Repeated Implantation Failure Patients Research Articles

Personalized embryo transfer guided by rsERT improves pregnancy outcomes in patients with repeated implantation failure

IntroductionEmbryo implantation requires synchronous communication between the embryo and maternal endometrium. Inadequate maternal endometrial receptivity is one of the principal causes for embryo implantation failure [especially repeated implantation failure (RIF)] when biopsied good-quality euploid embryos are transferred. An RNA-seq-based endometrial receptivity test (rsERT) was previously established to precisely guide successful embryo implantation. In this study, we aimed to evaluate the effect of personalized embryo transfer (pET) via rsERT on the clinical outcomes in patients with RIF.MethodsA total of 155 patients with RIF were included in the present retrospective study and were divided into two groups: 60 patients who underwent rsERT and pET (Group rsERT) and 95 patients who underwent standard frozen embryo transfer (FET) without rsERT (Group FET). Reproductive outcomes were compared for patients who underwent rsERT-guided pET and standard FET.ResultsForty percent (24/60) of the patients who underwent rsERT were receptive, and the remaining 60% (36/60) were non-receptive. The positive human chorionic gonadotropin (β-hCG) rate (56.3% vs. 30.5%, P = 0.003) and clinical pregnancy rate (43.8% vs. 24.2%, P = 0.017) were significantly higher in Group rsERT patients than in FET group patients. Additionally, Group rsERT patients also showed a higher implantation rate (32.1% vs. 22.1%, P = 0.104) and live birth rate (35.4% vs. 21.1%, P = 0.064) when compared with FET patients, although without significance. For subpopulation analysis, the positive β-hCG rate, clinical pregnancy rate, implantation rate, and live birth rate of receptive patients were not statistically significant different from those of non-receptive patients.ConclusionsThe rsERT can significantly improve the pregnancy outcomes of RIF patients, indicating the clinical potential of rsERT-guided pET.

Metabolomic profiling of embryo culture media in patients with repeated implantation failure during assisted reproductive technology cycles.

This study investigated the metabolic status of the spent culture media from embryos of patients with repeated implantation failure (RIF) undergoing in vitro fertilization-intracytoplasmic sperm injection cycles in comparison with the embryos from healthy fertile women. Metabolite levels in spent culture media were assessed and compared between embryos from RIF patients (n=35) and oocyte donors as controls (n=15). Protein levels of insulin-like growth factor 1 (IGF-1) were determined using Western blotting. Concentrations of glucose, pyruvate, and lactate were measured using spectrophotometry. Ionic colorimetric assay kits were utilized to analyze the concentrations of sodium, chloride, calcium, and magnesium ions. High-performance liquid chromatography was employed to measure the concentrations of glutamic acid, aspartic acid, methionine, phenylalanine, and histidine. Glucose consumption and lactate secretion were higher in the control group than in the RIF group. The magnesium concentration was significantly higher in the control group than in the RIF group, but glutamic acid and aspartic acid concentrations were lower in the control group than in the RIF patients (p<0.05). The levels of IGF-1, sodium, calcium, chloride, methionine, histidine, and phenylalanine did not show statistically significant differences between the two groups. The metabolic profile of the culture medium of the embryos in the RIF group differed from that of the control group. These findings suggest potential factors that may affect implantation capacity in RIF patients and provide a new perspective on embryo selection.

#238 : rsERT Guided Personalized Embryo Transfer Improves Clinical Outcomes in RIF Patients

Background and Aims: Narrowed or shifted window of implantation (WOI) is acknowledged to be one of the endometrial origins of embryo implantation failure, especially in patients with repeated implantation failure (RIF). RNA-Seq-based endometrial receptivity test (rsERT) was developed to assess the endometrial receptivity (ER) and guide the personalized embryo transfer (pET). Clinical studies were conducted to evaluate the clinical utility of rsERT in RIF patients. Method: 566 RIF patients with rsERT and pET in multi-centers from 2019-2022 were included in this retrospective study, which was defined as rsERT-pET group. RIF patients without rsERT and pET in the same period was used as control for comparison. rsERT-pET group was further divided into normal WOI (nWOI) and shifted WOI (sWOI) groups. Clinical pregnancy rate and ongoing pregnancy rate were therefore compared among these groups by nonparametric statistical approaches. Results: The intrauterine pregnancy and ongoing pregnancy rates of rsERT-pET group were receptively 55.0% and 46.2%, which were significantly higher than those of the control group (55.0% vs 35.1%, OR = 2.26, p-value = 2.33×10[Formula: see text]; 46.2% vs 19.8%, OR = 3.45, p-value = 4.08×10[Formula: see text] ). We further evaluated the intrauterine pregnancy rates of sWOI and nWOI and found remarkably better pregnancy outcomes in patients from nWOI group compared to those in sWOI group patients (62.4% vs 50.4%, OR = 1.64, p-value = 0.005). Conclusion: The pET guided by the result of rsERT significantly improved the clinical outcomes in patients with RIF based on the intrauterine pregnancy rate and the ongoing pregnancy rate, indicating the clinical potential of rsERT method.

