• AKT1
• in silico
• onset
• polymorphism
• preeclampsia
• severity

• Humans
• Proto-Oncogene Proteins c-akt/genetics
• Proto-Oncogene Proteins c-akt/metabolism
• Pre-Eclampsia/genetics
• Female
• Genetic Predisposition to Disease/genetics
• Polymorphism, Single Nucleotide/genetics
• Case-Control Studies
• Pregnancy
• Genotype
• Adult
• Alleles
• Risk Factors

• decidualization
• endometrial receptivity
• endometrium
• fibroids
• pregnancy

• Humans
• Female
• Leiomyoma
• Endometrium/pathology
• Pregnancy
• Uterine Neoplasms/pathology
• Embryo Implantation
• Signal Transduction
• Cytokines/metabolism

• endometrial receptivity
• glandular function
• glyphosate
• implantation failure
• proliferation

• PICT 2016 N° 0294 Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), Argentina
• PICT 2020 N°3506 Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), Argentina
• PICT 2020 N°3204 Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), Argentina
• PIP 437 2015 N° 11220150100338CO Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Argentina

• IVF
• PGT-A
• advanced maternal age
• blastocyst
• embryo quality
• implantation failure
• live birth
• miscarriage
• obesity
• trophectoderm biopsy

• Pregnancy
• Female
• Humans
• Abortion, Spontaneous
• Embryo Implantation
• Blastocyst
• Embryo Transfer/methods
• Genetic Testing/methods
• Retrospective Studies
• Aneuploidy
• Pregnancy Rate
• Preimplantation Diagnosis/methods

• Model organism
• Molecular genetics
• Phenotyping
• Pregnancy

• R21 HD088006 NICHD NIH HHS

• miRNA
• miscarriage
• pregnancy
• prenatal diagnosis
• preventive care

• Pregnancy
• Humans
• Female
• Abortion, Spontaneous/genetics
• MicroRNAs/genetics
• Pregnancy Complications
• Biomarkers

• genetics
• global genetic modification
• transgenic mice

• Female
• Male
• Mice
• Animals
• Recombinases
• Mice, Transgenic
• Receptors, Transforming Growth Factor beta
• Integrases/genetics
• Integrases/metabolism
• Protein Serine-Threonine Kinases

• ZIA ES103311 Intramural NIH HHS
• NIH

• cell growth
• decidualization
• endometrium
• phosphoinositide 3‐kinase signaling pathway
• platelet‐rich plasma

• Cre-LoxP
• HPG axis
• female infertility
• neuroendocrine-reproductive axis

Abnormal trophoblast behaviors during pregnancy contribute to the development of preeclampsia (PE). Syntaxin2 (STX2) has been shown to be a crucial epithelial mediator in numerous diseases. However, the functions of STX2 and the mechanisms underlying its role in PE remain largely unknown. The aim of this study was to explore the role of STX2 on trophoblast biology and unravel the molecular mechanisms that contribute to the development and progression of PE.

We first compared the expression of STX2 in placental tissues from women with PE and women with normal pregnancies. Then, we investigated the role of STX2 on trophoblast proliferation, migration and invasion in HTR-8/SVneo and primary human trophoblast cells by loss or gain of function experiments. In addition, co-immunoprecipitation, pulldown and immunofluorescence assays were performed to investigate the co-localization of STX2 with other proteins, and to help clarify the mechanisms underlying STX2-mediated functions on trophoblasts.

We demonstrated that STX2 expression was downregulated in placental tissues of women with PE compared with those from normal pregnancies. Loss and gain of function experiments further confirmed a role for STX2 in cell proliferation, migration and invasion in trophoblasts. By co-immunoprecipitation, pulldown and immunofluorescence co-localization assays, we revealed that STX2 selectively interacted with p85, a subunit of PI3K, and directly recruited p85 to the cytomembrane, thereby activating the AKT signaling pathway. We further demonstrated that the AKT activation was abolished by the use of a PI3K inhibitor (LY294002), which negatively affected STX2-mediated functions on trophoblasts.

