Gene expression signatures associated with chronic endometritis revealed by RNA sequencing

IntroductionChronic endometritis (CE) is a persistent inflammatory condition of the endometrium characterized by the infiltration of plasma cells in the endometrial stroma. CD138 immunohistochemistry is considered to improve the CE diagnosis rate.MethodsUsing the number of CD138-positive cells equal or greater than five as a diagnostic criterion for CE, we identified 24 CE and 33 non-CE cases among women with infertility. We conducted RNA-sequencing analysis for these 57 cases in total as an attempt to elucidate the molecular pathogenesis of CE and to search for new biomarkers for CE.Results and DiscussionBy comparing CE and non-CE groups, we identified 20 genes upregulated in the endometria of CE patients, including 12 immunoglobulin-related genes and eight non-immunoglobulin genes as differentially expressed genes. The eight genes were MUC5AC, LTF, CAPN9, MESP1, ACSM1, TVP23A, ALOX15, and MZB1. By analyzing samples in the proliferative and secretory phases of the menstrual cycle separately, we also identified four additional non-immunoglobulin genes upregulated in CE endometria: CCDC13 by comparing the samples in the proliferative phase, and OVGP1, MTUS2, and CLIC6 by comparing the samples in the secretory phase. Although the genes upregulated in CE may serve as novel diagnostic markers of CE, many of them were upregulated only in a limited number of CE cases showing an extremely high number of CD138-positive cells near or over one hundred. Exceptionally, TVP23A was upregulated in the majority of CE cases regardless of the number of CD138-positive cells. The upregulation of TVP23A in the endometria of CE cases may reflect the pathophysiology of a cell-type or cell-types intrinsic to the endometrium rather than the accumulation of plasma cells. Our data, consisting of clinical and transcriptomic information for CE and non-CE cases, helped us identify gene expression signatures associated with CE

Clinical strategies for ART treatment of infertile women with advanced maternal age

Abstract Background An ever‐increasing number of women in our country with advanced maternal age are choosing to achieve pregnancy. This means effective strategies are needed for infertile patients. Questions arise, however, concerning the need for ovarian stimulation, and, if so, whether intracytoplasmic sperm injection (ICSI) is better than conventional insemination for those women who may have only one mature oocyte. Methods We evaluated our data to answer these questions. Herein, we also introduce our strategy for patients who show unsynchronized follicular growth. Main findings Ovarian stimulation in ART treatment for patients with advanced maternal age has resulted in the achievement of higher pregnancy rates, and therefore, this form of stimulation is often selected. Based on our data, ICSI as an insemination procedure has not improved clinical pregnancy rates compared with conventional insemination and has actually decreased the clinical pregnancy rates. Conclusion In this article, we reviewed and compared the protocols and strategies that are available to increase the number of developed embryos for the patients with advanced maternal age. We hope that this review will be helpful for both patients and clinicians

#44 : Effects of Optimum (Optimization of Thyroid, Thrombophilia, Immunity, and Uterine Milieu) Treatment Strategy on Euploid Blastocyst Transfer in Advanced Aged Women with Recurrent Reproductive Failure

Background and Aims: Does the Optimization of Thyroid function, Thrombophilia, Immunity and Uterine Milieu (OPTIMUM) treatment strategy contribute to improving pregnancy outcomes after single euploid blastocyst transfer (SEBT) in patients with a history of repeated implantation failure (RIF) and/or recurrent pregnancy loss (RPL)? Method: Between January 2019 and May 2022, women aged ≥ 40 years with RIF after three or more embryo transfer using morphology good embryos and/or RPL after two or more clinical pregnancy losses underwent preimplantation genetic testing for aneuploidy (PGT-A) and RIF/RPL testing, including a hysteroscopy, endometrial biopsy for CD138 immunostaining and bacterial culture, and serum 25-hydroxyvitamin D3, interferon-γ-producing helper-T (Th1) cell, IL-4-producing helper-T (Th2) cell, thyroid-stimulating hormone, thyroid peroxidase antibody, and thrombophilia screening. We treated chronic endometritis with antibiotics, high Th1/Th2 cell ratios with vitamin D and/or tacrolimus, overt/subclinical hypothyroidism with levothyroxine, and thrombophilia with low-dose aspirin. Of 160 consecutive women who underwent SEBT, we compared 127 and 33 women with and without the OPTIMUM treatment strategy, respectively. Results: RIF/RPL testing identified intrauterine abnormalities in 67 (52.8%), aberrant high Th1/Th2 cell ratios in 38 (29.9%), thyroid dysfunction in 19 (15.0%), and thrombophilia in 24 (18.9%). The clinical pregnancy and live birth rates in the OPTIMUM group was significantly higher than that in the control group (73.9% and 45.5%, respectively; p = 0.005 and 64.7% and 39.4%, respectively; p = 0.01) in women with RIF (Table 1). Whereas there was no significant difference of miscarriage rate in RPL women with and without OPTIMUM (5.5% and 13.3%, respectively; p = 0.58). Conclusion: In the women aged ≥40 years with RIF who underwent PGT-A, the OPTIMUM treatment strategy improved pregnancy outcomes after SEBT. In RPL, however, both PGT-A with and without OPTIMUM resulted in low miscarriage rates and no significant difference was recognized