#199 : Does Hysteroscopic Evaluation of Uterine Characteristic Improve Outcome of Thin Endometrium in Patients with a History of Repeated Implantation Failures?

Background: Repeated implantation failure (RIF) is a problem for many patients suffering from infertility and their physicians. Despite numerous developments in the field of assisted reproduction, the implantation rate remains low. Among the various reasons of implantation failure, endometrial regularity has an important role. Recently, hysteroscopy is an accurate method for evaluating the endometrial integrity, with the ability to treat uterine pathology for repeated implantation failure patients. Objective: The aim of this study to evaluate the incidence of uterine pathology among infertile women with a history of two implantation failures after IVF and estimate the effect of diagnostic hysteroscopy and hysteroscopic procedures in uterine cavity on achieving a pregnancy in these patients in Mongolia. Materials and Methods: The retrospective study population included infertile women who attended the outpatient department of the Creation and Love Women’s infertility clinic, between August 2020 and July 2022 in Mongolia. The study inclusion criteria were women who had more than two RIF of IVF/ET cycle. Pregnancy and nonpregnancy rate after hysteroscopic procedures were analyzed included in patients reports archived in clinical data. Results: Hysteroscopy were performed on 35 women who had at least two RIFs. The participants’ mean age was 40.3±5.3 and mean duration of infertility year was evaluated 5.53±3.7. Mean AMH level was 1.4±1.6 ng/ml. All patients were diagnosed abnormal uterine cavity by hysteroscopy. Out of 35 patients participated in this study who underwent hysteroscopic evaluation, 20 patients (57.1%) showed synechiae (mild, moderate and severe synechiae). Second most frequent hysteroscopic finding was the endometrial polyp, accounting for 15 (42.9%) patients. Pregnancy rate after hysteroscopic procedures was 8 patients (23%) in this study group. The duration of infertility year was significantly different in pregnant and nonpregnant women underwent hysteroscopic correction (p&lt;0.05). Conclusion: The intrauterine pathology diagnosed by hysteroscopy in patients with previous IVF failure were found all patients in this study. The synechiolysis was common abnormality in Mongolia may cause by artificial abortion. The correction of abnormalities such as and polyp synechiolysis might improve the outcome of successful pregnancy rate.

Upregulated RPA2 in endometrial tissues of repeated implantation failure patients impairs the endometrial decidualization.

To investigate the expression and underlying mechanism of RPA2 in endometrium of patients with repeated implantation failure (RIF). In this study, we retrieved the expression profiles from GEO databases and filtered the differentially expressed genes between RIF and the fertile control group. Ultimately, RPA2 was confirmed as a target gene. RPA2 expression in endometrial tissues of RIF patients, the control group, and different phases was detected by RT-qPCR, immunohistochemistry, and Western blotting. The role of RPA2 in endometrial decidualization was performed by in vitro decidualization inducing by 8-Br-cAMP and MPA. Furthermore, RT-qPCR was used to detect changes in the decidual biomarkers after transfection of RPA2 overexpression vector in human endometrium stromal cell (HESC). RPA2 was significantly upregulated in the mid-secretory endometrium of patients with RIF. As a proliferation-related gene, RPA2 was obviously higher expressed at proliferative phase during the normal menstrual cycles. Moreover, the downregulation of RPA2 was discovered during decidualization of HESC. Furthermore, RPA2 overexpression impaired decidualization by inhibiting the expression of prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1). Our finding indicated that aberrant upregulation of RPA2 attenuated decidualization of HESC in RIF women and provided new potential therapeutic targets.

