scholarly journals Identifying stably expressed housekeeping genes in the endometrium of fertile women, women with recurrent implantation failure and recurrent miscarriages

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Linden Stocker ◽  
Felino Cagampang ◽  
Ying Cheong
Keyword(s):  
Housekeeping Genes ◽  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
Recurrent Miscarriages ◽  
Fertile Women

P–388 Endometrial extracellular vesicles from recurrent implantation failure patients inhibited embryonic growth and implantation via miR–6131/PAK2 pathway

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Liu
Keyword(s):  
Extracellular Vesicles ◽  
Migration And Invasion ◽  
Hatching Rate ◽  
Implantation Failure ◽  
Trophoblast Cells ◽  
Recurrent Implantation Failure ◽  
Endometrial Cells ◽  
Blastocyst Rate ◽  
Fertile Women

Abstract Study question Could endometrial extracellular vesicles from recurrent implantation failure patients (RIF-EVs) attenuate the growth and implantation potentials of embryos and what are the mechanisms? Summary answer: RIF-EVs inhibited embryonic growth and decreased the trophoblast functions via miR–6131/PAK2 pathway. What is known already Recurrent implantation failure (RIF) is characterized by repeated embryo transfers without pregnancy. To date, the etiology of RIF remains poorly understood. Recent evidence indicated that extracellular vesicles (EVs) secreted by endometrial cells, played a crucial role in the implantation by regulating the development and implantation of embryos. Study design, size, duration Endometrial cells isolated from endometrial tissues of RIF patients (n = 25) and fertile women (n = 16) were cultured and modulated via hormones. Endometrial EVs from RIF patients (RIF-EVs) or fertile women (FER-EVs) were isolated from the conditioned medium. The influence of EVs on embryonic development and implantation was investigated by co-culture models of EVs and 2-cell murine embryos or HTR8/SVneo cells, respectively. High-through put sequencing was performed to identify the miRNA profile in the RIF-EVs. Participants/materials, setting, methods RIF-EVs and FER-EVs were characterized using western blotting, nanoparticle tracking analysis, and transmission electron microscopy. After co-culture with EVs, embryonic blastocyst rate and hatching rate were calculated. Besides, the proliferation, migration, and invasion of EV-treated trophoblast cells were evaluated by CCK–8, wound healing, and transwell invasion assays. miRNA expression profiles were compared between RIF-EVs and FER-EVs, and the regulatory role of significantly upregulated miR–6131 in RIF-EVs was investigated in the trophoblast cells. Main results and the role of chance RIF-EVs and FER-EVs are round bilayer vesicles, ranging mainly at 100 nm and enriched in TSG101, Alix, and CD9. Both RIF-EVs and FER-EVs entered embryonic or trophoblast cytoplasm. The blastocyst rate in the RIF-EV groups was significantly decreased compared to that in the FER-EV groups, at concentrations of 5, 10, and 20 μg/ml. The hatching rate was decreased significantly in embryos treated with 10 or 20 μg/ml RIF-EVs compared to those treated with FER-EVs at the same concentration (p < 0.05). The proliferation, migration, and invasion of trophoblasts were significantly decreased in the RIF-EV group at 20 μg/mL. A total of 11 differently expressed (fold change >2 and p < 0.05) miRNAs were found in the RIF-EVs, and two of them were validated in a larger set of EV samples using RT-PCR. The most significantly different miRNA, 6131, was increased in the RIF-EV-treated HTR8/SVneo cells. The up-regulation of miR–6131 inhibited the growth and invasion of HTR8/SVneo. Bioinformatics coupled with luciferase and western blot assays revealed that PAK2 is a direct target of miR–6131, and the overexpression of PAK2 can rescue the phenotype changes induced by miR–6131 overexpression. Limitations, reasons for caution Our study indicated miRNA in the RIF-EVs dysregulating the growth and function of embryonic cells. However, EVs contained a wide spectrum of bioactive molecules, including proteins, mRNAs, and DNA, which may play an important role in the implantation. Further studies are required to investigate the mechanisms. Wider implications of the findings: This work indicates an important role of EVs from women with RIF in embryonic implantation, potentially providing a novel insight to understand the pathophysiology of RIF. Trial registration number Not applicable

P-388 Pre-selected for an award: Endometrial extracellular vesicles from recurrent implantation failure patients inhibited embryonic growth and implantation via miR-6131/PAK2 pathway

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Liu
Keyword(s):  
Extracellular Vesicles ◽  
Migration And Invasion ◽  
Hatching Rate ◽  
Implantation Failure ◽  
Trophoblast Cells ◽  
Recurrent Implantation Failure ◽  
Endometrial Cells ◽  
Blastocyst Rate ◽  
Fertile Women

