414. The contraceptive potential of a long-acting IL-11 inhibitor

2008 ◽  
Vol 20 (9) ◽  
pp. 94
Author(s):  
E. Menkhorst ◽  
L. Salamonsen ◽  
L. Robb ◽  
E. Dimitriadis
Keyword(s):  
Nature Medicine ◽  
Embryo Implantation ◽  
Hormonal Contraceptive ◽  
Cyclin D3 ◽  
Human Reproduction ◽  
Luminal Epithelium ◽  
Endometrial Stromal Cells ◽  
Decidual Cells ◽  
Implantation Sites ◽  
Long Acting

Interleukin 11 (IL-11) signalling is essential for the establishment of pregnancy in mice, through its action on the differentiation of uterine endometrial stromal cells (decidualisation), a critical process during embryo implantation. IL-11Rα deficient mice are infertile due to defective decidualisation1. IL-11 expression peaks between days (D) 4.5–9.5 of pregnancy (D0: day of plug) in mouse decidua. We examined the effect of administering (intraperitoneal [IP] injection or vaginal gel) a PEGylated IL-11 antagonist (PEGIL-11A) on decidualisation and pregnancy outcome in mice. The sera half-life of PEGIL-11A (IC50 2.8nM) following IP injection was 24h, compared with <1 h for the non-PEGylated antagonist (IC50 0.26nM). Following IP injection, PEGIL-11A localised to decidual cells and blocked the IL-11 decidual target protein, cyclin D3. IP injection of 600µg/application PEGIL-11A (or PEG control) at 1000 h and 1600 h on D3 and 1000 h on D4 (n = 4/group), resulted in smaller implantation sites than controls on D6 due to retarded mesometrial decidual formation. On D10, severe decidual destruction was visible: implantation sites contained regions of haemorrhage and the uterine luminal epithelium had reformed, suggesting a return to oestrous cycling. Following vaginal application in aqueous placebo gel, PEGIL-11A localised to decidual cells. Vaginal application of 200µg/application PEGIL-11A (or control) twice daily from D2 to D5 (n = 4/group), resulted in smaller implantation sites than controls on D6 due to partial inhibition of mesometrial decidual formation. This study demonstrates that PEGIL-11A blocked IL-11 action in the uterus, resulting in total pregnancy loss, equivalent to the IL-11Rα deficient mouse. In women, IL-11 and its receptor are produced by the uterine luminal and glandular epithelium during the period of uterine receptivity2, suggesting that IL-11 may act during initial blastocyst attachment to the luminal epithelium as well as stromal decidualisation. This study provides proof-of-principle for the development of a novel, non-hormonal contraceptive for women. (1) Robb L et al. Nature Medicine 1998; 4: 303–308. (2) Dimitriadis E et al. Molecular Human Reproduction 2000; 6: 907–914.

scholarly journals Possible roles of the cAMP-mediators EPAC and RAP1 in decidualization of rat uterus

Reproduction ◽  
2014 ◽  
Vol 147 (6) ◽  
pp. 897-906 ◽  
Author(s):  
Kazuya Kusama ◽  
Mikihiro Yoshie ◽  
Kazuhiro Tamura ◽  
Takiko Daikoku ◽  
Tsutomu Takarada ◽  
...  
Keyword(s):  
Embryo Implantation ◽  
Camp Signaling ◽  
Human Escs ◽  
Endometrial Stromal Cells ◽  
Camp Analog ◽  
Decidual Cells ◽  
Element Binding Protein ◽  
Implantation Sites ◽  
Responsive Element