Efficacy of the long-acting gonadotropin-releasing hormone agonist long protocol on IVF/ICSI outcomes of patients with repeated implantation failure.

The present study aimed to investigate the effect of long-acting gonadotropin-releasing hormone agonist (GnRHa) long protocol on in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcomes of patients with repeated implantation failure (RIF). The present study was carried out from June 1, 2016 to June 30, 2021. A total of 665 patients with RIF were enrolled into the study and classified by the ovarian stimulation protocols. The outcome parameters were compared in each group. In addition, we evaluated the expression of homeobox A10 (HOXA10), integrin β3 and leukemia inhibitory factor (LIF) in endometrial tissues between groups by quantitative RT-PCR. Patients who received the long-acting GnRHa long protocol had significantly higher clinical pregnancy rates (58.0%, 41.7% and 39.9%, respectively; P = 0.008 and 0.003), implantation rates (38.1%, 30.3%, and 30.1%, respectively; P = 0.001 and <0.001) and live birth rates (50.3%, 36.3%, and 31.3%, respectively; P = 0.020 and 0.002) compared with the short-acting GnRHa long protocol and GnRH antagonist protocol. In addition, we found that long-acting GnRHa could improve the expression of HOXA10 (P < 0.05). The long-acting GnRHa long protocol could improve endometrial receptivity and IVF/ICSI clinical outcomes of RIF patients compared with the short-acting GnRHa long protocol and GnRH antagonist protocol.

Effect of Chinese patent medicine Kunling Pill on endometrial receptivity: A clinical trial, network pharmacology, and animal-based study.

Although pregnancy success rates are raised with assisted reproductive technology, it still cannot meet clinical demands. Kunling Pill (KLP), a traditional Chinese medicine, is widely used in various gynecological disorders, particularly in improving fertility and pregnancy rates. However, the underlying mechanism of how KLP affects pregnancy outcomes remains unclear. This study aimed to explore the effects and mechanisms of KLP on endometrial receptivity. Firstly, a retrospective trial was conducted to validate the efficacy of KLP on repeated implantation failure (RIF) patients. The result indicated a significant increase in the proportion of live birth in KLP group (30.56%) compared to the control group (16.89%). Secondly, network pharmacology methods predicted the active components and network targets of KLP. Endometrial receptivity is closely associated with the activation of inflammatory factors, predicting the function of KLP on the immune system. The estrogen and apoptotic signaling pathways were also highlighted in the gene ontology enrichment analysis. Thirdly, a decreased endometrial receptivity model was established by controlled ovarian hyperstimulation (COH) in female C57BL/6 mice, divided into the COH and KLP groups. Normal female mice are as control group. In vivo, KLP administration could increase endometrial thickness and the number of endometrial glands and pinopodes. In the endometrium, KLP supplementation upregulated the expressions of estrogen receptor α, progesterone receptor, endothelial nitric oxide synthase, and integrin αVβ3 in the murine uterus and reduced serum levels of estrogen and progesterone. KLP regulated the uterine immune cells and inhibited cell apoptosis in the ovary via Bcl-2/Bax/caspase-3 pathway. In conclusion, KLP administration raised the live birth rate in RIF patients to optimize medication regimens, mainly because KLP ameliorated impaired endometrial receptivity.

Circulating and Endometrial Profiles of miR-145, miR-155-5p, miR-224, MPP-5, and PECAM-1 Expression in Patients with Repeated Implantation Failure: A Case Control Study.

An association between microRNAs (miRNAs) and adhesion proteins expression with repeated implantation failure (RIF) has been recently reported; however, these findings are controversial. This study aims to evaluate the endometrial and circulating expressions of miR-145, miR-155-5p, and miR-224 in addition to the endometrial expressions of membrane protein palmitoylated-5 (MPP-5) and endothelial cell adhesion molecule-1 (PECAM-1) in patients with RIF compared to control subjects. This case-control study was carried out between June 2021-July 2022. Subjects included 17 patients with RIF and 17 control subjects, who had previous spontaneous term pregnancy with a live birth, who referred to the Medical Centre of Arash Hospital, Tehran, Iran. Endometrial tissue samples were obtained via hysteroscopy and Pipelle catheter in the RIF and control subjects, respectively. Plasma samples were collected after ovulation in all subjects. The expression levels of MPP5, PECAM-1, miR-224, miR-145, and miR-155-5p were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). The student's t test, chi-square, Mann-Whitney U, and analysis of covariance (ANCOVA) were used for data analyses. RIF patients had less endometrial miR-155-5p expression, and higher endometrial and circulating expressions of miR-145 and miR-224 compared to control subjects. Endometrial PECAM-1 and MPP5 expression significantly decreased in patients with RIF compared to the control group. There was a positive correlation between circulating miR-224 and endometrial miR-155-5p, and between circulating miR-155-5p and endometrial PECAM-1 expression levels in patients with RIF. The present study suggests that circulating miR-224, endometrial miR-145, and PECAM-1 can be reliable, novel biomarkers for diagnosis of RIF.