#95 : Analysis of the Incidence of and Risk Factors for Chronic Endometritis Recurrence in Infertile Women

Background and Aims: Chronic endometritis (CE) is an inflammatory condition of the endometrium which is related to repeated implantation failure. Recovery from CE can increase opportunities for successful pregnancy; however, some of them remain incapable of conceiving or have a miscarriage and concern CE recurrence. The aim of this study is to identify the incidence rates of and risk factors for the recurrence of CE in infertile women. Method: The study population consisted of 1,897 infertile women from a hospital specializing in reproductive medicine who recovered from CE between December 2018 and August 2021. Among the 152 women (13.0%) who did not conceive or experienced pregnancy loss, 105 consecutive women who underwent repeat endometrial biopsy for CD138 immunostaining and endometrial bacterial culturing within 18 months from CE recovery were recruited. Thereafter, patients with and without CE recurrence were compared. Results: The total recurrence rate of CE was 29.5% (31 women). Multivariable logistic regression analysis to determine risk factors for CE recurrence revealed that hysteroscopic surgery (odds ratio [OR], 0.10; 95% confidence interval [CI], 0.02–0.56) and pregnancy loss (OR, 4.13; 95% CI, 1.31–13.05) were significantly associated with decreased and increased CE recurrence rates, respectively. Also, re-examination with CD138 immunostaining after 16–18 months (OR, 9.75; 95% CI, 1.47–64.64) was significantly associated with increased CE recurrence rates. Among 40 patients without a history of hysteroscopic surgery and pregnancy loss, the cumulative CE recurrence rates after 6, 12, and 18 months were 12.5%, 23.3%, and 30.0%, respectively (Graph 1). Conclusion: We recommend re-examination with endometrial CD138 immunostaining in patients with pregnancy loss or long-term infertility during fertility treatment. Hysteroscopic surgery without antibiotic therapy for CE associated with intrauterine abnormalities is also recommended

Clinical outcomes of assisted reproductive technology treatment by using a self‐injection of recombinant human chorionic gonadotropin as the final maturation trigger

Abstract Purpose To evaluate the efficacy and safety of self‐injections of the prefilled recombinant human chorionic gonadotropin (r‐hCG) in a syringe in assisted reproductive technology (ART) treatment for the maturation trigger (MT), as compared to self‐injections of conventional hCG and intranasal administration of gonadotropin‐releasing hormone agonist (GnRH‐a). Methods Between January and April, 2017, 396 patients who underwent oocyte retrieval were recruited. Of these, 396 patients were classified into three groups, according to the types of MT: (1) the urinary human chorionic gonadotropin (u‐hCG) group that consisted of patients who had a self‐injection of u‐hCG (n = 127); (2) the GnRH‐a group that received nasal administration of GnRH‐a (n = 159); and (3) the r‐hCG group that had a self‐injection of r‐hCG (n = 110). Several ART outcomes were evaluated. Results The mature oocyte retrieval rate was not different between the u‐hCG, r‐hCG, and GnRH‐a groups and the fertilization and cleavage rates were similar between the three groups. The clinical pregnancy rates did not significantly differ between the GnRH‐a group and the u‐hCG group; however, it was significantly lower in the GnRH‐a group, compared to the r‐hCG group. No difference was observed in the incidence of moderate or more severe ovarian hyperstimulation syndrome among the three groups. Conclusion The self‐injection of the prefilled r‐hCG is a favorable MT for ART patients

