Artificial Neural Network Model Using Immune-infiltration Modules for Endometrial Receptivity Assessment of Implantation Failure

Objectives: This study was anchored on the state of local immune-infiltration in the endometrium, which acts as critical factors affecting embryonic implantation, and aimed at establishing novel approaches to assess endometrial receptivity for patients with IVF failure.Methods: Immune-infiltration levels in the GSE58144 dataset (n=115) from GEO were analyzed by digital deconvolution and validated by immunofluorescence (n=30), illustrating that dysregulation of the ratio of Mf1 to Mf2 is an important factor contributing to implantation failure. Then, modules most associated with M1/M2 macrophages (Mfs) and their hub genes were then selected by weighted gene co-expression network and univariate analyses, then validated by GSE5099 macrophage dataset, qPCR analysis (n=16), and western blot. It revealed that closely related gene modules dominated three biological processes in macrophages: antigen presentation, interleukin−1−mediated signalling pathway, and phagosome acidification, respectively. Their hub genes were significantly altered in patients and related with ribosomal, lysosome, and proteasomal pathways. Finally, the artificial neural network (ANN) and nomogram models were established from hub genes, of which efficacy was compared and validated in the GSE165004 dataset (n=72). Models established by the selected hub genes exhibited excellent predictive values in both datasets, and ANN performed best with an accuracy of 98.3% and an AUC of 0.975 (95% CI 0.945-1). Conclusions: Macrophages, proven to be essential for endometrial receptivity, were regulated by gene modules dominating antigen presentation, interleukin−1−mediated signalling pathway, and phagosome acidification. Selected hub genes can effectively assess endometrial dysfunction receptivity for IVF outcomes by the ANN approach.

Pregnancy depends on a delicate balance of immune activation and regulation

It is well recognized that immune tolerance is important to prevent semiallografted fetuses from rejection by maternal immunocompetent cells; however, immune activation also plays an important role in placental development and fetal growth. Basic and clinical studies have shown that an imbalance between immune activation and regulation can lead to implantation failure, miscarriage, and preeclampsia. Here, the balance between immunostimulation and immunoregulation in reproduction will be reviewed.

Autophagy as a Therapeutic Target of Natural Products Enhancing Embryo Implantation

Infertility is an emerging health issue worldwide, and female infertility is intimately associated with embryo implantation failure. Embryo implantation is an essential process during the initiation of prenatal development. Recent studies have strongly suggested that autophagy in the endometrium is the most important factor for successful embryo implantation. In addition, several studies have reported the effects of various natural products on infertility improvement via the regulation of embryo implantation, embryo quality, and endometrial receptivity. However, it is unclear whether natural products can improve embryo implantation ability by regulating endometrial autophagy. Therefore, we performed a literature review of studies on endometrial autophagy, embryo implantation, natural products, and female infertility. Based on the information from these studies, this review suggests a new treatment strategy for female infertility by proposing natural products that have been proven to be safe and effective as endometrial autophagy regulators; additionally, we provide a comprehensive understanding of the relationship between the regulation of endometrial autophagy by natural products and female infertility, with an emphasis on embryo implantation.

Effect of Intrauterine Administration of Seminal Plasma for Patients with Recurrent Implantation Failure Before FET: Study Protocol for a Prospective Randomised Controlled Trial

Background: To assess the effect of intrauterine administration of seminal plasma for patients with recurrent implantation failure before frozen-thawed embryo transfer. Methods: Trial design: This is a parallel group, randomized (1:1 allocation ratio) controlled trial.Participants: All patients will be recruited from Chengdu Women’s and Children’s Central Hospital. Inclusion criteria: 1. Women after IVF/ICSI treatment in Chengdu Women’s and Children’s Central Hospital. 2.Infertile women with a history of recurrent implantation failure. 3.Infertile couples (both male and female) aged between 20 and 39 years;4. Couples who have at least 1 good quality embryos for transfer. 5. Males had negative in infectious disease test. 6. The males have semen. 7. Women who intend to undergo FET after IVF or ICSI or pre-implantation genetic testing for aneuploidy (PGT-A). 8. Competent and able to give informed consent. Intervention and comparator: Treatment group receiving intrauterine administration of seminal plasma before frozen-thawed embryo transfer. Main outcomes: Clinical pregnancy after frozen-thawed embryo transfer. Randomisation: Patients will be randomly allocated to either treatment or control group at 1:1 ratio. Random numbers will be generated by using software SPSS 25.0 performed by an independent statistician from Chengdu Women’s and Children’s Central Hospital. Blinding (masking): Only the data analyst will be blinded to group assignment. Numbers to be randomised (sample size): To account for a 10% dropout rate, we will recruit 174 patients (87 in each group). Trial status: The date of approval is 31rd May 2021, version 2.0. Recruitment started on 1rd June and is expected to end in July 2022. Discussion: Intrauterine administration of seminal plasma before frozen-thawed embryo transfer of patients with recurrent implantation failure may improve clinical pregnancy rate, it has great Page 2 of 14 significance for assisted reproduction. Trial registration: The study protocol has been approved by the ethics committees at Chengdu Women’s and Children’s Central Hospital. The trial was registered at the Chinese Clinical Trial Registry ChiCTR2100046803. Registered on 28 May 2021.

