研究者詳細 - 中村　智子

Novel ovarian endometriosis model causes infertility via iron-mediated oxidative stress in mice Open Access

Hayashi S., Nakamura T., Motooka Y., Ito F., Jiang L., Akatsuka S., Iwase A., Kajiyama H., Kikkawa F., Toyokuni S.

Redox Biology 37 巻頁： 101726 2020年10月

出版者・発行元：Redox Biology

Ovarian endometriosis (OE) provides women of reproductive age with not only severe menstrual pain but also infertility and an increased risk for ovarian carcinogenesis. Whereas peritoneal endometriosis models have been developed with syngeneic implantation of minced uterine tissue and oncogenic K-ras allele with conditional Pten deletion within ovarian surface epithelium generated preneoplastic endometrial glandular morphology, followed by endometrioid adenocarcinoma, there has been no mouse model of OE similar to human counterparts, applicable to preclinical studies. Here we for the first time established a murine OE model that reveals infertility, and evaluated the involvement of iron catalyzed oxidative stress in the pathogenesis. Minced uterine tissue from female mice was implanted on ovarian surface of syngeneic mice after bursectomy to induce OE. Ectopic growth of endometrium was observed in association with ovary 4 weeks after implantation in 85.7% (12/14) of the operated mice with our protocol. Endometriotic lesions involved intestine, pancreas and peritoneal wall. Fibrosis around the ovary was prominent and increased time-dependently in the OE group. Iron accumulation was significantly increased in the OE group, leading to oxidative stress in each stage of the follicles as evaluated by 4-hydroxy-2-nonenal-modified proteins and 8-hydroxy-2′-deoxyguanosine. Expression of follicle stimulating hormone receptor in the follicles revealed a significant decrease during pre-antral, antral and pre-ovulatory phases in the OE group. Finally, the number of pups was significantly reduced in the OE group in comparison to the controls. This model affords an opportunity to evaluate agents or procedures to counteract ovarian endometriosis in the preclinical settings.

DOI： 10.1016/j.redox.2020.101726

Mutant FOXL2C134W hijacks SMAD4 and SMAD2/3 to drive adult granulosa cell tumors Open Access

Weis-Banke S.E., Lerdrup M., Kleine-Kohlbrecher D., Mohammad F., Sidoli S., Jensen O.N., Yanase T., Nakamura T., Iwase A., Stylianou A., Abu-Rustum N.R., Aghajanian C., Soslow R., da Cruz Paula A., Koche R.P., Weigelt B., Christensen J., Helin K., Cloos P.A.C.

Cancer Research 80 巻 ( 17 ) 頁： 3466 - 3479 2020年9月

出版者・発行元：Cancer Research

The mutant protein FOXL2C134W is expressed in at least 95% of adult-type ovarian granulosa cell tumors (AGCT) and is considered to be a driver of oncogenesis in this disease. However, the molecular mechanism by which FOXL2C134W contributes to tumorigenesis is not known. Here, we show that mutant FOXL2C134W acquires the ability to bind SMAD4, forming a FOXL2C134W/SMAD4/SMAD2/3 complex that binds a novel hybrid DNA motif AGHCAHAA, unique to the FOXL2C134W mutant. This binding induced an enhancer-like chromatin state, leading to transcription of nearby genes, many of which are characteristic of epithelial-to-mesenchymal transition. FOXL2C134W also bound hybrid loci in primary AGCT. Ablation of SMAD4 or SMAD2/3 resulted in strong reduction of FOXL2C134W binding at hybrid sites and decreased expression of associated genes. Accordingly, inhibition of TGFb mitigated the transcriptional effect of FOXL2C134W. Our results provide mechanistic insight into AGCT pathogenesis, identifying FOXL2C134W and its interaction with SMAD4 as potential therapeutic targets to this condition.

