) was inhibited by herbimycin A in a concentration-dependent manner and expression of phosphorylated ERK1/2 was also partially reduced. PD98059 had no effect on the expression of phosphorylated FAK. Herbimycin A and PD98059 suppressed the in vitro invasion of EVCT to various degrees. ERK signaling pathway may be the common pathway for many invasive signals,and play a key role in the regulation of trophoblast invasion.

Progesterone-induced blocking factor (PIBF) and trophoblast invasiveness

Controlled trophoblast invasion is a key process during human placentation and a prerequisite for successful pregnancy. Progesterone is one of the factors to regulate trophoblast invasiveness. Progesterone-induced blocking factor (PIBF) is a progesterone-induced molecule expressed by the trophoblast, and also by tumors. The distribution of PIBF within the first-trimester decidua coincides with sites of trophoblast invasion. Another molecule that has been implicated in the control of trophoblast invasiveness is placental leptin. Leptin inhibits the secretion of progesterone by cytotrophoblast. The aim of this work was to investigate the possible interaction of PIBF and leptins in regulating trophoblast invasion. Paraffin-embedded sections from normal first-trimester placentae, partial moles, complete moles, and choriocarcinomas were reacted with PIBF, leptin, and leptin receptor specific antibodies. PIBF-deficient trophoblast cells were generated using siRNA and leptin receptor was detected on Western blot analysis. The lysates of PIBF-treated cells were used for detecting leptin expression in a protein array. PIBF was expressed in both normal first-trimester villous trophoblast and in partial mole. Compared with this, PIBF expression was markedly decreased in complete mole and absent in choriocarcinoma. Neither leptinR nor leptin were detected in partial mole, whereas both of these molecules were present in complete mole and choriocarcinoma. Leptin receptor expression was upregulated in PIBF-deficient cells, while leptin expression was decreased in PIBF-treated cells. These data suggest that PIBF affects the expression of leptin and its receptor, and that PIBF expression is inversely related to trophoblast invasiveness.

Possible Roles for Folic Acid in the Regulation of Trophoblast Invasion and Placental Development in Normal Early Human Pregnancy1

In addition to its role in the prevention of neural tube defects, folic acid has many other physiological functions, including cell proliferation, DNA replication, and antioxidant protection. The aim of this study was to determine the role that folic acid has in regulating placental trophoblast development. Placental explants from placentae at gestational age 7 wk (n = 3) were cultured in folic acid at concentrations of 10(-6) M, 10(-8) M, and 10(-10) M. Extravillous trophoblast (EVT) invasion was assessed following 6-day culture, and explants were used for immunohistochemical evaluation of proliferation (MKI67) and apoptosis (active caspase 3). In addition, an array was performed on cell culture supernatants to examine a range of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs). Folic acid increased the invasion of EVT cells in this explant model by between 83% and 19% (P = 0.005), and this was associated with increased MKI67 positivity and decreased active caspase 3 positivity; this effect was concentration dependent and showed a biphasic response. In addition, culture in folic acid increased vascular density, as determined by anti-CD31 immunostaining (P = 0.05). The increase in EVT invasion correlated with increased placental explant secretion of MMP2 (P = 0.01), MMP3 (P = 0.01), and MMP9 (P = 0.02). This study demonstrates that folic acid is potentially important in a number of crucial early stages of placental development, including EVT invasion, angiogenesis, and secretion of MMPs, and highlights the need for further studies to address the benefit of longer-term folic acid supplementation throughout pregnancy to prevent pregnancy disorders associated with deficient placental development, including preeclampsia.

Decidual HtrA3 negatively regulates trophoblast invasion during human placentation

BACKGROUND Controlled trophoblast invasion into the maternal decidua (interstitial invasion) is important for placental development. Factors secreted by the maternal decidual cells and the extravillous trophoblast can influence trophoblast invasion and abnormalities in the invasion process may lead to pregnancy complications. Serine protease HtrA3 is highly expressed in the decidual cells in the late secretory phase of the menstrual cycle and throughout pregnancy, and in most trophoblast cell types, apart from the invading interstitial trophoblast during the first trimester. HtrA3 and its family members are down-regulated in a number of cancers and are proposed as tumour suppressors. The current study aimed to investigate whether HtrA3 is secreted by decidual cells, and whether inhibiting such secretion alters trophoblast invasion. METHODS AND RESULTS Human endometrial stromal cells (HESCs) were decidualized with estradiol, medroxyprogesterone acetate and cyclic adenosine monophosphate. Real-time RT-PCR, western blotting and immunocytochemistry confirmed that HtrA3 mRNA and protein expression increased during decidualization. HtrA3 was also detected in the conditioned media (CM) of the decidualized HESCs, confirming its secretion. For functional studies, a protease-inactive mutant form of HtrA3 which was previously confirmed to be a dominant-negative inhibitor was produced using wheat germ cell-free technology. CM from decidualized HESCs significantly suppressed invasion of trophoblast HTR-8 cells (P < 0.01), whereas inhibition of HtrA3 in this CM by exogenous HtrA3 mutant resulted in increased trophoblast HTR-8 cell invasion (P < 0.001). CONCLUSIONS These results strongly support the hypothesis that decidual HtrA3 negatively regulates trophoblast invasion.

