scholarly journals Loss of imprinting control of the lncRNA H19-fetal mitogen IGF2 gene cluster in the decidual microenvironment of patients with idiopathic spontaneous miscarriages

2021 ◽  
Author(s):  
Xue Wen ◽  
Qi Zhang ◽  
Lei Zhou ◽  
Zhaozhi Li ◽  
Xue Wei ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Noncoding Rna ◽  
Recurrent Miscarriage ◽  
Critical Role ◽  
Ctcf Binding ◽  
Ctcf Binding Site ◽  
Monoallelic Expression ◽  
Endometrial Stromal Cells ◽  
Control Center ◽  
Loss Of Imprinting

Abstract Miscarriage, the spontaneous loss of a pregnancy before the fetus achieves viability, is a common complication of pregnancy. Decidualization plays a critical role in the implantation of the embryo. To search for molecular factors underlying miscarriage, we explored the role of long noncoding RNAs (lncRNAs) in the decidual microenvironment, where the molecular crosstalk at the feto–maternal interface occurs. By integrating RNA-seq data from recurrent miscarriage patients and decidualized endometrial stromal cells, we identified H19 , a noncoding RNA that exhibits paternally imprinted monoallelic expression in normal tissues, as the most upregulated lncRNA associated with miscarriage. Aberrant upregulation of H19 lncRNA was observed in decidual tissues derived from patients with spontaneous miscarriage as well as decidualized endometrial stromal cells. The maternally imprinted fetal mitogen Igf2, which is usually reciprocally co-regulated with H19 in the same imprinting cluster, was also upregulated. Notably, both genes underwent loss of imprinting, as H19 and IGF2 were actively transcribed from both parental alleles in decidual tissues. Mechanistically, this loss of imprinting in decidual tissues was associated with the loss of the H3K27m3 suppression marker in the IGF2 promoter, CpG hypomethylation at the central CTCF binding site in the imprinting control center (ICR) that is located between IGF2 and H19 , and the loss of CTCF-mediated intrachromosomal looping. These data provide the first evidence that aberrant control of the ICR epigenotype-intrachromosomal looping- H19/IGF2 imprinting pathway may be a critical epigenetic risk factor in the abnormal decidualization related to miscarriage.

scholarly journals Downregulation of decidual SKP2 is associated with human recurrent miscarriage

2021 ◽  
Author(s):  
Shijian Lv ◽  
Mei Liu ◽  
Lizhen Xu ◽  
Cong Zhang
Keyword(s):  
Stromal Cells ◽  
Recurrent Miscarriage ◽  
Aerobic Glycolysis ◽  
Critical Role ◽  
Protein S ◽  
Pcr Analysis ◽  
Endometrial Stromal Cells ◽  
Downstream Target ◽  
F Box Protein

Abstract Background: Recurrent miscarriage (RM) is a very frustrating problem for both couples and clinicians. To date, the etiology of RM remains poorly understood. Decidualization plays a critical role in implantation and the maintenance of pregnancy, and its deficiency is closely correlated with RM. The F-box protein S-phase kinase associated protein 2 (SKP2) is a key component of the SCF-type E3 ubiquitin ligase complex, which is critically involved in ErbB family-induced Akt ubiquitination, aerobic glycolysis and tumorigenesis. SKP2 is pivotal for reproduction, and SKP2-deficient mice show impaired ovarian development and reduced fertility.Methods: Here, we investigated the expression and function of SKP2 in human decidualization and its relation with RM. A total of 40 decidual samples were collected. Quantitative PCR analysis, western blot analysis and immunohistochemistry analysis were performed to analyze the differential expression of SKP2 between RM and control cells. For in vitro induction of decidualization, both HESCs (human endometrial stromal cells) cell line and primary ESCs (endometrial stromal cells) were used to analyze the effects of SKP2 on decidualization via siRNA transfection.Results: Compared to normal pregnant women, the expression of SKP2 was reduced in the decidual tissues from individuals with RM. After in vitro induction of decidualization, knockdown of SKP2 apparently attenuated the decidualization of HESCs and resulted in the downregulation of HOXA10 and FOXM1, which are essential for normal human decidualization. Moreover, our experiments demonstrated that SKP2 silencing reduced the expression of its downstream target GLUT1.Conclusions: Our study indicates a functional role of SKP2 in RM: downregulation of SKP2 in RM leads to impaired decidualization and downregulation of GLUT1 and consequently predisposes individuals to RM.

scholarly journals Downregulation of decidual SKP2 is associated with human recurrent miscarriage

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Shijian Lv ◽  
Mei Liu ◽  
Lizhen Xu ◽  
Cong Zhang
Keyword(s):  
Stromal Cells ◽  
Recurrent Miscarriage ◽  
Aerobic Glycolysis ◽  
Critical Role ◽  
Protein S ◽  
Pcr Analysis ◽  
Endometrial Stromal Cells ◽  
Downstream Target ◽  
In Vitro Induction