All together, our findings point to a crucial role for STX2 in PE progression. Our new insights also suggest that STX2 may be a potential diagnostic tool and a novel therapeutic target for treating PE.

• AKT
• PI3K
• preeclampsia
• syntaxin2
• trophoblast

MicroRNAs (miRNAs) are widely expressed in different mammalian tissues and exert their biological effects through corresponding target genes. miRNA target genes can be rapidly and efficiently identified and screened by combining bioinformatics prediction and experimental validation. To investigate the possible molecular regulatory mechanisms involving miRNAs during uterine involution in postpartum ewes, we used Illumina HiSeq sequencing technology to screen for the number and characteristics of miRNAs in faster uterine involution and normal uterine involution group. A total of 118 differentially expressed miRNAs, including 33 known miRNAs and 85 new miRNAs, were identified in the hypothalamic library, whereas 54 miRNAs, including 5 known miRNAs and 49 new miRNAs, were identified in the uterine library. Screening with four types of gene prediction software revealed 73 target genes associated with uterine involution, and subsequently, GO annotation and KEGG pathway analysis were performed. The results showed that, in the hypothalamic–uterine axis, uterine involution in postpartum ewes might primarily involve two miRNA-target gene pairs, namely, miRNA-200a–PTEN and miRNA-133–FGFR1, which can participate in GnRH signal transduction in the upstream hypothalamus and in the remodeling process at the downstream uterus, through the PI3K–AKT signaling pathway to influence the recovery of the morphology and functions of the uterus during the postpartum period in sheep. Therefore, identification of differentially expressed miRNAs in this study fills a gap in the research related to miRNAs in uterine involution in postpartum ewes and provides an important reference point for a comprehensive understanding of the molecular mechanisms underlying the regulation of postpartum uterine involution in female livestock.

Abnormal birth weight is the one of the major causes of adulthood diseases such as obesity, metabolic syndrome, cardiovascular disease, type 2 diabetes, and hypertension. Accumulating evidence has suggested that the placental trophoblast is one of the most important reasons that influence birth weight. Our previous study showed that miR-519a are correlated with low fetal birth weight through regulating trophoblast proliferation. To further clarify the detailed mechanisms on how it is regulated, we screened the placental-specific circular RNAs (circRNAs) via microarray assay. The result identified that circ-SETD2 was highly expressed in the placenta of the patients with fetal macrosomia compared with healthy donors. Furthermore, bioinformatic analyses and the luciferase reporter assay revealed that miR-519a possessing the binding sites for both circ-SETD2 and phosphate and tensin homolog was deleted on chromosome 10 (PTEN). Interestingly, upregulation of circ-SETD2 enhanced the proliferation and invasion of the human trophoblast-like cell line HTR8/SVneo cell. A parallel study performed by Western blotting showed that overexpression of circ-SETD2 reduced miR-519a levels and increased PTEN levels in HTR8/SVneo cells. Importantly, the enhancement of HTR8/SVneo cell activity by circ-SETD2 overexpression was nullified when the cells were cotransfected by circ-SETD2 and miR-519a, suggesting the involvement of the circ-SETD2/miR-519a/PTEN axis in trophoblast activity. Taken together, we illustrate the role of circ-SETD2, as an upstream signaling of miR-519a/PTEN, in placenta development via regulating trophoblast proliferation and invasion. These findings improve our understanding of the mechanisms of progression of fetal macrosomia and will guide future development of therapeutic strategies against the disease by targeting the circ-SETD2/miR-519a/PTEN axis.