The optimal decidualization of endometrial stromal cells (ESCs) following embryo implantation is one of the critical steps to establish pregnancy in rodents and humans. This step is intricately regulated by ovarian hormones. Using in vitro human ESCs model, we previously showed that activation of a cAMP mediator, exchange protein directly activated by cAMP (EPAC), promotes ovarian steroid- or cAMP analog-induced decidualization. However, expressions and functions of EPAC and RAP1 in the uterus during pregnancy have not yet been examined. In this study, we found that the expression of EPAC2 and RAP1 was markedly upregulated in the decidual cells at the implantation sites on days 7 and 9 of pregnancy in rats. Furthermore, both delayed-implantation and artificial decidualization models showed that EPAC2 and RAP1 expression was enhanced in decidual cells. Significant activation of cAMP-responsive element-binding protein (CREB), a central transcriptional factor of cAMP signaling, was observed in decidual cells. These spatiotemporal expressions of protein related EPAC pathway are overlapped by sites with activated cAMP signaling, indicating the association of EPAC signaling with decidualization. Strikingly, further studies in in vitro rat decidualization model showed that the cAMP analog and medroxyprogesterone stimulated the expression of decidual markers, while knockdown of EPAC1/2 and RAP1 attenuated the expressions of these markers. Together, these findings suggest that EPAC and RAP1 are the crucial factors for endometrial decidualization in rat pregnancy.

scholarly journals Cell–Cell Communication at the Embryo Implantation Site of Mouse Uterus Revealed by Single-Cell Analysis

2021 ◽  
Vol 22 (10) ◽  
pp. 5177
Author(s):  
Yi Yang ◽  
Jia-Peng He ◽  
Ji-Long Liu
Keyword(s):  
Single Cell ◽  
Molecular Mechanism ◽  
Single Cell Analysis ◽  
Embryo Implantation ◽  
Cell Communication ◽  
Human Reproduction ◽  
Mouse Uterus ◽  
Implantation Sites ◽  
Low Efficiency ◽  
Signaling Interactions

As a crucial step for human reproduction, embryo implantation is a low-efficiency process. Despite rapid advances in recent years, the molecular mechanism underlying embryo implantation remains poorly understood. Here, we used the mouse as an animal model and generated a single-cell transcriptomic atlas of embryo implantation sites. By analyzing inter-implantation sites of the uterus as control, we were able to identify global gene expression changes associated with embryo implantation in each cell type. Additionally, we predicted signaling interactions between uterine luminal epithelial cells and mural trophectoderm of blastocysts, which represent the key mechanism of embryo implantation. We also predicted signaling interactions between uterine epithelial-stromal crosstalk at implantation sites, which are crucial for post-implantation development. Our data provide a valuable resource for deciphering the molecular mechanism underlying embryo implantation.

scholarly journals The Role of Decidual Subpopulations in Implantation, Menstruation and Miscarriage

2021 ◽  
Vol 3 ◽  
Author(s):  
Joanne Muter ◽  
Chow-Seng Kong ◽  
Jan J. Brosens
Keyword(s):  
Stromal Cells ◽  
Acute Stress ◽  
Embryo Implantation ◽  
Recurrence Risk ◽  
Tissue Remodelling ◽  
Endometrial Stromal Cells ◽  
Decidual Cells ◽  
Unk Cells ◽  
Acute Stress Response

In each menstrual cycle, the endometrium becomes receptive to embryo implantation while preparing for tissue breakdown and repair. Both pregnancy and menstruation are dependent on spontaneous decidualization of endometrial stromal cells, a progesterone-dependent process that follows rapid, oestrogen-dependent proliferation. During the implantation window, stromal cells mount an acute stress response, which leads to the emergence of functionally distinct decidual subsets, reflecting the level of replication stress incurred during the preceding proliferative phase. Progesterone-dependent, anti-inflammatory decidual cells (DeC) form a robust matrix that accommodates the conceptus whereas pro-inflammatory, progesterone-resistant stressed and senescent decidual cells (senDeC) control tissue remodelling and breakdown. To execute these functions, each decidual subset engages innate immune cells: DeC partner with uterine natural killer (uNK) cells to eliminate senDeC, while senDeC co-opt neutrophils and macrophages to assist with tissue breakdown and repair. Thus, successful transformation of cycling endometrium into the decidua of pregnancy not only requires continuous progesterone signalling but dominance of DeC over senDeC, aided by recruitment and differentiation of circulating NK cells and bone marrow-derived decidual progenitors. We discuss how the frequency of cycles resulting in imbalanced decidual subpopulations may determine the recurrence risk of miscarriage and highlight emerging therapeutic strategies.