Abstract Study question Could endometrial extracellular vesicles from recurrent implantation failure patients (RIF-EVs) attenuate the growth and implantation potentials of embryos and what are the mechanisms?  Summary answer RIF-EVs inhibited embryonic growth and decreased the trophoblast functions via miR-6131/PAK2 pathway. What is known already Recurrent implantation failure (RIF) is characterized by repeated embryo transfers without pregnancy. To date, the etiology of RIF remains poorly understood. Recent evidence indicated that extracellular vesicles (EVs) secreted by endometrial cells, played a crucial role in the implantation by regulating the development and implantation of embryos. Study design, size, duration Endometrial cells isolated from endometrial tissues of RIF patients (n = 25) and fertile women (n = 16) were cultured and modulated via hormones. Endometrial EVs from RIF patients (RIF-EVs) or fertile women (FER-EVs) were isolated from the conditioned medium. The influence of EVs on embryonic development and implantation was investigated by co-culture models of EVs and 2-cell murine embryos or HTR8/SVneo cells, respectively. High-through put sequencing was performed to identify the miRNA profile in the RIF-EVs. Participants/materials, setting, methods RIF-EVs and FER-EVs were characterized using western blotting, nanoparticle tracking analysis, and transmission electron microscopy. After co-culture with EVs, embryonic blastocyst rate and hatching rate were calculated. Besides, the proliferation, migration, and invasion of EV-treated trophoblast cells were evaluated by CCK-8, wound healing, and transwell invasion assays. miRNA expression profiles were compared between RIF-EVs and FER-EVs, and the regulatory role of significantly upregulated miR-6131 in RIF-EVs was investigated in the trophoblast cells. Main results and the role of chance RIF-EVs and FER-EVs are round bilayer vesicles, ranging mainly at 100 nm and enriched in TSG101, Alix, and CD9. Both RIF-EVs and FER-EVs entered embryonic or trophoblast cytoplasm. The blastocyst rate in the RIF-EV groups was significantly decreased compared to that in the FER-EV groups, at concentrations of 5, 10, and 20 μg/ml. The hatching rate was decreased significantly in embryos treated with 10 or 20 μg/ml RIF-EVs compared to those treated with FER-EVs at the same concentration (p < 0.05). The proliferation, migration, and invasion of trophoblasts were significantly decreased in the RIF-EV group at 20 μg/mL. A total of 11 differently expressed (fold change >2 and p < 0.05) miRNAs were found in the RIF-EVs, and two of them were validated in a larger set of EV samples using RT-PCR. The most significantly different miRNA, 6131, was increased in the RIF-EV-treated HTR8/SVneo cells. The up-regulation of miR-6131 inhibited the growth and invasion of HTR8/SVneo. Bioinformatics coupled with luciferase and western blot assays revealed that PAK2 is a direct target of miR-6131, and the overexpression of PAK2 can rescue the phenotype changes induced by miR-6131 overexpression. Limitations, reasons for caution Our study indicated miRNA in the RIF-EVs dysregulating the growth and function of embryonic cells. However, EVs contained a wide spectrum of bioactive molecules, including proteins, mRNAs, and DNA, which may play an important role in the implantation. Further studies are required to investigate the mechanisms. Wider implications of the findings This work indicates an important role of EVs from women with RIF in embryonic implantation, potentially providing a novel insight to understand the pathophysiology of RIF. Trial registration number not applicable

scholarly journals The Expression of miR-31 and its Target Gene FOXP3 in Recurrent Implantation Failure Patients

2020 ◽  
Vol 8 (4) ◽  
pp. 389-395
Author(s):  
Azita Azarpoor ◽  
Abdolreza Ardeshirylajimi ◽  
Samira Mohammadi Yeganeh ◽  
Elham Pour matrood ◽  
Zeinab Dehghan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Target Gene ◽  
Control Group ◽  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
Tissue Samples ◽  
Significant Difference ◽  
Immunological Disorder ◽  
Recurrent Abortions ◽  
Fertile Women

Objectives: Endometrial receptivity is a complex event that occurs during the midluteal phase of the menstrual cycle known as the "window of implantation". During this period, the endometrium develops characteristics that allow the adhesion and invasion of the embryo to the uterine epithelium. Accordingly, the expressions of miR-31 and its target gene were evaluated to study the effect of miR-31 on FOPX3 gene expression in recurrent implantation failure (RIF) patients and normal fertile women. More precisely, the aim of this study was to understand the expression of miR-31 as one of the important regulators of the FOXP3 gene in the endometrium of RIF patients versus receptive endometria from fertile patients. Materials and Methods: This case-control study was conducted on 20 endometrial tissue samples of normal fertile women and RIF patients in order to evaluate miR-31 and its target gene expression. Results: According to the results of this study, a significant difference existed between RIF patients and normal fertile women (control group). The expression of the FOXP3 gene was more significant in the control group. miR-31 was also significantly expressed, which was due to the endometrial immunological disorder leading to the decreased expression of its target gene (FOXP3). Conclusions: In general, implant abnormalities and recurrent abortions were observed in RIF patients due to the decreased expression of the FOXP3 gene resulting from the inhibitory effects of miR-31.

scholarly journals ERAP, KIR, and HLA-C Profile in Recurrent Implantation Failure