P-485 Performance of ERA test after PGT-A in RIF patients defined according to ESHRE 2023 good practice recommendations: a retrospective case-control study

Abstract Study question Is Endometrial-Receptivity-Array (ERA) test clinically-useful among Repeated-Implantation-Failure (RIF) women defined according to ESHRE 2023 good-practice-recommendations (GPR) and undergoing single euploid blastocyst transfer? Summary answer ERA-test showed a higher “non-receptive endometrium” rate among RIF. Nevertheless, the live-birth-rate (LBR) per first euploid transfer was independent from progesterone timing according to ERA-test. What is known already ERA-test evaluates the expression of 238 genes from an endometrial biopsy. It aims at predicting the window of implantation and adjust progesterone timing accordingly. The data are controversial due to poor study design and/or limited sample size. Few studies investigated ERA-test in a setting with euploid transfers and/or among RIF patients. Moreover, RIF definition is largely inconsistent leading to a significant bias in the interpretation of the existing literature and treatment. ESHRE 2023 GPR revised RIF definition. Here, we leveraged this updated definition and PGT-A to assess ERA-test diagnostic/therapeutic performance in these women. Study design, size, duration Retrospective case-control study during 2013-2021 including 2211 patients with at least one euploid blastocyst after PGT-A. Hysteroscopy was conducted in case of suspect uterine pathologies, biopsy in case of suspect inflammation, and therapy in case of endometritis. 265 patients (12%) were classified RIF according to the ESHRE GPR (= no successful implantation despite an estimated cumulative chance ≥60%). 48 RIF patients requested ERA-test before undergoing vitrified-warmed euploid single blastocyst transfer, while 217 did not. Participants/materials, setting, methods qPCR/NGS-based PGT-A was conducted to assess non-mosaic aneuploidies. Normal thyroid function, vitamin-D levels, uterine cavity, and absence of sactosalpynx were confirmed. Whenever the ERA test suggested altered receptivity, progesterone timing was modified accordingly. The primary outcome was LBR per first euploid transfer in the non-ERA versus ERA groups, with a sub-analysis in ERA receptive versus non-receptive. Outcomes were adjusted for confounders assessed via logistic regressions. Main results and the role of chance RIF patients requesting ERA-test were older (39.1±4.5 versus 36.7±3.5 years, p &amp;lt; 0.01), and had already undergone 3.7±2.0 transfers resulted in implantation failures (1.5±1.3 of euploid blastocysts) versus 2.6±1.7 transfers (1.0±0.8 of which of euploid blastocysts) in the control (p &amp;lt; 0.01). Nevertheless, all these variables were not associated with the primary outcome under investigation. 44 non-RIF patients requested ERA-test in the same study period. Notably, the rate of pre-/post-receptive endometria was 39.6% (N = 19/48) among RIF and 13.6% (N = 6/44) among non-RIF women (p &amp;lt; 0.01). In fact, maternal age was not associated with “non-receptive” responses after ERA-test, the number of previous implantation failures showed an Odds-Ratio 1.29, 95%CI 1.03-1.62, p = 0.03. Blastocyst quality and day of full-blastulation were similar in ERA and non-ERA patients, as well as in ERA receptive and non-receptive patients. Positive pregnancy test, biochemical-pregnancy-loss, and miscarriage rates were all similar among non-ERA and ERA, and among ERA receptive and non-receptive patients. Indeed, also the LBR was similar among non-ERA and ERA (N = 98/220, 44.5% versus N = 17/48, 35.4%, p = 0.26; Odds-Ratio adjusted for blastocyst quality and day of full-blastulation: 0.76, 95%CI:0.39.1.48, p = 0.42) and among ERA receptive and non-receptive patients (N = 10/29, 34.5% versus N = 7/19, 36.8%, p = 0.99; Odds-Ratio adjusted for blastocyst quality and day of full-blastulation: 0.99, 95%CI:0.3-3.4, p = 0.99). Limitations, reasons for caution Our standard clinical workflow does not entail ERA-test, but the couple may request it. The study is retrospective with limited sample size. Patients requesting ERA-test were older with more previous failures. Non-ERA and ERA receptive patients underwent either Hormone-Replacement-Therapy or Modified-Natural-Cycle. Cost-effectiveness analyses are missing. Wider implications of the findings A promising association was reported between the novel definition of RIF and “non-receptive” responses, suggesting that ERA-test might unveil endometrial receptivity issues. Nevertheless, positive, and negative predictive values upon implantation are largely missing and its clinical utility as a therapeutic tool in RIF women remains questionable. Trial registration number none