scholarly journals Exometabolomic analysis of decidualizing human endometrial stromal and perivascular cells

2020 ◽  
Author(s):  
Sarah Harden ◽  
Jieliang Zhou ◽  
Maria Diniz-da-Costa ◽  
Emma S. Lucas ◽  
Liang Cui ◽  
...  
Keyword(s):  
Time Course ◽  
Embryo Implantation ◽  
Cellular Reprogramming ◽  
Trophoblast Invasion ◽  
Perivascular Cells ◽  
Endometrial Stromal Cells ◽  
Signalling Molecules ◽  
Spatiotemporal Information ◽  
Decidual Cells ◽  
Acid Metabolites

ABSTRACTDifferentiation of endometrial fibroblasts into specialized decidual cells controls embryo implantation and transforms the cycling endometrium into a semi-permanent, immune-protective matrix that accommodates the placenta throughout pregnancy. This process starts during the midluteal phase of the menstrual cycle with decidual transformation of perivascular cells (PVC) surrounding the terminal spiral arterioles and endometrial stromal cells (EnSC) underlying the luminal epithelium. Decidualization involves extensive cellular reprogramming and acquisition of a secretory phenotype, essential for coordinated placental trophoblast invasion. Secreted metabolites are an emerging class of signalling molecules. Here, we used liquid chromatography-mass spectrometry to characterise the dynamic changes in metabolite secretion (exometabolome) of primary PVC and EnSC decidualized over 8 days. We identified 79 annotated metabolites differentially secreted upon decidualization, including prostaglandin, sphingolipid, and hyaluronic acid metabolites. Secreted metabolites encompassed 21 metabolic pathways, most prominently glycerolipid and pyrimidine metabolism. Although temporal exometabolome changes were comparable between decidualizing PVC and EnSC, 32 metabolites were differentially secreted across the decidualization time-course. Further, targeted metabolomics demonstrated a conspicuous difference in xanthine secretion between decidualized PVC and EnSC. Taken together, our findings indicate that the metabolic footprints generated by different decidual subpopulations encode spatiotemporal information that may be important for optimal embryo implantation.

172. DEVELOPMENT OF A NON-HORMONAL CONTRACEPTIVE: VAGINAL DELIVERY OF A LIF INHIBITOR: ITS TISSUE DISTRIBUTION AND EFFECT ON IMPLANTATION IN MICE

2009 ◽  
Vol 21 (9) ◽  
pp. 90
Author(s):  
E. M. Menkhorst ◽  
J. G. Zhang ◽  
P. O. Morgan ◽  
D. Metcalf ◽  
L. A. Salamonsen ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Tissue Distribution ◽  
Hormonal Contraceptive ◽  
Leukaemia Inhibitory Factor ◽  
Delivery Method ◽  
Time Points ◽  
Implantation Sites ◽  
Specific Tissue ◽  
Long Acting ◽  
Route Of Delivery

Uterine leukaemia inhibitory factor (LIF) is obligatory for fertility in mice and associated with infertility in women. Intraperitoneal injection (IPI) of a long-acting LIF inhibitor conjugated to polyethylene glycol (PEGLA) blocks endometrial LIF action preventing implantation1. Thus, PEGLA is a promising non-hormonal contraceptive. In women, vaginally applied (VA) compounds preferentially localize to the uterus suggesting a desirable delivery method for contraceptive purposes; however this has not been examined in mice. We aimed to compare the effects of VA and IPI PEGLA on tissue distribution and implantation in mice. Non-pregnant or mated female mice (Pregnant day [D] 2 [D0: day of plug], n=3/group) were given 125I-PEGLA by IPI (3x106cpm) or VA (7x105cpm). 125I-PEGLA was measured in blood and tissue at various time points. To block implantation, mated mice (n=4/group) were given PEGLA or control by IPI (500μg/injection) or VA (300μg/injection) at 1200h and 2200h on D2 and 1000h on D3 and the uterus examined for implantation sites on D6. 125 I-PEGLA accumulated in blood and uterus following IPI more rapidly (10min cf 30min), reached a higher concentration (10min, 6h and [blood] 24h; p<0.05) and remained longer (24h cf 6h) compared to VA. The percentage of protein-bound 125I in blood was higher following IPI (79.4+1.9%) than VA (47.5+6.7%) at 6h (p<0.05). Following IPI, 125I-PEGLA accumulated in the liver, gall bladder and stomach (2h) and spleen (24h) compared to control while no specific tissue accumulation was observed following VA. PEGLA prevented implantation following IPI (p<0.05) and reduced the size of implantation sites and decidualization in VA compared to controls. This study demonstrated differences in tissue distribution between VA and IPI PEGLA. It showed that VA PEGLA acted on the uterus in mice albeit to a lesser extent than IPI. It suggests that VA PEGLA is a potential route of delivery for contraceptive purposes.