DOI： 10.1158/0008-5472.CAN-20-0259

Focal Adhesion Kinase-Mediated Sequences, Including Cell Adhesion, Inflammatory Response, and Fibrosis, as a Therapeutic Target in Endometriosis

Nagai T., Ishida C., Nakamura T., Iwase A., Mori M., Murase T., Bayasula , Osuka S., Takikawa S., Goto M., Kotani T., Kikkawa F.

Reproductive Sciences 27 巻 ( 7 ) 頁： 1400 - 1410 2020年7月

出版者・発行元：Reproductive Sciences

Endometriosis has several distinguishing features in the ectopic endometrium, including chronic inflammation and fibrosis. According to the retrograde menstruation theory, endometriotic cells are derived from eutopic endometrial cells, and adhesion of endometrial cells to the extracellular matrix can be the initial step in the development of endometriosis. Therefore, we hypothesized that cell adhesion, which mediates a sequence of events in the development of endometriosis triggering inflammatory responses and tissue fibrosis could be a possible therapeutic target for endometriosis. We found co-upregulation of focal adhesion kinase (FAK) and monocyte chemoattractant protein-1 (MCP-1) in the endometriotic tissues compared with that in the normal endometrium. MCP-1 secretion was significantly higher in the endometriotic stromal cells than in the eutopic endometrial stromal cells. Furthermore, co-culture of U937 cells and endometriotic stromal cells upregulated secretion of transforming growth factor-β1 (TGF-β1). A FAK inhibitor significantly inhibited the secretion of MCP-1 in the endometriotic stromal cells and TGF-β1 in the co-culture with macrophages. FAK inhibitor treatment in the murine endometriosis model demonstrated a decrease in the formation of endometriotic lesions as well as the expression of MCP-1 and TGF-β1. Our results suggest that the FAK-mediated sequential development of endometriosis, including inflammatory response and tissue fibrosis, can be a new therapeutic target in endometriosis.

DOI： 10.1007/s43032-019-00044-1

Primary Follicles Suppress the Growth of Primordial Follicles.

Wei Wei, Komatsu Kouji, Murase Tomohiko, Sonehara Reina, Miyake Natsuki, Murakami Mayuko, Ganieva Umida, Bayasula Bayasula, Yoshita Sayako, Muraoka Ayako, Hayashi Shotaro, Nakanishi Natsuki, Kasahara Yukiyo, Takasaki Nobuyoshi, Nakamura Tomoko, Osuka Satoko, Goto Maki, Kikkawa Fumitaka

REPRODUCTIVE SCIENCES 27 巻 ( SUPPL 1 ) 頁： 283A - 283A 2020年3月

The Function of POTEF in Human Granulosa Cells, an Antigen for Ovarian Autoimmunity, in POI Patients.

Kasahara Yukiyo, Osuka Satoko, Takasaki Nobuyoshi, Wei Wei, Nakanishi Natsuki, Bayasula Bayasula, Nakamura Tomoko, Murase Tomohiko, Goto Maki, Kikkawa Fumitaka

REPRODUCTIVE SCIENCES 27 巻 ( SUPPL 1 ) 頁： 199A - 199A 2020年3月

Involvement of Transcription Factor 21 in the Pathogenesis of Fibrosis in Endometriosis Open Access

Ganieva U., Nakamura T., Osuka S., Bayasula , Nakanishi N., Kasahara Y., Takasaki N., Muraoka A., Hayashi S., Nagai T., Murase T., Goto M., Iwase A., Kikkawa F.