Mitochondrial energy metabolism is downregulated in repeated implantation failure patients related to alteration of PGC-1α acetylation level.

Herein we aimed at exploring mitochondrial energy metabolism status in patients with repeated implantation failure (RIF) and whether key regulatory factor PGC-1α of energy metabolism is involved in the decidualization of endometrial stromal cells. Mitochondrial oxidative phosphorylation level and ATP synthesis were compared in primary endometrial stromal cells from RIF and control group. At the same time, as one of key transcription regulators of mitochondrial energy metabolism, the expression level and acetylation level of PGC-1α were compared with two groups. Then, we downregulated the acetylation levels of PGC-1α, and the expression of decidual markers (PRL and IGFBP1) was observed further. Mitochondrial energy metabolism, showing by mitochondrial oxidative phosphorylation level and ATP synthesis, was decreased in the endometrial stromal cells of the RIF group (RIF-hEnSCs). Meanwhile, PGC-1α acetylation levels were significantly higher in RIF-hEnSCs. When we reduced the acetylation levels of PGC-1α in RIF-hEnSCs, the basal O2 consumption rate and maximal respiration were increased, and also the PRL and IGFBP1. Overall, our data indicated that the endometrial stromal cells of the RIF patients had low level of mitochondrial energy metabolism. Reducing acetylation level of key energy metabolism regulator PGC-1α can increase the decidualization level of RIF-hEnSCs. These findings may inspire new ideas about the treatment of RIF.

Histological endometrial dating: a reliable tool for personalized frozen-thawed embryo transfer in patients with repeated implantation failure in natural cycles

ObjectiveTo evaluate the clinical availability and stability of histological endometrial dating as a tool for personalized frozen-thawed embryo transfer (pFET) in patients with repeated implantation failure (RIF) in natural cycles.MethodsA total of 1245 RIF patients were recruited to the present study. All of the patients received an endometrial dating evaluation on day 7 post-ovulation (PO + 7) to guide their first pFET. The second and third pFETs were executed according to histological examination (again employing biopsy) or by reference to previous results. Subsequent pregnancy outcomes for all of the cycles were ultimately tracked.ResultsThe out-of-phase rate for RIF patients was 32.4% (404/1245) and the expected dating rate (the probability of the expected endometrial dating aligning with repeat biopsy) for endometrial dating reevaluation was as high as 94.3% (50/53). The clinical pregnancy rates of first, second, and third pFETs were 65.3%, 50.0%, and 44.4%, respectively; and the cumulative clinical pregnancy rate attained 74.9% after three transfers. Endometrial dating reevaluations met expectations with more than a 2-year duration in three cases and elicited favorable clinical outcomes.ConclusionWe validated the relatively high stability of the histological endometrial dating platform—including the out-of-phase rate and the expected dating rate of reevaluation in patients with RIF—by expanding the sample size. The pFET, based on histological endometrial dating, was of acceptable clinical value and was worthy of promotion in patients with unexplained RIF.