Removal of Rev-erbα inhibition contributes to the prostaglandin G/H synthase 2 expression in rat endometrial stromal cells

2015 ◽  
Vol 308 (8) ◽  
pp. E650-E661 ◽  
Author(s):  
Keishiro Isayama ◽  
Lijia Zhao ◽  
Huatao Chen ◽  
Nobuhiko Yamauchi ◽  
Yasufumi Shigeyoshi ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Chromatin Immunoprecipitation ◽  
Clock Genes ◽  
Pcr Analysis ◽  
Core Component ◽  
Endometrial Stromal Cells ◽  
Transcript Levels ◽  
Rhythmic Expression ◽  
Decidual Cells ◽  
Implantation Sites

The rhythmic expression of clock genes in the uterus is attenuated during decidualization. This study focused on Ptgs2, which is essential for decidualization, as a putative clock-controlled gene, and aimed to reveal the functions of clock genes in relation to Ptgs2 during decidualization. We compared the transcript levels of clock genes in the rat uterus on days 4.5 (D4.5) and 6.5 of pregnancy. The transcript levels of clock genes ( Per2, Bmal1, Rorα, and Rev-erbα) had decreased at implantation sites on day 6.5 (D6.5e) compared with those on D4.5, whereas Ptgs2 transcripts had increased on D6.5e. Similar observations of Rev-erbα and Ptgs2 were also obtained in the endometrium on D6.5e by immunohistochemistry. In the decidual cells induced by medroxyprogesterone and 2- O-dibutyryl-cAMP, the rhythmic expression levels of clock genes were attenuated, whereas Ptgs2 transcription was induced. These results indicate that decidualization causes the attenuation of clock genes and the induction of Ptgs2. Furthermore, in the experiment of Bmal1 siRNA, the rhythmic expression of clock genes and Ptgs2 was attenuated by the siRNA. Transcript levels of Ptgs2 and prostaglandin (PG)E2 production were increased by treatment with the Rev-erbα antagonist, suggesting the contribution of the nuclear receptor Rev-erbα to Ptgs2 expression. Moreover, Rev-erbα knockdown enhanced the induction of Ptgs2 transcription and PGE2 production by forskolin. Chromatin immunoprecipitation-PCR analysis revealed that Rev-erbα could directly bind to a proximal RORE site of Ptgs2. Collectively, this study demonstrates that the attenuation of the circadian clock, especially its core component Rev-erbα, contributes to the induction of Ptgs2 during decidualization.

scholarly journals 227.An inhibitor of leukemia inhibitory factor signalling blocks embryo implantation in the mouse

2004 ◽  
Vol 16 (9) ◽  
pp. 227
Author(s):  
E. Dimitriadis ◽  
C. Stoikos ◽  
M. Baca ◽  
W. Fairlie ◽  
A. D. Uboldi ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Leukemia Inhibitory Factor ◽  
Embryo Implantation ◽  
Uterine Horn ◽  
Inhibitory Factor ◽  
Luminal Epithelium ◽  
Pregnant Uterus ◽  
Implantation Sites ◽  
Post Implantation