Abstract Background Recurrent miscarriage (RM) is a very frustrating problem for both couples and clinicians. To date, the etiology of RM remains poorly understood. Decidualization plays a critical role in implantation and the maintenance of pregnancy, and its deficiency is closely correlated with RM. The F-box protein S-phase kinase associated protein 2 (SKP2) is a key component of the SCF-type E3 ubiquitin ligase complex, which is critically involved in ErbB family-induced Akt ubiquitination, aerobic glycolysis and tumorigenesis. SKP2 is pivotal for reproduction, and SKP2-deficient mice show impaired ovarian development and reduced fertility. Methods Here, we investigated the expression and function of SKP2 in human decidualization and its relation with RM. A total of 40 decidual samples were collected. Quantitative PCR analysis, western blot analysis and immunohistochemistry analysis were performed to analyze the differential expression of SKP2 between RM and control cells. For in vitro induction of decidualization, both HESCs (human endometrial stromal cells) cell line and primary ESCs (endometrial stromal cells) were used to analyze the effects of SKP2 on decidualization via siRNA transfection. Results Compared to normal pregnant women, the expression of SKP2 was reduced in the decidual tissues from individuals with RM. After in vitro induction of decidualization, knockdown of SKP2 apparently attenuated the decidualization of HESCs and resulted in the downregulation of HOXA10 and FOXM1, which are essential for normal human decidualization. Moreover, our experiments demonstrated that SKP2 silencing reduced the expression of its downstream target GLUT1. Conclusions Our study indicates a functional role of SKP2 in RM: downregulation of SKP2 in RM leads to impaired decidualization and downregulation of GLUT1 and consequently predisposes individuals to RM.

301. LEUKEMIA INHIBITORY FACTOR IS A CRITICAL REGULATOR OF DECIDUALIZATION OF ENDOMETRIAL STROMAL CELLS IN HUMANS AND MICE

2010 ◽  
Vol 22 (9) ◽  
pp. 101
Author(s):  
L. Lin ◽  
E. M. Menkhorst ◽  
E. Dimitriadis
Keyword(s):  
Stromal Cells ◽  
Leukemia Inhibitory Factor ◽  
Recurrent Miscarriage ◽  
Embryo Implantation ◽  
Critical Role ◽  
Prolactin Secretion ◽  
Inhibitory Factor ◽  
Endometrial Stromal Cells ◽  
Decidual Cells

Decidualization is the differentiation of endometrial stromal cells into decidual cells. It is a critical process in embryo implantation, placentation and the establishment of pregnancy. Inadequate decidualization can lead to infertility, abnormal placentation and recurrent miscarriage. Endometrial leukemia inhibitory factor (LIF) is indispensible in blastocyst implantation in mice and dysregulated in infertile women. LIF is produced by 1st trimester decidual cells but its role in decidualization is not known. This study aimed to examine the role of LIF in human and mouse decidualization. Primary human endometrial stomal cells (HESC) were isolated and decidualized (D) by treatment with estradiol (E) +medroxyprogesterone acetate (MPA) for 14 days. HESC were also treated with E+MPA+/–LIF (0.5, 5, 50, 100 and 200 ng/mL) for 14 days. Prolactin secretion was used to assess the extent of decidualization (n = 6). D and non-D HESC were also treated with LIF (0.5, 5, 50, 100 and 200 ng/mL +/– LIF inhibitor) for 15min and the phosphorylation (p) of signal transducer and activator of transcription (STAT)3/STAT3 abundance was detected by Western blot (n = 4). RNA was isolated for analysis of LIF and LIF receptor (R) mRNA expression during decidualization (n = 4). HESC treated with E+MPA+LIF (50, 100 and 200 ng/mL) secreted more prolactin compared to cells treated with E+MPA alone (P < 0.05). LIF increased pSTAT3/STAT3 abundance in D and non-D cells while LIF+LIF inhibitor abolished pSTAT3/STAT3. LIF mRNA was downregulated while LIF-R mRNA increased during decidualization. In vivo, mated mice (n = 5) were injected intraperitoneally with a unique long acting LIF inhibitor post-implantation at day 4.5 of pregnancy and resulted in reduced decidualization compared to control. This is the first study to demonstrate that LIF promoted decidualization of HESC possibly via pSTAT3. It further suggested that LIF regulated decidualization in mice demonstrating a newly identified critical role for LIF in the establishment of pregnancy.

scholarly journals Critical Role of TRPC1-Mediated Ca2+Entry in Decidualization of Human Endometrial Stromal Cells