Impact of body mass index on peripheral and uterine immune status in the window of implantation in patients with recurrent reproductive failure

This study aimed to investigate whether maternal obesity affects the immune status of peripheral blood and endometrium in patients with recurrent reproductive failure classified according to their body mass index (BMI). A total of 228 repeated implantation failure (RIF) and 266 recurrent miscarriage (RM) patients were enrolled in the study and further subdivided into three groups according to their BMI: (i) normal weight (18.5≤ BMI <23); (ii) overweight (23≤ BMI <25); and (iii) obese (BMI ≥25). Peripheral blood and endometrium samples were collected in the mid-luteal phase before IVF treatment or natural pregnancy. Peripheral immunocytes were analyzed by flow cytometry, while uterine immune cells were subjected to immunohistochemistry. In RM patients, significantly increased peripheral helper T cells and decreased cytotoxic T cells, NK cells were observed in the obese group compared with the normal-weight group. Meanwhile, in the endometrium, the percentage of NK cell, macrophage cell, M2 macrophage cell, and Treg cell significantly reduced with increased BMI in RIF patients, and the percentage of NK cell and M2 macrophage cell significantly decreased with increased BMI in RM patients. In conclusion, obesity may cause endometrial immune disorder in recurrent reproductive failure women, but was only associated with the peripheral immune change in RM patients.

Personalized embryo transfer based on RNA sequencing endometrial receptivity test in repeated implantation failure patients: artificial cycle versus natural cycle

Introduction Embryo implantation provides an efficient way for patients with repeated implantation failure (RIF) to achieve pregnancy. The aim of this study is to compare the implantation outcomes of RIF patients in artificial cycle to those in natural cycle, both were treated with RNA sequencing endometrial receptivity test (rsERT) based personalized embryo implantation. Methods The endometrial receptivity (ER) analysis was performed using rsERT followed by personalized embryo transfer at optimal window of implantation (WOI). The implantation rate (IR), clinical pregnancy rate (CPR) and live birth rate (LBR) were calculated. The expression levels of biomarkers involved in pregnancy process in the patients detected as in receptivity status were also analyzed. Results The rsERT shown that 44.8% (natural cycle) and 47.8% (artificial cycle) patients were in non-receptive status, which indicated a WOI displacement. After personalized embryo transfer, the IR of patients in artificial cycle was higher than those in natural cycle (52.2% vs 27.6%). The expressions of FKBP52, MUC1 and LPAR3 were significantly lower in artificial cycle than in natural cycle. Conclusion Using artificial cycle for personalized embryo transfer based on rsERT may yield better pregnancy outcomes for RIF patients. A gene expression analysis of FKBP52, MUC1 and LPAR3 provided a potential way to increase implantation outcomes for RIF patients.

ITRAQ-based Proteomic Analysis Unveils ACSL4 as a Novel Potential Regulator of Human Endometrial Receptivity.

Competent endometrial receptivity is a prerequisite for successful embryo implantation. Identification of novel key molecules involved in endometrial receptivity is essential to better interpret human implantation and improve pregnancy rates in assisted reproduction treatment. Isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomics was performed to profile the proteomes of the prereceptive (luteinizing hormone [LH] + 2, n = 4) and receptive (LH + 7, n = 4) endometrial tissues. A total of 173 differentially expressed proteins (DEPs) between LH + 2 and LH + 7 endometrial samples were identified. Integrated analysis of the proteomic data and published transcriptomic data was performed to identify the concordant DEPs with differential expression at both the messenger RNA and protein levels. Protein-protein interaction (PPI) network analysis was performed on concordant DEPs. We first identified 63 novel concordant DEPs and 5 hub proteins (ACSL4, ACSL5, COL1A1, PTGS1, and PLA2G4F) between LH + 2 and LH + 7 endometrial samples. ACSL4 was predominantly expressed in endometrial epithelial cells and its expression was significantly upregulated by progesterone in the LH + 7 endometrium and significantly downregulated in repeated implantation failure patients. Knockdown of ACSL4 in endometrial epithelial cells induced the downregulation of endometrial receptivity markers (HOXA10, COX2, and LIF) and the significant decrease of implantation rate during in vitro implantation analysis. This study provides the first gel-independent quantitative proteomes of the LH + 2 and LH + 7 human endometrium using iTRAQ technology. The identified concordant DEPs and hub proteins open a new avenue for future studies aimed at elucidating the underlying mechanisms governing endometrial receptivity. ACSL4 was identified as a novel regulatory molecule in the establishment of endometrial receptivity and might play important roles during implantation.