Embryo implantation is a critical step in the establishment of pregnancy. Endometrial leukemia inhibitory factor (LIF) is essential for embryo implantation in the mouse (1). Uterine LIF is expressed in the luminal epithelium on Day 3 of pregnancy (D3) (D0�=�day of plug detection) and signals via activation of signal transducer and activator of transcription (Stat) 3 (2). We examined the effect of a novel LIF signalling inhibitor on the phosphorylation (p) of Stat3 during early pregnancy and on embryo implantation in the mouse. We injected LIF inhibitor into one uterine horn and PBS into the other uterine horn of the mouse at D3 and examined the effect on pStat3 immunostaining in the luminal epithelium between 30 and 360�min later. We found no immunoreactive pStat3 in luminal epithelium following treatment with LIF inhibitor at 60 and 90�min but variable staining at other time points. The PBS-treated uterine horn showed intense immunostaining at all times. LIF inhibitor (1mg/kg body weight per day) or PBS was administered to mice (a) subcutaneously, (b) intraperitoneally, at 8-hourly intervals for 3�days from D2, or (c) continuously into the peritoneal cavity via Alzet pumps from D2. No effect was seen on implantation at D6. When LIF antagonist (3.5mg/kg/day) or PBS were administered by Alzet pumps from D2 together with ip injections, 4-hourly from D3 for 36�h, there were no implantation sites in the uteri of treated mice (n�=�5) while the control mice (n�=�4) had 3.6���0.5�sites (P�<�0.001). Histologically, the uteri of the treated mice resembled non-pregnant uterus, while the control uterus resembled post-implantation uterus. The results demonstrate that treatment of mice during early pregnancy with a novel LIF inhibitor blocks LIF action in vivo and embryo implantation. This knowledge is important for development of novel contraceptives. (1) Stewart, C. L., Kaspar, P., Brunet, L. J., Bhatt, H., Gadi, I., Kontgen, F., Abbondanzo, S. J. (1992) Nature 359, 76–79. (2) Cheng, J. G., Chen, J. R., Hernandez, L., Alvord, W. G., Stewart, C. L. (2001) Proc. Natl Acad. Sci. USA 98, 8680–8685.

scholarly journals The Androgen and Progesterone Receptors Regulate Distinct Gene Networks and Cellular Functions in Decidualizing Endometrium

Endocrinology ◽  
2008 ◽  
Vol 149 (9) ◽  
pp. 4462-4474 ◽  
Author(s):  
Brianna Cloke ◽  
Kaisa Huhtinen ◽  
Luca Fusi ◽  
Takeshi Kajihara ◽  
Maria Yliheikkilä ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Gene Networks ◽  
Cell Cycle Regulation ◽  
Embryo Implantation ◽  
Fiber Formation ◽  
Cellular Functions ◽  
Endometrial Stromal Cells ◽  
Cytoskeletal Organization ◽  
Decidual Cells ◽  
Cycle Regulation

Progesterone is indispensable for differentiation of human endometrial stromal cells (HESCs) into decidual cells, a process that critically controls embryo implantation. We now show an important role for androgen receptor (AR) signaling in this differentiation process. Decreased posttranslational modification of the AR by small ubiquitin-like modifier (SUMO)-1 in decidualizing cells accounted for increased responsiveness to androgen. By combining small interfering RNA technology with genome-wide expression profiling, we found that AR and progesterone receptor (PR) regulate the expression of distinct decidual gene networks. Ingenuity pathway analysis implicated a preponderance of AR-induced genes in cytoskeletal organization and cell motility, whereas analysis of AR-repressed genes suggested involvement in cell cycle regulation. Functionally, AR depletion prevented differentiation-dependent stress fiber formation and promoted motility and proliferation of decidualizing cells. In comparison, PR depletion perturbed the expression of many more genes, underscoring the importance of this nuclear receptor in diverse cellular functions. However, several PR-dependent genes encode for signaling intermediates, and knockdown of PR, but not AR, compromised activation of WNT/β-catenin, TGFβ/SMAD, and signal transducer and activator of transcription (STAT) pathways in decidualizing cells. Thus, the nonredundant function of the AR in decidualizing HESCs, centered on cytoskeletal organization and cell cycle regulation, implies an important role for androgens in modulating fetal-maternal interactions. Moreover, we show that PR regulates HESC differentiation, at least in part, by reprogramming growth factor and cytokine signal transduction.

