記述言語：英語   出版者・発行元：Cancer Science  

BRCA1 is one of the causative genes for hereditary breast and ovarian cancer syndrome with a high risk of early-onset breast cancer. Whereas olaparib (OLA), an inhibitor of poly-ADP-ribose polymerase, has been applied as adjuvant therapy to those cancer patients, its effect on ovarian reproductive function remains unelucidated. Recently, a rat model (MUT; Brca1^(L63X/+) mutation) mimicking a human BRCA1 pathogenic variant has been established. Using this model, we evaluated the effects of OLA on ovarian reproductive function in comparison with the wild-type (WT) rats. MUT showed a significantly reduced number of primordial follicles and subfertility in accordance with aging. Oxidative stress was significantly elevated in the young MUT granulosa cells (GCs) accompanied by increased mTOR but decreased PTEN signals. OLA administration in MUT further decreased primordial follicles, with gene set enrichment analysis, indicating upregulated DNA repair pathways. Furthermore, a combination of OLA and cyclophosphamide (CPA) induced empty primordial follicles, recognized as CPA-induced severe ovarian toxicity. Whereas OLA + CPA caused greater reduction in primordial follicles both in MUT and WT in comparison with CPA alone, MUT ovaries were more susceptible to oxidative stress, potentially depleting primordial follicles via activation of GCs and inducing oocyte death due to accumulated DNA damage by OLA treatment. Our findings in this preclinical model underscore the importance of evaluating ovarian reserve prior to chemotherapy by performing reproductive consultation with female patients with BRCA1 pathogenic variants.

DOI： 10.1111/cas.16412

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PubMed

記述言語：英語   出版者・発行元：Reproductive Biology and Endocrinology  

Background: Endometriosis is an estrogen-dependent chronic inflammatory disease, however the mechanisms underlying inflammation remain unclear. Non-hormonal drugs that can prevent endometriosis progression and resolve endometriotic infertility are urgently required. We thus focused on cellular senescence as a novel feature of endometriosis. Senescent cells cause chronic inflammation via the senescence-associated secretory phenotype (SASP) factor. It has been reported the effects of senolysis for various diseases in recent years. The aim of this study was to validate the involvement of cellular senescence in endometriosis and as the effects of senolytic drug to develop a novel non-hormonal therapeutic strategy for endometriosis. Methods: The senescence markers were assessed by morphological features and semiquantitative immunofluorescence staining (senescence-associated b-galactosidase [SA-b-Gal], the cyclin-dependent kinase inhibitor 2 A locus [p16^INK4a], and laminB1) to compare among cell types (normal endometrial stromal cells [nESCs], endometrial stromal cells with endometriosis [eESCs], and ovarian endometriosis [OE] cyst-derived stromal cells [CSCs]). Expression of SASP markers was examined in cell culture supernatants using a cytokine array. In addition, the effects of senolytic drugs (azithromycin [AZM] and navitoclax [ABT263]) on endometriosis were evaluated in vitro and in vivo. The in vivo study used the endometriosis mice model. Results: CSCs exhibited stronger senescence markers. Semi-quantitative SA-β-Gal and p16^INK4a staining intensities were significantly increased, and that of LaminB1 was decreased in CSCs compared to those in nESCs and eESCs (SA-b-Gal, P < 0.001; p16^INK4a, P < 0.05; LaminB1, P < 0.05). Cytokine array analysis revealed elevated SASP-related cytokine levels, including interleukin-6 (IL-6), in CSC supernatants compared to those in nESCs. AZM and ABT263 reduced the viable fraction in CSCs (AZM: P < 0.001, ABT263: P < 0.01). Furthermore, AZM suppressed IL-6 expression in CSC culture supernatants (P < 0.05). In murine model, AZM administration reduced endometriotic lesion volume compared to that in vehicle (P < 0.05). Proliferative activity, IL-6 expression levels, and fibrosis within endometriotic lesions also decreased (Ki67, P < 0.01; IL-6, P < 0.001; fibrosis, P < 0.001). Conclusions: Our findings show that cellular senescence is involved in the pathogenesis of endometriosis and that AZM may be useful for preventing endometriosis progression by suppressing the secretion of IL-6 as a SASP.