Kisspeptin-expressing neurons in the anteroventral periventricular nucleus (AVPV) and the arcuate nucleus (ARC) of the hypothalamus relay hormonal and metabolic information to gonadotropin-releasing hormone neurons, which in turn regulate pituitary and gonadal function. Phosphatase and tensin homolog (PTEN) blocks phosphatidylinositol 3-kinase (PI3K), a signaling pathway utilized by peripheral factors to transmit their signals. However, whether PTEN signaling in kisspeptin neurons helps to integrate peripheral hormonal cues to regulate gonadotropin release is unknown. To address this question, we generated mice with a kisspeptin cell-specific deletion of Pten (Kiss-PTEN KO), and first assessed kisspeptin protein expression and gonadotropin release in these animals. Kiss-PTEN KO mice displayed a profound sex and region-specific kisspeptin neuron hyperthrophy. We detected both kisspeptin neuron hyperthrophy as well as increased kisspeptin fiber densities in the AVPV and ARC of Kiss-PTEN KO females and in the ARC of Kiss-PTEN KO males. Moreover, Kiss-PTEN KO mice showed a reduced gonadotropin release in response to gonadectomy. We also found a hyperactivation of mTOR, a downstream PI3K target and central regulator of cell metabolism, in the AVPV and ARC of Kiss-PTEN KO females but not males. Fasting, known to inhibit hypothalamic kisspeptin expression and luteinizing hormone levels, failed to induce these changes in Kiss-PTEN KO females. We conclude that PTEN signaling regulates kisspeptin protein synthesis in both sexes and that its role as a metabolic signaling molecule in kisspeptin neurons is sex-specific.

• PTEN
• anteroventral periventricular nucleus
• arcuate nucleus
• hypothalamus
• kisspeptin
• luteinizing hormone
• mTOR

• Animals
• Arcuate Nucleus of Hypothalamus/metabolism
• Female
• Gene Targeting/methods
• Gonadotropin-Releasing Hormone/metabolism
• Gonadotropins/metabolism
• Humans
• Hypothalamus/metabolism
• Kisspeptins/genetics
• Kisspeptins/metabolism
• Luteinizing Hormone/metabolism
• Male
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Neurons/metabolism
• PTEN Phosphohydrolase/genetics
• PTEN Phosphohydrolase/metabolism
• Phosphatidylinositol 3-Kinases/metabolism
• Sex Factors
• Signal Transduction
• TOR Serine-Threonine Kinases/metabolism

• Animals
• Embryo Implantation/drug effects
• Female
• Genistein/adverse effects
• Mice
• Phytoestrogens/adverse effects
• Pregnancy
• Uterus/drug effects
• Uterus/growth & development
• Uterus/physiopathology

• ZIA ES102405 Intramural NIH HHS

• AKT
• ELA
• Invasion
• PI3K
• Preeclampsia

• AKT
• ROS
• Wnt
• cell cycle
• fetal membranes
• p38MAPK
• phosphorylation
• reproduction

• EPHB4
• Preeclampsia
• Trophoblast
• miR-454

• DMDD
• IUGR
• fetal growth restriction
• mouse models
• placenta
• trophoblast

• PI3K/PTEN/Akt signaling pathway
• endometrial receptivity
• immunohistochemistry
• intramural fibroids
• real-time PCR

• L-amino acid transporter-1 (lat1)
• Placenta
• mice

• AGC kinases
• AKT
• PDK1
• PI3K
• SGK1

Women with endometriosis can suffer from decreased fecundity or complete infertility via abnormal oocyte function or impaired placental-uterine interactions required for normal pregnancy establishment and maintenance. Although AT-rich interactive domain 1A (SWI-like) (ARID1A) is a putative tumor suppressor in human endometrial cancers and endometriosis-associated ovarian cancers, little is known about its role in normal uterine function. To study the potential function of ARID1A in the female reproductive tract, we generated mice with a conditional knockout of Arid1a using anti-Müllerian hormone receptor 2-Cre. Female Arid1a conditional knockout mice exhibited a progressive decrease in number of pups per litter, with a precipitous decline after the second litter. We observed no tumors in virgin mice, although one knockout mouse developed a uterine tumor after pregnancy. Unstimulated virgin female knockout mice showed normal oviductal, ovarian, and uterine histology. Uteri of Arid1a knockout mice showed a normal decidualization response and appropriate responses to estradiol and progesterone stimulation. In vitro studies using primary cultures of human endometrial stromal fibroblasts revealed that small interfering RNA knockdown of ARID1A did not affect decidualization in vitro. Timed pregnancy studies revealed the significant resorption of embryos at Embryonic Day 16.5 in knockout mice in the third pregnancy. In addition to evidence of implantation site hemorrhage, pregnant Arid1a knockout mice showed abnormal placental morphology. These results suggest that Arid1a supports successful pregnancy through its role in placental function.