Study of LIF, NR3C1, BCL6, miR-494, and miR-30d in Endometrial Epithelial Cells of Repeated Implantation Failure and Healthy Women Treated with Calcitriol and Letrozole in a Fibrin Scaffold

Background: Determining the exact time of window of implantation (WOI) in individuals with repeated implantation failure (RIF) is necessary for successful implantation. The present study aimed to compare the effects of letrozole and/or calcitriol on the expression of receptivity markers in endometrial epithelial cells (EECs) of RIF patients and healthy fertile women. Methods: Endometrial tissue was obtained from RIF patients (n=5) and healthy women (n=5) in the proliferation phase. EECs were isolated and cultured in a fibrin scaffold in 10 groups. The cells were treated with letrozole and/or calcitriol in three types of treatment. To check the proliferation and migration of cells in the scaffold, immunofluorescent and H&amp;E staining was used, and gene expression was checked using real-time polymerase chain reaction (PCR). Results: This study showed that the expression levels of pro-implantation genes (NR3C1, LIF, and miR-30d) were higher in healthy women than in RIF patients, and the expression levels of anti-implantation genes (BCL6 and miR-494) were lower in healthy women than in RIF patients. Calcitriol and letrozole increased the expression level of pro-implantation genes (NR3C1, LIF, and miR-30d) in RIF patients. Furthermore, they reduced the expression level of anti-implantation genes (BCL6 and miR-494) in RIF patients. Conclusion: Our findings suggest that treatment with letrozole is more effective than co-treatment with letrozole and calcitriol or alone treatment with calcitriol on receptivity markers in RIF patients.

Cutting-edge techniques provide insights regarding repeated implantation failure patients

Research questionCan time-lapse parameters and the transcriptional profile of cumulus cells be used to achieve a more stringent and non-invasive method of embryo assessment and to identify possible factors affecting the embryo's ability to implant in repeated implantation failure (RIF) patients? DesignA total of 190 embryos from 18 oocyte donors and 145 embryos from 15 RIF patients were evaluated based on time-lapse parameters. Three morphokinetic parameters including T5 (time to reach five cells), T3 (time to reach three cells) and CC2 (time to two to three cells) were recorded for all embryos. Embryos that had all three parameters in the normal range were graded as high quality and comparison between these parameters were compared in high-quality embryos between two groups. The transcriptional profile of cumulus cells related to high-quality embryos of both groups were analysed by RNA sequencing and compared. Finally, the possible relationship between differentially expressed genes and time-lapse parameters was examined. ResultsT5 was significantly lower in the RIF group than the donor group (P = 0.011). The cumulus cell transcriptome analysis showed 193 genes were down-regulated and 222 genes up-regulated. The mammalian target of rapamycin and the transforming growth factor beta pathways were significantly increased in the RIF group compared to the donor group (P = 0.007 and 0.01, respectively). Vitamin B12 and fatty acid beta-oxidation pathways were also significantly reduced in the RIF group compared to the donor group (P = 0.006 and 0.01, respectively). ConclusionsDifferences in the transcriptomic profiles of cumulus cells and some morphokinetic parameters may be one of the main factors contributing to unexplained RIF.

The endometrial microbiota profile influenced pregnancy outcomes in patients with repeated implantation failure: a retrospective study

ObjectivesTo investigate the association between the pregnancy outcomes of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) patients with repeated implantation failure (RIF) and their endometrial microbiota profiles. MethodsOne hundred and forty-one RIF patients were recruited in this retrospective study. Endometrial tissues were sampled using a disposable sterile endometrium sampler. Comprehensive next-generation sequencing techniques were used to detect the endometrial bacteria status, and the pregnancy outcomes were analyzed. ResultsEndometrial pathogenic bacteria were detected in 125 patients (88.70%, the pathogenic group) while no relevant pathogen was found in the remaining 16 (11.30%, the no-pathogen group). All the 125 patients received the treatment of oral antibiotics for 14 days. Clinical pregnancy rates and ongoing pregnancy rates were higher in the pathogenic group than in the no-pathogen group without statistical significance (50.40% vs. 37.50%, P＞0.05; 42.40% vs. 25%, P＞0.05). ConclusionIn the endometrium of most RIF patients existed pathogenic bacteria, among which Streptococcus, Staphylococcus, Neisseria, and Klebsiella were most frequently observed, and the sensitive antibiotic therapy might improve clinical outcomes of the RIF patients in subsequent embryo transfer cycles.