DOI： 10.1186/s12958-025-01381-4

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記述言語：英語   出版者・発行元：Frontiers in Endocrinology  

Introduction: Endometriosis can cause of infertility, and evaluation methods for predicting clinical pregnancy outcomes are desired. Extracellular vesicles (EVs) exist in blood and it contains small non-coding RNAs (ncRNAs) that may reflect disease severity. In this study, we investigated small ncRNAs in serum EVs to identify specific biomarkers for predicting clinical pregnancy. Methods: Serum samples were collected from 48 patients who underwent assisted reproductive technology (ART). EVs were successfully isolated from serum samples and characterized using nanoparticle tracking assays, electron microscopy, and western blotting of EV’s markers. We performed small RNA sequencing and analyzed microRNA (miRNA) profiles in the infertility patients with and without endometriosis to detect pregnancy-predicting biomarkers. Results: Candidate miRNAs in serum EVs were selected by comparing patients without endometriosis who became pregnant (n = 13) with those who did not (n = 21). A total of 241 miRNAs were detected; however, no trends separated the two groups. Next, EVs from patients with endometriosis were analyzed and divided into pregnant (n = 4) and non-pregnant (n = 10) cases. Among the 224 candidate miRNAs, miRNA profiles of pregnant women with endometriosis were separated from those of non-pregnant women by receiver-operating characteristics (ROC) curve analysis (area under the curve [AUC] > 0.8). In patients with endometriosis, serum EVs may be useful for predicting possible pregnancy before infertility treatment. Finally, we used small RNA sequencing of the tissue to demonstrate that pregnancy-predicting miRNAs in serum EVs were produced from endometriosis lesions. Although no predictors were found from miRNAs in serum EVs without endometriosis, miRNAs in serum EVs of patients with endometriosis could provide novel noninvasive biomarkers to predict pregnancy and have potential clinical applicability in ART. Discussion: Further studies are required to examine the functional importance of these miRNAs to elucidate the pathological mechanisms of endometriosis and pregnancy.

DOI： 10.3389/fendo.2024.1442684

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記述言語：英語   出版者・発行元：Reproductive Medicine and Biology  

Purpose: Polycystic ovary syndrome (PCOS) significantly affects women. This study investigated the association between serum anti-Müllerian hormone (AMH) levels and menstrual cycle disorders, and AMH for PCOS in a general cohort of young Japanese women. Methods: We measured serum AMH levels in 528 healthy female students at two universities in Japan between 2014 and 2020. We investigated the association between serum AMH levels and hormone levels, menstrual cycle, and body mass index. Results: The mean (±standard deviation) AMH level was 4.78 ± 2.88 ng/mL. Correlations were observed between serum AMH and luteinizing hormone (LH) or LH/follicle-stimulating hormone (FSH) levels in women with irregular menstruation (LH: r = 0.542, p < 0.001; LH/FSH: r = 0.584, p < 0.001). The optimal serum AMH cutoff value that predicted LH ≥7.1 IU/L and LH/FSH ≥1.21 (PCOS diagnostic criteria revised by Japan Society of Obstetrics and Gynecology) in women with menstrual irregularities was 5.30 ng/mL (area under the curve: 0.815, sensitivity: 84.2%, specificity: 70.3%). Conclusions: Serum AMH can be measured during annual health checkups and may be a useful biomarker for early and arcuate diagnosis and intervention in women with PCOS.