2021 ◽  
Vol 12 ◽  
Author(s):  
Karolina Piekarska ◽  
Paweł Radwan ◽  
Agnieszka Tarnowska ◽  
Andrzej Wiśniewski ◽  
Michał Radwan ◽  
...  
Keyword(s):  
Nk Cells ◽  
Killer Cell ◽  
Hla Class I ◽  
Class I ◽  
Extravillous Trophoblast ◽  
Control Group ◽  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
Fertile Women

The mother’s uterine immune system is dominated by uterine natural killer (NK) cells during the first trimester of pregnancy. These cells express killer cell immunoglobulin-like receptors (KIRs) of inhibitory or activating function. Invading extravillous trophoblast cells express HLA-C molecules, and both maternal and paternal HLA-C allotypes are presented to KIRs. Endoplasmic reticulum aminopeptidase 1 (ERAP1) and 2 (ERAP2) shape the HLA class I immunopeptidome. The ERAPs remove N-terminal residues from antigenic precursor peptides and generate optimal-length peptides to fit into the HLA class I groove. The inability to form the correct HLA class I complexes with the appropriate peptides may result in a lack of immune response by NK cells. The aim of this study was to investigate the role of ERAP1 and ERAP2 polymorphisms in the context of KIR and HLA-C genes in recurrent implantation failure (RIF). In addition, for the first time, we showed the results of ERAP1 and ERAP2 secretion into the peripheral blood of patients and fertile women. We tested a total of 881 women. Four hundred ninety-six females were patients who, together with their partners, participated in in vitro fertilization (IVF). A group of 385 fertile women constituted the control group. Women positive for KIR genes in the Tel AA region and HLA-C2C2 were more prevalent in the RIF group than in fertile women (p/pcorr. = 0.004/0.012, OR = 2.321). Of the ERAP polymorphisms studied, two of them (rs26653 and rs26618) appear to affect RIF susceptibility in HLA-C2-positive patients. Moreover, fertile women who gave birth in the past secreted significantly more ERAP1 than IVF women and control pregnant women (p < 0.0001 and p = 0.0005, respectively). In the case of ERAP2, the opposite result was observed; i.e., fertile women secreted far less ERAP2 than IVF patients (p = 0.0098). Patients who became pregnant after in vitro fertilization embryo transfer (IVF-ET) released far less ERAP2 than patients who miscarried (p = 0.0032). Receiver operating characteristic (ROC) analyses indicate a value of about 2.9 ng/ml of ERAP2 as a point of differentiation between patients who miscarried and those who gave birth to a healthy child. Our study indicates that both ERAP1 and ERAP2 may be involved in processes related to reproduction.

scholarly journals Alterations of Uterine Blood Flow During the Follicular Phase in Patients With Recurrent Implantation Failure: A Doppler Ultrasonographic Study

2021 ◽  
Vol 9 (3) ◽  
pp. 217-221
Author(s):  
Marjan Amini ◽  
Mahnaz Ranjkesh ◽  
Saba Nikanfar ◽  
Amir Fattahi ◽  
Laya Farzadi ◽  
...  
Keyword(s):  
Blood Flow ◽  
Color Doppler ◽  
Follicular Phase ◽  
Control Group ◽  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
History Of ◽  
Uterine Perfusion ◽  
Endometrial Blood Flow ◽  
Fertile Women

Objectives: The dynamics of blood flow in the endometrium plays a crucial role during the implantation process. This study aimed to assess the uterine perfusion during the follicular phase in patients with a history of recurrent implantation failure (RIF) and healthy fertile women using the transvaginal ultrasound color Doppler method. Materials and Methods: To this end, 50 patients with RIF and 50 age-matched healthy fertile women were recruited in this case-control study. The transvaginal color Doppler ultrasonography was used to evaluate the pulsatility index (PI) and resistance index (RI) of the uterine, arcuate, and sub-endometrial arteries during the follicular phase in both groups. Results: The RI and PI of both right and left uterine arteries were higher in the RIF group compared to the fertile women (P<0.05). Our results showed that the PI and RI of sub-endometrial blood flow and the RI of arcuate arteries were substantially higher in the group with a history of RIF in comparison with the control group. However, the PI of arcuate arteries was not significantly different between the groups. Conclusions: Adequate uterine perfusion and sub-endometrial blood flow are necessary to achieve successful implantation and pregnancy since our results demonstrated the higher resistance of uterine and sub-endometrial arteries in patients with a history of RIF. Thus, the assessments of uterine perfusion indices during the follicular phase could be used as a non-invasive method in the evaluation of patients with RIF.

scholarly journals Uterine and Serum Glycodelin Concentration in Recurrent Implantation Failure Versus Normal Fertile Women on Implantation Window

2017 ◽  
Vol 5 (2) ◽  
pp. 103-106 ◽  
Author(s):  
Robabeh Taheripanah ◽  
Marzieh Zamaniyan ◽  
Shahrzad Akhoondzadeh ◽  
Anahita Taheripanah ◽  
Narges Malih
Keyword(s):  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
Implantation Window ◽  
Fertile Women
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