Single-cell transcriptome analysis reveals endometrial immune microenvironment in minimal/mild endometriosis.

Endometriosis is a common inflammatory disorder in women of reproductive age due to an abnormal endometrial immune environment and is associated with infertility. This study aimed to systematically understand the endometrial leukocyte types, inflammatory environment, and impaired receptivity at single-cell resolution. We profiled single-cell RNA transcriptomes of 138 057 endometrial cells from endometriosis patients (n = 6) and control (n = 7), respectively, using 10x Genomics platform. We found that one cluster of epithelial cells that expressed PAEP and CXCL14 was mostly from the control during the window of implantation (WOI). This epithelial cell type is absent in the eutopic endometrium during the secretory phase. The proportion of endometrial immune cells decreased in the secretory phase in the control group, whereas the cycle variation of total immune cells, NK cells, and T cells was absent in endometriosis. Endometrial immune cells secreted more IL-10 in the secretory phase than in the proliferative phase in the control group; the opposite trend was observed in endometriosis. Proinflammatory cytokines levels in the endometrial immune cells were higher in endometriosis than in the control group. Trajectory analysis revealed that the secretory phase epithelial cells decreased in endometriosis. Ligand-receptor analysis revealed that 11 ligand-receptor pairs were upregulated between endometrial immune and epithelial cells during WOI. These results provide new insights into the endometrial immune microenvironment and impaired endometrial receptivity in infertile women with minimal/mild endometriosis.

Gestational metformin administration in women with polycystic ovary syndrome: A systematic review and meta-analysis of randomized control studies.

AIMS: To evaluate metformin's effects on pregnancy outcomes in women with polycystic ovary syndrome. METHODS: A literature search was conducted using PubMed, EMBASE, Web of Science, MEDLINE, and the Cochrane Library. All randomized controlled trials comparing metformin administration during pregnancy versus placebo or blank in PCOS women were selected. The primary outcomes were the incidence of gestational diabetes mellitus (GDM), preterm delivery, and miscarriage. We combined data with the Review Manager. Bayesian meta-analysis was employed for further verification with the R software. RESULTS: Six randomized control trial studies involving 1229 participants were included. Metformin use was associated with reduced risk of preterm delivery (Risk ratios [RR], 0.45; 95% CI, 0.25-0.80; p, 0.007) and higher larger neonatal head circumference (Mean difference (MD), 0.47; 95% CI, 0.20-0.74; p, 0.0006] but had no effect on the incidence of GDM (RR 1.87; 95% CI, 0.58-1.87; p, 0.89), miscarriage (RR, 0.85; 95% CI, 0.45-1.60; p, 0.62), pre-eclampsia (RR, 1.18; 95% CI, 0.43-3.21; p, 0.75), neonatal length (MD, 0.33; 95% CI, -0.12-0.78; p, 0.15) and birthweight (MD, 73.78; 95% CI, -52.98-200.53; p, 0.17). CONCLUSIONS: Metformin administration in PCOS pregnancies was associated with reduced preterm delivery risk and larger neonatal head circumference.

Expression of programmed death-1 (PD-1) and its ligand PD-L1 is upregulated in endometriosis and promoted by 17beta-estradiol.

The programmed death-1 (PD-1)/PD-L1 pathway plays important roles in immune responses and peripheral tolerance. Under pathological conditions, this pathway can suppress protective T cell responses and induce immune system disorders. Endometriosis (EM) is an estrogen-dependent, immune-associated disease. The expression of PD-1 and PD-L1 has been studied in a variety of cancers and autoimmune diseases. Few studies, however, have attempted to explore their expression and the possibility that they are regulated by estrogen in EM. Eutopic and ectopic endometria and blood samples were collected from 15 women with EM and 15 women without EM. PD-1/PD-L1 expression in endometrial tissues was detected using immunohistochemistry and western blot analysis. Their expression in blood was detected using flow cytometry. In addition, their expression was evaluated in endometrial cells after estrogen and cytokine treatment. We observed PD-1/PD-L1 expression in both eutopic and ectopic endometria, with elevated expression in EM. Their expression was also higher in CD4+/CD8+ T cells in blood from EM patients. In addition, treatment with 17ß-estradiol upregulated PD-L1 expression in eutopic epithelial cells in EM. These data suggest that the PD-1/PD-L1 pathway may be involved in EM immune dysfunction and be regulated by estrogen.

Clinical utility of office hysteroscopy following failed in vitro fertilization-embryo transfer: A retrospective cohort study.

OBJECTIVE: Despite its widespread use, in vitro fertilization (IVF) outcomes are challenged by implantation failure, largely due to factors such as embryo quality and endometrial receptivity. In this study, we investigated the clinical effect of office hysteroscopy (OH) on the subsequent frozen-thawed embryo transfer (FET) in infertile women who experienced a failed IVF-embryo transfer (IVF-ET) cycle. METHODS: We included 577 infertile women who underwent OH because of a history of failed ET between October 2019 and September 2021. During OH, visible endometrial polyps (EPs) were diagnosed and removed by curette or biopsy forceps; chronic endometritis (CE) was diagnosed by histopathology and immunohistochemistry and treated with oral doxycycline (0.2 g/d) for 14 days. According to the hysteroscopic findings and endometrial pathology with immunohistochemistry, patients were divided into three groups: group A (n = 161) had CE with or without EPs, group B (n = 156) had EPs only, and group C (n = 260) had no CE or EPs. RESULTS: In the following FET cycle, the implantation rates were 47%, 51%, and 45% (P = 0.411); the clinical pregnancy rates were 56%, 62%, and 55% (P = 0.436); the live birth rates were 45%, 51%, and 42% (P = 0.205); and the miscarriage rates were 18%, 16%, and 22% (P = 0.497) in groups A, B, and C, respectively. There were no significant differences among groups (P > 0.05). CONCLUSION: OH is helpful for diagnosis and treatment of abnormal intrauterine environment in women with a failed IVF cycle and further improves their pregnancy outcome in the following FET.