DOI： 10.1002/rmb2.12615

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記述言語：英語   出版者・発行元：Reproductive Biology  

Ovarian endometrioma (OE) is a common gynecological disease that is often treated with surgery and hormonal treatment. However, ovarian cystectomy can impair the ovarian reserve (OR). Previously, we showed that perioperative administration of dienogest (DNG) is an effective option for OR preservation. However, there were differences in the extent of OR preservation among patients following perioperative DNG treatment. In the current study, we performed a global examination of serum microRNAs (miRNAs) to identify accurate biomarkers that predict post-operative restoration of OR following perioperative DNG treatment. We also sought to identify specific miRNAs related to the anti-Müllerian hormone (AMH). miRNA sequencing was performed on serum samples obtained from twenty-seven patients who received perioperative DNG treatment. Candidate miRNAs were selected by comparing patients whose ORs were restored postoperatively (responder group, n = 7) with those whose ORs were not (non-responder group, n = 7). miR-370–3p and miR-1307–3p were significantly upregulated in the responder group, whereas miR-27b-3p was upregulated in the non-responder group. The pretreatment value of each miRNA could predict DNG responsiveness for OR following ovarian cystectomy (area under the curve [AUC] > 0.8). The quantitative polymerase chain reaction (qPCR) revealed only miR-1307–3p was found to be significantly upregulated in the responder group (P < 0.05). In addition, we identified miR-139–3p, miR-140–3p, and miR-629–5p as AMH-associated miRNAs. The transition of AMH showed a correlation with miR-139–3p (P < 0.05, r = −0.76). The miRNAs identified herein represent potential serum biomarkers of clinical value in predicting OR prior to DNG treatment.

DOI： 10.1016/j.repbio.2023.100821

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記述言語：英語   出版者・発行元：Free Radical Research  

Pathogenic variants of BRCA1/2 constitute hereditary breast and ovarian cancer (HBOC) syndrome, and BRCA1/2 mutant is a risk for various cancers. Whereas the clinical guideline for HBOC patients has been organized for the therapy and prevention of cancer, there is no recommendation on the female reproductive discipline. Indeed, the role of BRCA1/2 pathogenic variants in ovarian reserve has not been established due to the deficiency of appropriate animal models. Here, we used a rat model of Brca2^{(p.T1942fs/+)} mutant of Sprague-Dawley strain with CRISPR-Cas9 editing to evaluate ovarian reserve in females. Fertility and ovarian follicles were evaluated and anti-Müllerian hormone (AMH) was measured at 8–32 weeks of age with a comparison between the wild-type and the mutant rats (MUT). MUT revealed a significantly smaller number of deliveries with fewer total pups. Furthermore, MUT showed a significant decrease in primordial follicles at 20 weeks and a low AMH level at 28 weeks. RNA-sequencing of the ovary at 10 weeks detected acceleration of the DNA damage repair pathway, which was accompanied by oxidative stress-induced DNA double-strand breaks, a decrease in PTEN, and an increase in mTOR in follicular granulosa cells. In conclusion, Brca2^{(p.T1942fs/+)} dissipates primordial follicles via early activation of granulosa cells through oxidative stress, leading to earlier termination of fertility.

DOI： 10.1080/10715762.2024.2320405

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記述言語：英語   出版者・発行元：Journal of Obstetrics and Gynaecology Research  

Aim: Both morbidity and mortality rates of cervical cancer are increasing, especially in reproductive-aged women. Radical trachelectomy (RT) is an effective fertility-preserving surgery for early-stage cervical cancer. This study aimed to determine the influence of RT on endometrial thickness during in vitro fertilization-embryo transfer (IVF-ET). Methods: Forty-four patients had undergone RT, and 23 women undergoing IVF-ET treatment (105 ET cycles) were included. Endometrial thickness during hormone replacement therapy (HRT) was retrospectively evaluated and compared between patients with and without RT. Results: Eleven patients (50 ET cycles) in the RT group and 12 (52 ET cycles) in the control group were investigated. Compared with the control group, higher ET cancellation rates were observed in patients in the RT group (1 of 52 cycles [control group] vs. 8 of 50 cycles [RT group], p < 0.01). Endometrial thinning was not affected by patient age at first IVF-ET treatment, history of artificial abortion, preservation of uterine arteries during RT, or postoperative chemotherapy (p = 0.27, 1, 1, and 1, respectively). Conclusions: Our data revealed that RT influenced endometrial thickness in IVF-ET. This was not affected by the background of the patients or perioperative management in this study. We could not reveal the underlying mechanism, but it is postulated that the transient postoperative uterine blood flow status and postoperative infections may have some effect on the endometrium. To resolve these issues, accumulation of evidences are required. We recommend informing patients about the impact of RT on IVF-ET before starting assisted reproductive technology (ART).