scholarly journals Hoxa-10 and Cyclin D3 Overexpression in the Decidual Reaction in a Superovulation Protocol in Young Adult C57BL/6J Mice

2016 ◽  
Vol 54 (2) ◽  
pp. 328-335 ◽  
Author(s):  
T. Garcia Robles ◽  
R. A. García Fernández ◽  
P. García-Palencia ◽  
M. D. Arrabal ◽  
B. Sánchez Maldonado ◽  
...  
Keyword(s):  
Growth Factor ◽  
Stromal Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Cyclin D3 ◽  
Proliferation Index ◽  
Control Group ◽  
Ki 67 ◽  
Endometrial Stromal Cells ◽  
Decidual Cells ◽  
Decidual Reaction

Following the performance of a superovulation protocol, multiple nodules were observed bilaterally in the uterine horns of 31 of 276 (11.2%) C57BL/6 J female mice aged 8.5 ± 0.6 (mean and standard error of mean) weeks. These lesions prevented embryo collection, and the uterine decidual reaction was suspected. Samples of pathological uteri (n = 20) and the normal genital tracts of donors treated with a similar superovulation protocol (control group, n = 10) were collected. Immunohistochemistry was performed to evaluate pancytokeratin, desmin, vimentin, progesterone receptor (PR), estrogen receptor α (ERα), Ki-67, cyclin D3 and c-Myc expression, as well as quantitative polymerase chain reaction to assess cyclin D3, Hoxa-10 and heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA expression. The uterine decidual reaction presented a high degree of structural organization and specifically affected the antimesometrial region of the endometrium. The abnormal decidual cells were large polygonal cells that were frequently polyploid or binucleated and strongly positive for desmin. Immunohistochemistry showed higher Ki-67 proliferation index and higher expression of PR and cyclin D3 in decidual cells in the antimesometrial aspect of the endometrium, compared to nondecidualized endometrial stromal cells in the mesometrial aspect of affected uteri, and compared to endometrial stromal cells in healthy uteri. High expression of cyclin D3 and Hoxa-10 mRNA was also observed in uteri affected by the decidual reaction. These results suggest that PR overexpression in endometrial stromal cells, likely due to high progesterone levels, triggers cyclin D3 and Hoxa-10 overexpression, which may be involved in the pathological mechanisms of the mouse uterine decidual reaction.

scholarly journals Identification of Intercellular Crosstalk between Decidual Cells and Niche Cells in Mice

2021 ◽  
Vol 22 (14) ◽  
pp. 7696
Author(s):  
Jia-Peng He ◽  
Qing Tian ◽  
Qiu-Yang Zhu ◽  
Ji-Long Liu
Keyword(s):  
Molecular Mechanism ◽  
Immune Cells ◽  
Embryo Implantation ◽  
Global Gene Expression ◽  
Human Reproduction ◽  
Valuable Resource ◽  
Cell Type ◽  
Mouse Uterus ◽  
Crucial Step ◽  
Decidual Cells

Decidualization is a crucial step for human reproduction, which is a prerequisite for embryo implantation, placentation and pregnancy maintenance. Despite rapid advances over recent years, the molecular mechanism underlying decidualization remains poorly understood. Here, we used the mouse as an animal model and generated a single-cell transcriptomic atlas of a mouse uterus during decidualization. By analyzing the undecidualized inter-implantation site of the uterus as a control, we were able to identify global gene expression changes associated with decidualization in each cell type. Additionally, we identified intercellular crosstalk between decidual cells and niche cells, including immune cells, endothelial cells and trophoblast cells. Our data provide a valuable resource for deciphering the molecular mechanism underlying decidualization.

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