American Journal of Pathology 190 巻 ( 1 ) 頁： 145 - 157 2020年1月

出版者・発行元：American Journal of Pathology

Repeated tissue injury and repair and fibrosis play a pivotal role in endometriosis. Fibrotic tissue consists of extracellular matrix proteins, regulated by transcriptional factors promoting cell proliferation and survival. Periostin is one of the putative key extracellular matrix proteins. This study aimed to determine whether transcription factor 21 (TCF21) is involved in the development of endometriosis as an upstream regulatory gene of periostin. Formalin-fixed, paraffin-embedded tissue samples [normal endometrium of women without endometriosis; eutopic endometrium of women with endometriosis; ovarian endometriosis (OE); and deep infiltrating endometriosis (DIE)] and respective cells were analyzed. Basal, transiently stimulated, and knocked down periostin and TCF21 concentrations in stromal cells of women with or without endometriosis were examined. Periostin and TCF21 expressions were undetected in normal endometrium of women without endometriosis, weakly positive in eutopic endometrium of women with endometriosis, moderately positive in OE, and strongly positive in DIE. Type 2 helper T-cell cytokines (IL-4, IL-13, and transforming growth factor-β1) increased the mRNA expression of periostin and TCF21. These cytokines, periostin, and TCF21 colocalized in the stroma of OE and DIE. siRNA against human TCF21 gene suppressed periostin expression. Transfection of TCF21 plasmid vector into stromal cells of women without endometriosis, which originally expressed neither periostin nor TCF21, resulted in TCF21 and periostin expression. TCF21 and periostin are involved in the regulation of fibrosis in endometriosis. TCF21 may be a promising therapeutic target and biomarker in endometriosis.

DOI： 10.1016/j.ajpath.2019.09.008

Upregulation of Fibroblast Growth Factors Caused by Heart and Neural Crest Derivatives Expressed 2 Suppression in Endometriotic Cells: A Possible Therapeutic Target in Endometriosis

若年者における子宮内膜症査読有り

中村智子

東海産婦人科内視鏡手術研究会雑誌 8 巻頁： 3 - 9 2020年

虫垂子宮内膜症により完全型虫垂重積を生じた一例

中村智子, 林祥太郎, 清水顕, 村岡彩子, 仲西菜月, 新美薫, 大須賀智子, 後藤真紀

日本産科婦人科内視鏡学会雑誌 36 巻 ( 1 ) 頁： 185 - 188 2020年

記述言語：日本語出版者・発行元：日本産科婦人科内視鏡学会

Appendiceal intussusception secondary to endometriosis is extremely rare. We report a case of left ovarian endometriosis with endometriosis-induced appendiceal intussusception in a 42-year-old woman presenting with rectal bleeding and left-sided abdominal pain. Colonoscopy revealed a cecal mass measuring 30 mm in diameter. Magnetic resonance imaging revealed extensive pelvic adhesions and a left-sided endometrioma measuring 20 mm in diameter. Laparoscopy revealed isolated appendiceal intussusception and deep infiltrating pelvic endometriosis. Laparoscopic ileocecal resection and left ovarian cystectomy were performed. Histopathological examination confirmed the diagnosis of ovarian endometriosis with endometriosis-induced appendiceal intussusception. Although rare, endometriosis-induced appendiceal intussusception should be considered in the differential diagnosis in women with endometriosis presenting with an appendiceal mass. Coordination between gynecologists and gastrointestinal surgeons is essential for an effective surgical approach to ensure optimal management.

DOI： 10.5180/jsgoe.36.1_185

CiNii Research

Kato N., Iwase A., Ishida C., Nagai T., Mori M., Bayasula , Nakamura T., Osuka S., Ganiyeva U., Qin Y., Miki R., Kikkawa F.

Reproductive Sciences 26 巻 ( 7 ) 頁： 979 - 987 2019年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Reproductive Sciences