• ARID1A
• female reproductive tract
• fertility
• genetically engineered mouse models
• placenta
• tumor suppressor
• uterus

Apoptosis is an important process in the reconstruction of endometrium within the menstrual cycle. The balance between cell proliferation and apoptosis regulates the periodic repair and shedding of endometrial cells and leads to the menstruation or prepare the mucosal layer of endometrium for the implantation of the embryo. Many factors with pro- and antiapoptotic action, such as B cell lymphoma/leukemia-2 and inhibitors apoptosis proteins families, caspases, tumor necrosis factor receptors, phosphatase and tensin homolog, proliferator-activated receptor gamma, microRNAs and others are differently expressed in the endometrial tissue at phases of menstrual cycle. Receptivity of the endometrium at the period of “window of implantation” is associated with the significant increase of apoptosis in endometrium to allow the embryo to be successfully implanted. The impairment of apoptosis regulation in the endometrium at this period often is observed in infertile women with endometriosis, tubal factor, polycystic ovary syndrome, etc.. In many cases the impairment of apoptosis regulation in the endometrium is the main cause of in vitro fertilization (IVF) treatment failure in these patients. As of today, the exact mechanisms and factors mediating the apoptotic process in normal endometrium and in infertile women are not fully understood. Herein, the literature data concerning the endometrial apoptosis regulation in general, and in light of the influence of apoptosis upon IVF treatment outcome are reviewed. The possibility to use some parameters of endometrial apoptosis for prediction of the successful pregnancy achievement in women participating in IVF protocols also is discussed.

• Apoptosis
• Endometrium
• Receptivity
• In vitro fertilization

• Apoptosis
• Invasion
• MicroRNA
• Proliferation
• Trophoblast
• miR-21

• Apoptosis
• Apoptosis Regulatory Proteins/genetics
• Apoptosis Regulatory Proteins/metabolism
• Cell Line, Transformed
• Cell Line, Tumor
• Cell Movement
• Cell Proliferation
• Female
• Gene Expression Regulation
• Humans
• MicroRNAs/antagonists & inhibitors
• MicroRNAs/genetics
• MicroRNAs/metabolism
• Oligonucleotides, Antisense/genetics
• Oligonucleotides, Antisense/metabolism
• Organ Specificity
• PTEN Phosphohydrolase/genetics
• PTEN Phosphohydrolase/metabolism
• Phosphorylation
• Pregnancy
• Primary Cell Culture
• Proto-Oncogene Proteins c-akt/genetics
• Proto-Oncogene Proteins c-akt/metabolism
• RNA-Binding Proteins/genetics
• RNA-Binding Proteins/metabolism
• Signal Transduction
• Transfection
• Trophoblasts/cytology
• Trophoblasts/metabolism

• Angiogenesis
• Decidualization
• Endometrium
• Gynecology
• Immunomodulation
• Implantation
• Pregnancy

Germline mutations of the Liver Kinase b1 (LKB1/STK11) tumor suppressor gene have been linked to Peutz-Jeghers Syndrome (PJS), an autosomal-dominant, cancer-prone disorder in which patients develop neoplasms in several organs, including the oviduct, ovary, and cervix. We have conditionally deleted Lkb1 in Müllerian duct mesenchyme-derived cells of the female reproductive tract and observed expansion of the stromal compartment and hyperplasia and/or neoplasia of adjacent epithelial cells throughout the reproductive tract with paratubal cysts and adenomyomas in oviducts and, eventually, endometrial cancer. Examination of the proliferation marker phospho-histone H3 and mammalian Target Of Rapamycin Complex 1 (mTORC1) pathway members revealed increased proliferation and mTORC1 activation in stromal cells of both the oviduct and uterus. Treatment with rapamycin, an inhibitor of mTORC1 activity, decreased tumor burden in adult Lkb1 mutant mice. Deletion of the genes for Tuberous Sclerosis 1 (Tsc1) or Tsc2, regulators of mTORC1 that are downstream of LKB1 signaling, in the oviductal and uterine stroma phenocopies some of the defects observed in Lkb1 mutant mice, confirming that dysregulated mTORC1 activation in the Lkb1-deleted stroma contributes to the phenotype. Loss of PTEN, an upstream regulator of mTORC1 signaling, along with Lkb1 deletion significantly increased tumor burden in uteri and induced tumorigenesis in the cervix and vagina. These studies show that LKB1/TSC1/TSC2/mTORC1 signaling in mesenchymal cells is important for the maintenance of epithelial integrity and suppression of carcinogenesis in adjacent epithelial cells. Because similar changes in the stromal population are also observed in human oviductal/ovarian adenoma and endometrial adenocarcinoma patients, we predict that dysregulated mTORC1 activity by upstream mechanisms similar to those described in these model systems contributes to the pathogenesis of these human diseases.