2012 ◽  
Vol 26 (5) ◽  
pp. 846-858 ◽  
Author(s):  
Yasuhiro Kawarabayashi ◽  
Lin Hai ◽  
Akira Honda ◽  
Shinji Horiuchi ◽  
Hiroshi Tsujioka ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Critical Role ◽  
Endometrial Stromal Cells

scholarly journals Activated Hippo/Yes-Associated Protein Pathway Promotes Cell Proliferation and Anti-apoptosis in Endometrial Stromal Cells of Endometriosis

2016 ◽  
Vol 101 (4) ◽  
pp. 1552-1561 ◽  
Author(s):  
Yong Song ◽  
Jing Fu ◽  
Min Zhou ◽  
Li Xiao ◽  
Xue Feng ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Signaling Pathway ◽  
Nude Mice ◽  
Stromal Cells ◽  
Target Genes ◽  
Critical Role ◽  
B Cell Lymphoma ◽  
Endometrial Stromal Cells ◽  
Proliferation And Apoptosis

Abstract Context: The imbalance in cell proliferation and apoptosis is considered an important role in the pathogenesis of endometriosis, but the exact mechanisms remains unclear. A newly established signaling pathway–Hippo/Yes-associated protein (YAP) pathway plays a critical role in the proliferation and apoptosis processes. However, studies focusing on Hippo/YAP pathway and endometriosis are lacking. Objective: The objective was to explore the function of the Hippo/YAP pathway in endometriosis. Setting and Design: The expression of YAP was first investigated in endometrium of women with or without endometriosis. The role of YAP in cell proliferation and apoptosis is identified by transfection of endometrial stromal cells (ESCs) in vitro, subsequent Verteporfin treatments in eutopic ESCs in vitro, and endometriosis animal model of nude mice in vivo. Results: Our results revealed that increased expression of YAP and decreased expression of p-YAP in ectopic and eutopic endometrium compared with normal endometrium. YAP knockdown in eutopic ESCs decreased cell proliferation and enhanced cell apoptosis companied with decreased expression of TEAD1, CTGF, and B-cell lymphoma/leukemia (BCL)-2; whereas overexpression of YAP resulted in increased proliferation and decreased apoptosis of normal ESCs with increased expression of TEAD1, CTGF, and BCL-2. By chromatin immunoprecipitation qPCR CTGF and BCL-2 were identified as directly downstream target genes of YAP-TEAD1 active complex. Eutopic ESCs treated with Verteporfin revealed decreased proliferation and enhanced apoptosis whereas in endometriosis animal models of nude mice treated with Verteporfin, the size of endometriotic lesions was significantly reduced. Conclusions: Our study suggests that the Hippo/YAP-signaling pathway plays a critical role in the pathogenesis of endometriosis and should present a novel therapeutic method against endometriosis.

scholarly journals Hypoxia-Induced MicroRNA-20a Expression Increases ERK Phosphorylation and Angiogenic Gene Expression in Endometriotic Stromal Cells

2012 ◽  
Vol 97 (8) ◽  
pp. E1515-E1523 ◽  
Author(s):  
Shih-Chieh Lin ◽  
Chih-Chuan Wang ◽  
Meng-Hsing Wu ◽  
Shang-Hsun Yang ◽  
Yo-Hua Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stromal Cells ◽  
Noncoding Rna ◽  
Hypoxia Inducible Factor ◽  
Transcriptional Response ◽  
Fine Tuning ◽  
Translational Efficiency ◽  
Endometrial Stromal Cells ◽  
Small Noncoding Rna ◽  
Erk Phosphorylation

Abstract Context: Aberrant activation of MAPK has been implicated to play important roles in pathological processes of endometriosis. However, how MAPK are constitutively activated in endometriotic tissues remains largely unknown. microRNA are small noncoding RNA that regulate the stability or translational efficiency of target mRNA by interacting with the 3′ untranslated region. Thus, miRNA are thought to be modulators of the transcriptional response, fine-tuning gene expression. Objective: The aim of this study was to evaluate the functional roles of microRNA-20a (miR20a) in MAPK activation and the pathogenesis of endometriosis. Design: miR20a expression was analyzed in nonpaired (endometrium = 17; endometriosis = 37) and paired (n = 12) endometriotic tissues by quantitative RT-PCR. Overexpression of miR20a in eutopic endometrial stromal cells or inhibition of miR20a in ectopic endometriotic stromal cells was used to evaluate its impact on ERK phosphorylation and subsequently angiogenesis- and proliferation-related gene expression. Results: Levels of miR20a were up-regulated in endometriotic stromal cells. Elevation of miR20a was up-regulated by hypoxia inducible factor-1α. The up-regulation of miR20a causes the down-regulation of dual-specificity phosphatase-2, which leads to prolonged ERK phosphorylation and an increase in the expression of several angiogenic genes. Furthermore, the up-regulation of miR20a enhances the prostaglandin E2-induced expression of fibroblast growth factor-9, a potent mitogen that stimulates both endothelial and endometrial cell proliferation. Conclusion: Our findings provide the novel mechanism that not only functionally links together hypoxic stress, miR20a expression, aberrant ERK phosphorylation, and angiogenesis but also demonstrates that miR20a is an important modulator in the development of endometriosis.