DOI： 10.1111/jog.15841

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記述言語：英語   出版者・発行元：Reproductive Sciences  

Polycystic ovary syndrome (PCOS), a common endocrine disorder, is associated with impaired oocyte development, leading to infertility. However, the pathogenesis of PCOS has not been completely elucidated. This study aimed to determine the differentially expressed genes (DEGs) and epigenetic changes in the oocytes from a PCOS mouse model to identify the etiological factors. RNA-sequencing analysis revealed that 90 DEGs were upregulated and 27 DEGs were downregulated in mice with PCOS compared with control mice. DNA methylation analysis revealed 30 hypomethylated and 10 hypermethylated regions in the PCOS group. However, the DNA methylation status did not correlate with differential gene expression. The pathway enrichment analysis revealed that five DEGs (Rps21, Rpl36, Rpl36a, Rpl37a, and Rpl22l1) were enriched in ribosome-related pathways in the oocytes of mice with PCOS, and the immunohistochemical analysis revealed significantly upregulated expression levels of Rps21 and Rpl36. These results suggest that differential gene expression in the oocytes of mice in PCOS is related to impaired folliculogenesis. These findings improve our understanding of PCOS pathogenesis.

DOI： 10.1007/s43032-022-01095-7

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記述言語：英語   出版者・発行元：Reproductive Medicine and Biology  

Background: Primary ovarian insufficiency (POI) is characterized by the development of hypergonadotropic hypogonadism before 40 years of age and leads to intractable infertility. Although in vitro fertilization and embryo transfer with donated eggs enables pregnancy, not a few patients desire pregnancy using their oocytes. However, follicular development is rare and unpredictable in patients with POI. Thus, there is a need for treatments that promote the development of residual follicles and methods to accurately predict infrequent ovulation. Methods: This review discusses the effects of various treatments for obtaining eggs from POI patients. Furthermore, this study focused a potential marker for predicting follicular growth in patients with POI. Main Findings: Different treatments such as hormone-replacement therapy, dehydroepiandrosterone supplementation, platelet-rich plasma injection, and in vitro activation have shown varying degrees of effectiveness in retrieving oocytes from patients with POI. To predict follicle development in the cycle, elevated serum estradiol and reduced follicle-stimulating hormone (FSH) levels are important. However, these markers are not always reliable under continuous estradiol-replacement therapy. As a novel marker for predicting follicle growth, serum anti-Müllerian hormone (AMH) levels, measured using the picoAMH enzyme-linked immunosorbent assay, were found to predict follicle growth in patients and the cycle. Conclusion: This review highlights the challenges and available interventions for achieving pregnancy using a patient's oocytes in cases of POI. We believe that a combination of currently available treatments and prediction methods is the best strategy to enable patients with POI to conceive using their own eggs. Although AMH levels may predict follicle growth, further research is necessary to improve the chances of successful follicular development and conception in patients with POI.

DOI： 10.1002/rmb2.12556

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Web of Science

担当区分：筆頭著者  

DOI： 10.1038/s41598-022-15564-1

Open Access

DOI： 10.1186/s12958-022-00924-3

Open Access

記述言語：英語  

DOI： 10.1186/s12958-021-00866-2

Open Access

開催年月日： 2024年2月

会議種別：口頭発表（一般）