Chronic endometritis impairs embryo implantation in patients with repeated implantation failure: A retrospective study

ObjectiveTo assess whether chronic endometritis (CE) affects embryo implantation in patients with repeated implantation failure (RIF). Materials and methodsWe retrospectively analyzed 126 RIF patients who were never diagnosed with CE and received no prior antibiotic therapy. Endometrial specimens obtained by endometrial scratching during mid-luteal phase were immunostained by CD138, a hallmark plasmacyte marker, to identify CE. Pregnancy outcome in RIF patients who underwent IVF-ET frozen-embryo within transfers 6 months after endometrial scratching was compared between women with and without CE. ResultsThe prevalence of CE in patients with RIF was found to be 11.9% (15/126). Moreover, a significantly reduced clinical pregnancy rate was observed in RIF patients with CE (20% vs. 46.85%; p = 0.04). The live birth rate also exhibited a decreasing trend in RIF patients with CE, although there was no statistically significant difference (20% vs. 41.58%; p = 0.109). ConclusionsCE may be involved in the failure of embryo implantation and reduced clinical pregnancy outcome in patients with RIF.

The effectiveness of immunomodulatory therapies for patients with repeated implantation failure: a systematic review and network meta-analysis

This meta-analysis analyzed the clinical pregnancy outcomes of repeated implantation failure (RIF) patients treated with immunomodulatory therapies. Publications (published by August 16, 2021) were identified by searching the PubMed, Embase, and Web of Science databases. The quality of the studies was evaluated with the Cochrane bias risk assessment tool, and a network meta-analysis was performed with Stata 14.0. The outcomes were clinical pregnancy rate (CPR), live birth rate (LBR), and implantation rate (IR). The results of our network meta-analysis of 16 RCTs (including 2,008 participants) show that PBMCs, PRP, and SC-GCSF can significantly improve the CPR compared with LMWH (PBMCs: OR 2.15; 95% CI 1.21–3.83; PRP: OR 2.38; 95% CI 1.08–5.24; SC-GCSF: OR 2.46; 95% CI 1.05–5.72). The LBR of PRP was significantly higher than those of IU-GCSF (OR 3.81; 95% CI 1.22–11.86), LMWH (OR 4.38; 95% CI 1.50–12.90), and intralipid (OR 3.85; 95% CI 1.03–14.29), and the LBR of PBMCs was also significantly better than that of LMWH (OR 2.35; 95% CI 1.14–4.85). Furthermore, PRP treatment significantly improved the IR compared with LMWH treatment (OR 2.81; 95% CI 1.07–7.4). The limited evidence from existing RCTs suggests that PBMCs and PRP are the best therapeutic options for RIF patients. However, owing to the quantity limitation, more top-quality research is required to obtain additional high-level evidence.

Potential utility of a non-invasive menstrual blood immunophenotype analysis in reproductive medicine.

Purpose Can a comprehensive flow cytometry panel be used to assess immunophenotype profiles in menstrual blood of patients experiencing reproductive failure and age matched controls of proven fertility? Methods 58 recurrent pregnancy loss and repeated implantation failure patients, along with 15 age matched controls of proven fertility, had menstrual blood samples obtained within the first 24 hours of the onset of menstruation to non-invasively assess the local immunophenotype. Using a comprehensive multi-parameter flow panel the lymphocyte sub-populations were described and compared. Results Relative to well established peripheral blood immunophenotyping values, distinct lymphocyte population differences were noted between the subgroups. The ratios of CD4+ and CD8+ T-cells were inverted relative to peripheral blood and uterine NK cells represented by CD56bright were distinctly visualised, emphasising the distinction of menstrual and peripheral blood. Relative to controls there were marked increases in CD3+ve T-cells (p=0.009), CD4:CD8 ratio (p=0.004), CD19 B-cells (p=0.026) and CD56dim NK's (p=0.002) in the reproductive failure cases. Conclusions Flow cytometric evaluation can provide a rapid and objective analysis of lymphocyte subpopulations in many forms of tissue and fluid. The findings show significant variations in cellular composition of immune cells indicating a distinct compartment, with differences between cases and controls. Immunological assessment of the menstrual blood immunophenotype, in clinically appropriate patients, may provide insight into the aetiology of adverse reproductive outcome, without the risks and inconveniences associated with a more invasive endometrial biopsy.