Existence of chronic endometritis and its influence on pregnancy outcomes in infertile women with minimal/mild endometriosis.

OBJECTIVE: This study aimed to evaluate the prevalence of chronic endometritis (CE) in women with minimal/mild endometriosis and to analyze whether CE affects their pregnancy outcomes. METHODS: This retrospective study included 201 infertile women who were diagnosed with minimal/mild endometriosis after undergoing hysteroscopy combined with laparoscopy from January 2016 to December 2018. Immunohistochemistry was used to detect CD138 and CD38, which are specific markers of plasma cells in the endometrial stroma to diagnose CE. Subsequently, we investigated the prevalence of CE and the effects of CE on spontaneous cumulative pregnancy rate, live birth rate, and miscarriage rate within 24 months after surgery. RESULTS: The prevalence of CE in infertile women with minimal/mild endometriosis was 24.38%. Patients diagnosed with CE showed a significantly lower cumulative pregnancy rate and live birth rate compared with women without CE (46.51% vs. 71.13% [P = 0.004]; 44.19% vs. 63.38% [P = 0.025]). However, the rate of miscarriage in women with CE was also lower than in women without CE (0 vs. 7.04%, P = 0.074). CONCLUSION: Since CE had an adverse effect on cumulative pregnancy rate and live birth rate in infertile women with minimal/mild endometriosis, we suggested that diagnosis and treatment of CE may improve their pregnancy outcomes.

Increased Expression of TGF-ß1 Contributes to the Downregulation of Progesterone Receptor Expression in the Eutopic Endometrium of Infertile Women with Minimal/Mild Endometriosis.

Endometriosis is a hormone-dependent disease associated with impaired immunoregulation. In our recent study, we have characterized the trascriptomic transformation of eutopic endometrium from patients with minimal/mild endometriosis and controls across the menstrual cycle. However, the regulatory mechanism of altered immune microenvironment in eutopic endometrial stromal cells (ESCs) remains unclear. Here, we want to explore the regulation of immune cell to progesterone resistance and endometrial receptivity in the eutopic ESCs by cytokine (TGF-ß1), and to understand the effect of TGF-ß1 on the decidualization of the eutopic ESCs. Primary culture of eutopic ESCs was performed to explore the effects of TGF-ß1 on the expression of Smad and progesterone receptor (PR) and the in vitro decidualization. Additionally, co-immunoprecipitation (Co-IP) was used to explore the direct interaction between Smad and PR. We found an attenuate expression of PRB protein (p=0.026) after using TGF-ß1 in eutopic ESCs, although the difference of PRA before and after treatment was not significant (p=0.678). Similarly, the results of qRT-PCR showed that the mRNA level of PR (p<0.001), PRB (p=0.003) and HOXA10 (p<0.001) decreased significantly after TGF-ß1 treatment, but that increased (p<0.023, for all) after SB431542 treatment in the eutopic ESCs. Moreover, TGF-ß1 has a negative effect on the in vitro decidualization of eutopic ESCs (p=0.003). And the group with treatment of both TGF-ß1 and SB435142 in eutopic ESCs showed significant decidual-like changes with increased prolactin level (p=0.01). We did not observe any physical interaction between the PR and p-Smad3/Smad3 proteins by using Co-IP. By activating TGF-ß/Smad signaling in eutopic ESCs, elevated TGF-ß1 from CD45+ immune cells could attenuate expression of PR, and further decrease endometrial receptivity.

The Microbial Composition of Lower Genital Tract May Affect the Outcome of in vitro Fertilization-Embryo Transfer.

Objective: This work was conducted in order to study the effect of the lower genital tract (vaginal and cervical canal) microbiota on pregnancy outcomes of reproductive-aged women receiving embryo transfer. Study design: A total of 150 reproductive-aged patients who received the first fresh in vitro fertilization-embryo transfer (IVF-ET) were included in the study. Samples from the vagina and cervical site of each patient were collected separately using sterile swabs before ET. Genomic DNA was pyrosequenced for the V3-V4 regions of the 16S ribosomal RNA gene. Further bioinformatics analysis was performed using QIIME and R package. Pregnancy outcomes were followed and analyzed to compare differences in microbial composition. Results: The cervical microbiota had a higher Shannon index than the vaginal microbiota, and the microbial composition was different between the two sites. However, the Sorenson index between the two sites within the same individual was 0.370 (0.309-0.400). A total of 89 patients achieved clinical pregnancy after ET, while 61 failed. The Shannon indices and the microbial community of both vaginal and cervical microbiota between pregnant and non-pregnant groups were not significantly different. The relative abundance of Lactobacillus in the vagina and cervical canal did not differ between the two groups. Linear discriminant analysis, random forest analysis, and receiver-operating characteristic curve analysis showed that Bifidobacterium, Prevotella, and Lactobacillus iners in the vagina, as well as Solanum torvum, Fusobacterium, and Streptococcus in the cervix, may be negatively associated with clinical pregnancy after IVF. Conclusion: The cervical microbiota was more diverse than the vaginal microbiota, but because of anatomical continuity, there was a correlation between the two sites. The microbial composition of the vagina and cervical canal may influence the outcome of IVF-ET, but more samples are needed to verify this conclusion.