Several features exist that distinguish endometriotic cells from eutopic endometrial cells. Progesterone resistance is one of the main distinguishing features, although how progesterone resistance affects the phenotype of endometriotic cells is not fully elucidated. Heart and neural crest derivatives expressed 2 (HAND2) is a transcriptional factor that plays an important role in maintaining endometrial function in a progesterone-dependent manner. Therefore, we explored whether progesterone-dependent HAND2 is implicated in the progression of endometriosis. HAND2 was less expressed by endometriotic tissues compared to endometrial tissues. Suppression of HAND2 expression induced fibroblast growth factor 1 (FGF1), FGF2, and FGF9 in endometriotic stromal cells and consequently enhanced migration and invasion capacity. AZD4547, a FGF receptor inhibitor, diminished the migration and invasion of endometriotic cells in vitro. In the murine model of endometriosis, AZD4547 showed suppressive effects on the development of endometriotic lesions at a relatively low concentration. In conclusion, we demonstrated that FGF1, FGF2, and FGF9 are downstream effectors of HAND2 in endometriotic cells. Since HAND2-dependent FGFs play roles in enhancing invasive capacity of endometriotic cells, our results suggest that FGF receptor inhibitors, such as AZD4547, can be promising therapeutic targets for endometriosis.

DOI： 10.1177/1933719118802053

Molecular mechanism of FSHR expression induced by BMP15 in human granulosa cells Open Access

Shimizu, K; Nakamura, T; Bayasula; Nakanishi, N; Kasahara, Y; Nagai, T; Murase, T; Osuka, S; Goto, M; Iwase, A; Kikkawa, F

JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 36 巻 ( 6 ) 頁： 1185 - 1194 2019年6月

出版者・発行元：Journal of Assisted Reproduction and Genetics

Purpose: Follicle-stimulating hormone receptor (FSHR) expression in granulosa cells is critical in enabling follicles to achieve accelerated growth. Although FSHR expression has been reported to be epigenetically regulated, the mechanism is unclear. Cooperation between oocytes and granulosa cells is also essential for normal follicular growth. Among oocyte-derived factors, bone morphogenetic protein 15 (BMP15) promotes follicular growth and is suggested to have epigenetic effects. We examined the role of BMP15 in the acquirement of FSHR in human granulosa cells. Methods: Immortalized non-luteinized human granulosa (HGrC1) cells were stimulated with trichostatin A (TSA) or BMP15 to analyze FSHR expression, histone modifications, and USF1/2 binding at the FSHR promoter region. Histone acetyl transferase (HAT) activity and phosphorylation of Smad 1/5/8 and p38 MAPK were examined with or without BMP15, SB203580, and LDN193189. CYP19A1 expression and estradiol production were also studied. Results: TSA and BMP15 induced FSHR mRNA expression in a dose-dependent manner and histone modifications were observed with increased binding of USF1/2. BMP15 increased FSHR protein expression, which was suppressed by LDN193189. BMP15 increased phosphorylation of Smad 1/5/8 and significantly increased HAT activity, which was inhibited by LDN193189, but not by SB203580. BMP15 increased phosphorylation of p38 MAPK and USF1. LDN193189 suppressed BMP15-induced phosphorylation of both p38 MAPK and USF1, whereas SB203580 suppressed the phosphorylation of USF1. BMP15 increased CYP19A1 mRNA expression and estradiol production. Conclusion: BMP15 induced FSHR expression in human granulosa cells through Smad and non-Smad pathways. This mechanism of FSHR induction by BMP15 may be utilized for controlling follicular growth.

DOI： 10.1007/s10815-019-01469-y

Animal models of polycystic ovary syndrome: A review of hormone-induced rodent models focused on hypothalamus-pituitary-ovary axis and neuropeptides Open Access

Osuka S., Nakanishi N., Murase T., Nakamura T., Goto M., Iwase A., Kikkawa F.