Prospective study to estimate the role of different infertility factors in prediction of unsuccessful IVF outcome

Research objective: in a prospective controlled study to investigate the role of HSP60, GroEl and other infertility factors as predictors of successful IVF outcome. Materials and methods. 106 female patients were divided into two groups: 54 individuals who received conventional treatment for infertility (using ICSI techniques for IVF) and 52 individuals who received conventional therapy with intravenous IgG, enoxaparin and aspirin. All collected blood samples were tested for HSP60 and GroEl antibodies using immunofluorescence and ELISA techniques at the time of admission, after treatment (and before embryo transfer), and after embryo transfer. We analyzed the factors that can be helpful as prognostic parameters to estimate the risk of implantation failure.Results. The risk of implantation failure is predicted when HSP60 level decreases from the first to the second measurement by less than 0.02 optical density units, with a sensitivity of 62% (95% confidence interval (CI) 47.2–75.3), and a specificity of 87.5% (95% CI 75.9–94.8), the positive predictive value was 81.6% (95% CI 68.2–90.2), the negative predictive value was 72.1% (95% CI 64.1–78.8). The GroEl value for the second dimension was more than 0.411 optical density units, which suggests a risk of treatment failure with a sensitivity of 64% (95% CI 49.2–77.1) and a specificity of 85.6% (95% CI 73.8–93.6), the positive predictive value was 80.0% (95% CI 67.1–88.7), the negative predictive value was 72.7% (95% CI 64.5–79.7). The highest (p < 0.05) value was observed at the beginning of treatment, and the lowest (p < 0.05) – during the third measurement.Treatment of the underlying cause of infertility led to a decrease in HSP60 and GroEl levels, which ensured a positive in vitro fertilization result. It was found that HSP60 and GroEl have a strong association with embryo implantation. The risk of implantation failure was strongly associated with twelve factors, the area under the curve (AUC) was 0.85 (95% CI 0.76–0.91).Conclusions. HSP60 and GroEl are good prognostic factors for predicting a successful IVF outcome in patients undergoing infertility treatment. The measurement of these parameters during the initial infertility examination may help in the immediate diagnosis of autoimmune infertility. Embryo implantation is a multifactorial process. The risk of implantation failure should be evaluated with multiple factors (twelve factors).

Recurrent Implantation Failure—Is It the Egg or the Chicken?

Recurrent implantation failure (RIF) is an undefined, quite often, clinical phenomenon that can result from the repeated failure of embryo transfers to obtain a viable pregnancy. Careful clinical evaluation prior to assisted reproduction can uncover various treatable causes, including endocrine dysfunction, fibroid(s), polyp(s), adhesions, uterine malformations. Despite the fact that it is often encountered and has a critical role in Assisted Reproductive Technique (ART) and human reproduction, RIF’s do not yet have an agreed-on definition, and its etiologic factors have not been entirely determined. ART is a complex treatment with a variable percentage of success among patients and care providers. ART depends on several factors that are not always known and probably not always the same. When confronted with repeated ART failure, medical care providers should try to determine whether the cause is an embryo or endometrium related. One of the most common causes of pregnancy failure is aneuploidy. Therefore, it is likely that this represents a common cause of RIF. Other RIF potential causes include immune and endometrial factors; however, with a very poorly defined role. Recent data indicate that the possible endometrial causes of RIF are very rare, thereby throwing into doubt all endometrial receptivity assays. All recent reports indicate that the true origin of RIF is probably due to the “egg”.

Insulin exposed endometrial epithelial cells cultured in a microfluidic device alters transcripts involved in translation that may contribute to reduced implantation capacity of the endometrium.

Obesity is a rapidly growing public health issue among women of reproductive age. It is also associated with decreased reproductive function including implantation failure. Implantation failure can result from a myriad of factors including impaired gametes and endometrial dysfunction. The mechanisms of how obesity-related hyperinsulinaemia disrupts endometrial function and implantation are poorly understood. Our study aims to investigate potential mechanisms by which insulin alters endometrial transcript expression, which may affect endometrial receptivity. Ishikawa cells mimicking human endometrial epithelium were seeded into a microfluidics organ-on-chip device to produce an in vitro endometrium. Syringe pump was attached to the microfluidics device to deliver three varying treatments into Ishikawa cells: 1) media control 2) vehicle control (PBS acidified to pH3 with acetic acid) 3) Insulin (2mg/mL) at a constant flow rate of 1uL/min for 24 hours to mimic secretion in vivo. Three biological replicates were obtained. Insulin-induced transcriptomic response of the in vitro endometrium was quantified via RNA sequencing, and subsequently analysed using DAVID and Webgestalt to identify Gene Ontology (GO) terms and signalling pathways. A Total of 29 transcripts showed differential expression levels across two comparison groups (control v vehicle control; vehicle control v insulin). There were nine transcripts significantly differentially expressed in vehicle control v insulin group (p<0.05). Functional annotation analysis of transcripts altered by insulin (n=9) identified three significantly enriched GO terms: SRP-dependent cotranslational protein targeting to membrane, poly(A) binding, and RNA binding (p<0.05). Over-representation analysis found three significantly enriched signalling pathways relating to insulin-induced transcriptomic response: protein export, glutathione metabolism, and ribosome pathways (p<0.05). Insulin-induced dysregulation of biological functions and pathways highlight potential mechanisms by which high insulin concentrations within maternal circulation may perturb endometrial receptivity.