scholarly journals Endometrial Stromal Cells of Women with Recurrent Miscarriage Fail to Discriminate between High- and Low-Quality Human Embryos

PLoS ONE ◽  
2012 ◽  
Vol 7 (7) ◽  
pp. e41424 ◽  
Author(s):  
Charlotte H. E. Weimar ◽  
Annemieke Kavelaars ◽  
Jan J. Brosens ◽  
Birgit Gellersen ◽  
Johanna M. T. de Vreeden-Elbertse ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Recurrent Miscarriage ◽  
Human Embryos ◽  
Endometrial Stromal Cells

scholarly journals Loop competition and extrusion model predicts CTCF interaction specificity

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Wang Xi ◽  
Michael A. Beer
Keyword(s):  
Binding Site ◽  
Critical Role ◽  
Three Dimensional ◽  
Ctcf Binding ◽  
Chromatin Interaction ◽  
Ctcf Binding Site ◽  
Loop Formation ◽  
Critical Determinant ◽  
Frequency Changes ◽  
Transcriptional Gene Regulation

AbstractThree-dimensional chromatin looping interactions play an important role in constraining enhancer–promoter interactions and mediating transcriptional gene regulation. CTCF is thought to play a critical role in the formation of these loops, but the specificity of which CTCF binding events form loops and which do not is difficult to predict. Loops often have convergent CTCF binding site motif orientation, but this constraint alone is only weakly predictive of genome-wide interaction data. Here we present an easily interpretable and simple mathematical model of CTCF mediated loop formation which is consistent with Cohesin extrusion and can predict ChIA-PET CTCF looping interaction measurements with high accuracy. Competition between overlapping loops is a critical determinant of loop specificity. We show that this model is consistent with observed chromatin interaction frequency changes induced by CTCF binding site deletion, inversion, and mutation, and is also consistent with observed constraints on validated enhancer–promoter interactions.

scholarly journals Integrated analysis of long non-coding RNA and mRNA expression in endometrial stromal cells induced by poly (I:C) identifies immune response genes

2020 ◽  
Author(s):  
Ming Zhang ◽  
Yan Zhang ◽  
Qiuying Wu ◽  
Ling Xu ◽  
Yutian Zeng ◽  
...  
Keyword(s):  
Immune Response ◽  
Virus Infection ◽  
Signaling Pathways ◽  
Expression Analysis ◽  
Stromal Cells ◽  
Noncoding Rna ◽  
Expression Profiles ◽  
Poly I:C ◽  
Integrated Analysis ◽  
Endometrial Stromal Cells

Abstract Background The uterus of an animal is relatively easily infected by pathogenic microorganisms, which can cause serious reproductive disorders and economic loss to animal husbandry. The presence of long noncoding RNA (lncRNA) is closely related to many diseases. Poly(I:C) is a synthetic double-stranded RNA that is often used as a substitute for dsRNA viral infection. In this study, we analyzed the mRNA and lncRNA expression profiles of in vitro cultured rabbit endometrial stromal cells (ESCs) after poly(I:C)-induction, to explore the role of these RNAs in the immune response. Results We identified 10,927 lncRNAs and 20,494 mRNAs, of which 291 lncRNAs and 1311 mRNAs were significantly differentially expressed (DE) between the control and poly(I:C) groups ( p <0.05). GO and KEGG analysis showed that DE genes and target genes of DE lncRNAs were enriched in relation to the occurrence of various diseases, development of tissues and organs, metabolic processes, and the immune response. Moreover, these genes were also enriched in many pathways related to immune and inflammatory responses, such as the toll-like receptor and the NF-κB and Jak-STAT signaling pathways. Co-expression analysis of lncRNA and mRNA revealed that there were significant relationships between a number of lncRNAs including MSTRG.153189.1, MSTRG.102664.8, MSTRG.39626.1, MSTRG.68469.1, MSTRG.137189.4, MSTRG.32118.5 and MSTRG.76080.1, and the immune system genes, CCL2, CCL5, IL-1, IL-6, IFN and ISG15, which suggested that lncRNAs in ESCs might be involved in regulation of the immune response to poly(I:C) through genes related to immune signaling pathways. Conclusions Our results provide both transcriptomic and epigenetic insights into the immune response of uterine cells to dsRNA virus infection. Comprehensive lncRNA and mRNA transcriptomes in rabbit ESCs exposed to poly(I:C) were profiled. Co-expression analysis identified an integrated lncRNA-mRNA interaction network, implying that key genes or lncRNAs exerted critical influences on the immune response to virus infection.

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