Reproductive Medicine and Biology 18 巻 ( 2 ) 頁： 151 - 160 2019年4月

出版者・発行元：Reproductive Medicine and Biology

Background: Polycystic ovary syndrome (PCOS) is a common endocrine disorder among women of reproductive age and a major cause of infertility; however, the pathophysiology of this syndrome is not fully understood. This can be addressed using appropriate animal models of PCOS. In this review, we describe rodent models of hormone-induced PCOS that focus on the perturbation of the hypothalamic-pituitary-ovary (HPO) axis and abnormalities in neuropeptide levels. Methods: Comparison of rodent models of hormone-induced PCOS. Main findings: The main method used to generate rodent models of PCOS was subcutaneous injection or implantation of androgens, estrogens, antiprogestin, or aromatase inhibitor. Androgens were administered to animals pre- or postnatally. Alterations in the levels of kisspeptin and related molecules have been reported in these models. Conclusion: The most appropriate model for the research objective and hypothesis should be established. Dysregulation of the HPO axis followed by elevated serum luteinizing hormone levels, hyperandrogenism, and metabolic disturbance contribute to the complex etiology of PCOS. These phenotypes of the human disease are recapitulated in hormone-induced PCOS models. Thus, evidence from animal models can help to clarify the pathophysiology of PCOS.

DOI： 10.1002/rmb2.12262

Open Access

TCF21 REGULATION OF PERIOSTIN IN THE PROGRESSION OF ENDOMETRIOTIC FIBROSIS.

Protective effects of mangafodipir against chemotherapy-induced ovarian damage in mice Open Access

Qin, Y; Iwase, A; Murase, T; Bayasula; Ishida, C; Kato, N; Nakamura, T; Osuka, S; Takikawa, S; Goto, M; Kotani, T; Kikkawa, F

REPRODUCTIVE BIOLOGY AND ENDOCRINOLOGY 16 巻 ( 1 ) 頁： 106 2018年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Reproductive Biology and Endocrinology

Background: Given the seriousness of chemotherapy-induced ovarian injury in female cancer patients, the preservation of fertility, including through the use of cryopreservation technology and pharmaceuticals, requires investigation. Previous studies have shown that damage to the ovaries is related to oxidative stress caused by anticancer drugs. Therefore, superoxide dismutase (SOD) may represent a key factor in the pharmacological protection of the ovaries. The aim of our study was to identify the effects of mangafodipir, a manganese chelate and SOD-mimetic, on suppression of apoptosis in granulosa cells and primordial follicle activation induced by anticancer drugs. Methods: Cell viability assays using methyltrichlorosilane solutions and immunoblotting for cleaved caspase-3 were performed in in vitro experiments with the simultaneous addition of mangafodipir to human non-luteinized granulosa cell line (HGrC) cultures treated with hydrogen peroxide (H2O2), cisplatin, or paclitaxel. Count and morphological analyses of follicles at each developing stage in the ovaries and immunohistochemistry for cleaved caspase-3, Ki67 and 4-hydroxynonenal, a marker for oxidative stress, were also performed using mangafodipir-injected 6-week-old female ICR mice treated with cisplatin or paclitaxel. Further, mangafodipir was injected into 6-week-old female BALB/c mice inoculated with ES-2 to analyze whether mangafodipir inhibits the anti-tumor effects of cisplatin or paclitaxel treatment. Results: Mangafodipir attenuated apoptosis induced by H2O2 and anticancer drugs in vitro. Mangafodipir also decreased the expression of 4-hydroxynonenal and reduced cisplatin- and paclitaxel-induced apoptosis in granulosa cells in vivo. In addition, mangafodipir inhibited the loss of primordial follicles. Tumor xenograft studies in mice showed that mangafodipir did not affect anticancer drug antitumor effects. Conclusions: Oxidative stress might be one of the mechanisms of cisplatin- and paclitaxel-induced the loss of primordial follicles. Mangafodipir can reduce cisplatin- and paclitaxel-induced apoptosis in granulosa cells and primordial follicle activation partially via its SOD activity. At the same time, mangafodipir might have other potential mechanisms to inhibit the activation of primordial follicles. Further, mangafodipir attenuated the ovarian damage caused by cisplatin and paclitaxel without affecting their antitumor activities. Mangafodipir, therefore, though its efficacy might be limited, may be a new option for the preservation of fertility during anticancer treatment.