Molecular regulation of trophoblast invasion

Molecular regulation of trophoblast invasion

P72 Decreased proportion of peripheral blood galectin-1-expressing T and natural killer cells in preeclampsia

The invasion and migration of trophoblast cells are essential steps of normal placentation and successful pregnancy. The process is well-regulated by many factors at the fetal–maternal surface. Inadequate invasion by trophoblast cells may lead to poor perfusion of the placenta or complications such as preeclampsia (PE). Accumulating evidence suggests that N-acetylglucosaminyltransferase V (MGAT5) is correlated with tumor invasion and metastasis. Our objective was to characterize MGAT5 expression and function during placental development.The expression of MGAT5 in humans in placental tissue from the first trimester was determined by immunohistochemistry. To investigate whether MGAT5 regulates trophoblast invasion and migration, we investigated invasion/migration of the HTR8/SVneo trophoblast cells and used human villous explants. Cell proliferation and apoptosis were measured by CCK-8 assay and flow cytometry, respectively. The activity of matrix metalloproteinase (MMP) 2/9, and the expression of tissue inhibitors of metalloproteinases (TIMPs) 1/2 were determined by gelatin zymography and Western blot, respectively.MGAT5 was specifically localized within the cytotrophoblast, the syncytiotrophoblast and the trophoblast columns of human placental villi, decidual cells and some extravillous cells in the maternal decidua. MGAT5 shRNA significantly enhanced the invasion and migration capability of HTR8/SVneo cells, and increased villous explant outgrowth but did not affect proliferation and apoptosis of the trophoblast. The enhanced effect of MGAT5 shRNA on trophoblast cell invasion was associated with increased gelatinolytic activity of MMP2/9 and decreased expression of TIMP1/2.Our data support a role for MGAT5 in the inhibition of human trophoblast cell invasion and migration during early pregnancy by direct or indirect regulation of MMP2/9 activity.

Bizarre expression of tissue inhibitor of metalloproteinase-3 in feto–maternal interface caused gestational hypertension in mice model

Department of Obstetrics and Gynaecololgy, Osaka University Graduate School of Medicine, Suita, Osaka, Japan Preeclampsia (PE) is believed to result from abnormal placentation in early pregnancy. Shallow endovascular cytotrophoblast invasion and endothelial cell dysfunction are two key features in the pathogenesis of PE. Matrix metalloproteinase-9 (MMP-9) is crucial for the invasive ability of trophoblast cells during placentation. Tissue inhibitor of metalloproteinase-3 (TIMP-3), which is known as a potent inhibitor of MMP-9, also plays a role in regulating trophoblast invasion to the uterine endometrium. However little is known about the role of matrix metalloproteases and their inhibitors on regulation of maternal and fetal circulation. In the present study, we applied in vivo gene transfer system with hemagglutinating virus of Japan envelope (HVJ-E) vector into feto–maternal interface of pregnant mice on day 6.5 post coitus (pc), during the period of placental development. Overexpression of TIMP3 cDNA suppressedMMP-9 activity in the uterus. Maternal systolic blood pressure significantly rose from day 14.5 pc to parturition. The fetus and the placentae were significantly smaller, and serum soluble fms-like tyrosine kinase (Flt)-1 and soluble endoglin levelswere significantlyhigher after TIMP-3 treatment than in control mice. These data suggest that the balance between matrix metalloprotease and its inhibitor may play a pivotal role in determining feto/maternal circulation.

Secretion of cytokines by villous cytotrophoblast and extravillous trophoblast in the first trimester of human pregnancy

Cytokines are proposed to play roles in regulation of trophoblast invasion, spiral artery remodeling and immunoregulation during early pregnancy. Secretion of 12 cytokines (interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p70, IL-13, IFNγ, GM-CSF, MCP-1 and RANTES) by first trimester extravillous trophoblast and villous cytotrophoblast cells was examined using multiplex cytokine array technology. Seven (IL-1β, IL-8, IL-12p70, IL-13, GM-CSF, MCP-1 and RANTES) of the 12 cytokines examined were detectable in the samples studied (n=10 each group). Villous cytotrophoblast production of IL-1β and IL-8 increased with gestational age. Extravillous trophoblast production of IL-8, IL-13 and RANTES increased with gestational age. At 12-14 weeks gestation extravillous trophoblast cells secreted higher levels of IL-8, IL-13 and RANTES than villous cytotrophoblast cells.

The STOX1 genotype associated with pre-eclampsia leads to a reduction of trophoblast invasion by α-T-catenin upregulation

By using complementary in vitro and ex vivo approaches, we show that the risk allele (Y153H) of the pre-eclampsia susceptibility gene STOX1 negatively regulates trophoblast invasion by upregulation of the cell-cell adhesion protein alpha-T-catenin (CTNNA3). This is effectuated at the crucial epithelial-mesenchymal transition of proliferative into invasive extravillous trophoblast. This STOX1-CTNNA3 interaction is direct and includes Akt-mediated phosphorylated control of nucleo-cytoplasmic shuttling and ubiquitin-mediated degradation as shared with the FOX multigene family. This, to our knowledge, is the first time a genotype associated with pre-eclampsia has been shown to directly limit first trimester extravillous trophoblast invasion, the earliest hallmark of pre-eclampsia.