Parental age and gene expression profiles in individual human blastocysts

Abstract The epigenetic status of the genome changes dynamically from fertilization to implantation. In addition, the physiological environment during the process of gametogenesis, including parental age, may affect the epigenome of the embryo after fertilization. It is important to clarify the influence of parental age on gene expression in the embryo in terms of transgenerational epigenetics to improve the techniques currently used in assisted reproductive medicine. Here, we performed single-embryo RNA-seq analysis on human blastocysts fertilized by intracytoplasmic sperm injection, including from relatively elderly mothers, to elucidate the effects of parental age on the embryonic gene expression profile. We identified a number of genes in which the expression levels were decreased with increasing maternal age. Among these genes, several are considered to be important for meiotic chromosomal segregation, such as PTTG1, AURKC, SMC1B and MEIKIN. Furthermore, the expression levels of certain genes critical for autophagy and embryonic growth, specifically GABARAPL1 and GABARAPL3, were negatively correlated with advanced paternal age. In addition, levels of transcripts derived from major satellite repeats also decreased as the maternal age increased. These results suggest that epigenetic modifications of the oocyte genome may change with parental age and be transmitted to the next generation

Vitamin D Regulates Maternal T-Helper Cytokine Production in Infertile Women

Vitamin D (VD) deficiency is associated with reproductive failure. However, the relationship between VD and maternal immunity remains unclear. We investigated the clinical efficacy of VD in maternal T-helper (Th) cytokines in 276 infertile women and examined for Th1 and Th2 cells based on the deficient, insufficient, and sufficient serum 25-hydroxyvitamin D3 (25[OH]VD) levels (<12, 12–30, and >30 ng/mL, respectively). Most infertile women had a low-level of VD (87.3%). Immunological tests of pre-/post-VD supplementation were performed in patients who were deficient and insufficient in VD. Of 23 patients, 11 (47.8%) exhibited sufficient VD levels after supplementation. Th1/Th2 cell ratio in patients with insufficient VD was significantly decreased after supplementation (p = 0.004). After supplementation, serum 25(OH)VD levels of the patients: 11 in the sufficient group showed significant decreases in Th1 cell level and Th1/Th2 cell ratio (p = 0.032 and 0.010, respectively), whereas no significant differences in Th1/Th2 cell ratio were recognized in the insufficient group. Furthermore, mid-luteal endometrial biopsies (n = 18) were processed for primary cultures and measured interferon [IFN]-γ and interleukin [IL]-4 in condition media. Decidualizing cultures with 1,25-dihydroxvitamin D3 (1,25[OH]2VD) decreased IFN-γ. Sufficient VD supplementation in women with insufficient VD may optimize maternal T-helper cytokines during pregnancy via rebalancing the Th1/Th2 cell ratio

Therapeutic effects of an oral gonadotropin‐releasing hormone receptor antagonist, relugolix, on preventing premature ovulation in mild ovarian stimulation for IVF

Abstract Purpose Can relugolix, a novel oral gonadotropin‐releasing hormone receptor (GnRH) antagonist, function as an alternative ovulation inhibitor to GnRH antagonist injections? Methods This single‐center, cross‐sectional retrospective study compared premature ovulation rates and clinical outcomes in IVF treatment after mild ovarian stimulation with 40 mg of relugolix (relugolix group) or 0.25‐mg injections of ganirelix acetate or cetrorelix acetate (injection group) between March 2019 and January 2020. Of 247 infertile women (256 IVF cycles) aged ≤42 years, 223 women (230 cycles) were evaluated. In the relugolix and injection groups, we compared 104 and 85 cycles after GnRH antagonist use before the LH surge (LH levels <10 mIU/ml) and 22 and 19 cycles during the LH surge (LH levels ≥10 mIU/ml), respectively. Results Before the LH surge, the ovulation rates in the two groups were very low (p = 0.838), however; during the LH surge, the cycles using relugolix had a high ovulation rate of 40.9% compared with no ovulation in the injection group (p = 0.002). There were no significant differences in embryo culture findings and pregnancy outcomes between the two groups. Conclusions Although relugolix had a high ovulation suppressive effect, when the LH surge occurred, its effect was insufficient