Peutz-Jeghers Syndrome patients have autosomal dominant mutations in the LKB1/STK11 gene and are prone to developing cancer, predominantly in the intestinal tract but also in other tissues, including the reproductive tracts and gonads. To elucidate the mechanisms disrupted by the loss of LKB1 in the reproductive tract, we have developed a mouse model with deletion of Lkb1 specifically in stromal cells of gynecologic tissues. These mice show stromal cell expansion and develop oviductal adenomas and endometrial cancer. Deletion of either Tsc1 or Tsc2 genes, which are mutated in patients with Tuberous Sclerosis Complex and whose protein products are indirect downstream targets of LKB1 signaling, resulted in some of the same defects observed in Lkb1 mutant mice. Activation of mammalian Target Of Rapamycin Complex 1 (mTORC1), a common effector of disrupted LKB1, TSC1, and TSC2 signaling, was observed in all mutant tissues examined, suggesting that uninhibited mTORC1 activity is necessary for the phenotypes. Suppression of mTORC1 signaling by rapamycin reduced tumor burden in Lkb1 mutant mice, confirming the link between dysregulation of mTORC1 to development of the Lkb1 mutant phenotype and suggesting that therapeutic targeting of LKB1/TSC1/TSC2/mTORC1 signaling would benefit human Peutz-Jeghers Syndrome and Tuberous Sclerosis patients with reproductive tract disease.

• AMP-Activated Protein Kinases
• Adenoma/drug therapy
• Adenoma/genetics
• Adenoma/pathology
• Animals
• Cervix Uteri/drug effects
• Cervix Uteri/metabolism
• Cervix Uteri/pathology
• Disease Models, Animal
• Endometrial Neoplasms/drug therapy
• Endometrial Neoplasms/genetics
• Endometrial Neoplasms/pathology
• Endometrium/drug effects
• Endometrium/metabolism
• Endometrium/pathology
• Female
• Gene Deletion
• Gene Expression Regulation, Neoplastic/drug effects
• Humans
• Mechanistic Target of Rapamycin Complex 1
• Mesenchymal Stem Cells/drug effects
• Mesenchymal Stem Cells/metabolism
• Mesenchymal Stem Cells/pathology
• Mice
• Multiprotein Complexes
• Ovary/drug effects
• Ovary/metabolism
• Ovary/pathology
• Oviducts/drug effects
• Oviducts/metabolism
• Oviducts/pathology
• PTEN Phosphohydrolase/deficiency
• PTEN Phosphohydrolase/genetics
• Protein Serine-Threonine Kinases/deficiency
• Protein Serine-Threonine Kinases/genetics
• Proteins/antagonists & inhibitors
• Proteins/genetics
• Signal Transduction/drug effects
• Sirolimus/administration & dosage
• TOR Serine-Threonine Kinases
• Tuberous Sclerosis Complex 1 Protein
• Tuberous Sclerosis Complex 2 Protein
• Tumor Microenvironment/drug effects
• Tumor Microenvironment/genetics
• Tumor Suppressor Proteins/deficiency
• Tumor Suppressor Proteins/genetics

• R01 HD052701 NICHD NIH HHS
• HD052701 NICHD NIH HHS