DOI： 10.1186/s12958-018-0426-y

Ganieva, U; Nakamura, T; Nguyen, PX; Wei, W; Murakami, M; Miyake, N; Nakanishi, N; Kasahara, Y; Takasaki, N; Muraoka, A; Hayashi, S; Nagai, T; Murase, T; Osuka, S; Goto, M; Iwase, A

FERTILITY AND STERILITY 110 巻 ( 4 ) 頁： E393 - E393 2018年9月

Thyroid Autoantibodies do not Impair the Ovarian Reserve in Euthyroid Infertile Women: A Cross-Sectional Study

Osuka, S; Iwase, A; Goto, M; Takikawa, S; Nakamura, T; Murase, T; Kato, N; Bayasula; Kotani, T; Kikkawa, F

HORMONE AND METABOLIC RESEARCH 50 巻 ( 7 ) 頁： 537 - 542 2018年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Hormone and Metabolic Research

Patients with primary ovarian insufficiency (POI) have a high prevalence of thyroid autoimmune disorders. However, the extent of the contribution of thyroid autoantibodies or elevated thyroid-stimulating hormone (TSH) levels to decreased ovarian reserve is unclear. Therefore, we evaluated the serum levels of anti-Müllerian hormone (AMH) and thyroid autoantibodies [antithyroperoxidase antibody (TPOAb), and antithyroglobulin antibody (TgAb)] in euthyroid infertile women. One hundred and fifty-three women with normal menstrual cycles were recruited for this retrospective study. Serum levels of AMH were compared between patients with positive and negative thyroid autoantibodies. The correlation between serum levels of AMH and each thyroid autoantibody was also evaluated. Participants were observed to be either TPOAb or TgAb positive (n=27), only TPOAb positive (n=8), only TgAb positive (n=7), TPOAb and TgAb positive (double positive; n=12), and TPOAb and TgAb negative (double negative; n=126). No significant differences were found in serum AMH levels between the TPOAb- or TgAb-positive women and the antibody-double negative women. Serum AMH levels did not show a significant correlation with the concentration of TgAb or TPOAb. On the other hand, serum AMH levels negatively correlated with TSH levels in patients who were either positive for TPOAb or TgAb. Thyroid autoantibodies are not likely to influence ovarian reserve in euthyroid women whose TSH levels fall within the normal range although elevated TSH levels may be involved in the decline of serum AMH levels.

DOI： 10.1055/a-0637-9430

Neuropeptide phoenixin (PNX) and its novel receptor GPR173 induce COX-1/COX-2 expression and PGE2 production through the CREB signaling pathway

Nguyen, XP; Iwase, A; Murase, T; Ganieva, U; Qin, Y; Bayasula, B; Shimizu, K; Osuka, S; Nakamura, T; Goto, M; Kikkawa, F

HUMAN REPRODUCTION 33 巻頁： 287 - 287 2018年7月

The regulation of ovarian follicular growth by Anti-Müllerian Hormone

Nakamura T., Murase T., Osuka S., Goto M., Iwase A.

Journal of Mammalian Ova Research 35 巻 ( 1 ) 頁： 13 - 19 2018年4月

出版者・発行元：Journal of Mammalian Ova Research

Anti-Müllerian hormone (AMH) was originally discovered as the factor responsible for the regression of the Müllerian duct during male sexual differentiation. Through studies of AMH knockout mice, AMH has also been found to regulate primordial follicle recruitment and FSH-dependent cyclic recruitment. However, the details of how AMH influences follicular growth have not been elucidated. Since the early 2000s, when serum AMH concentration was found to be a reliable biochemical marker of ovarian reserve, AMH has been in the spotlight in reproductive medicine. Several studies of AMH have led to new insights on the mechanism of AMH-regulated follicular growth. Here, we review from the earliest studies to the latest findings, AMH regulation of follicle growth with reference to the potential clinical uses of AMH and AMH inhibitors.

DOI： 10.1274/jmor.35.13