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The ovarian steroid and prostaglandin may induce alterations of microrna expression in endometrial stromal cell
P-186 Tuesday, October 20, 2015 USE OF CLOMIPHENE CITRATE IN MINIMAL OVARIAN STIMULATION CYCLES NEGATIVELY IMPACTS ENDOMETRIAL THICKNESS, INDEPENDENT OF PEAK ESTRADIOL LEVELS: AN ARGUMENT FOR A FREEZE-ALL APPROACH. B. G. Reed, M. Ezzati, S. N. Babayev, V. Libby, B. Carr, O. Bukulmez. UT Southwestern Medical Center, Dallas, TX. OBJECTIVE: Minimal ovarian stimulation cycles use daily clomiphene citrate (CC) and a small amount of gonadotropin on days 5, 7, and 9 of the ovarian stimulation. Extrapolating from data on intrauterine insemination cycles that indicate CC may have a detrimental effect on the endometrial stripe (ES), we avoid fresh embryo transfer in patients undergoing minimal stimulation despite the lack of data from IVF cycles to support this freeze-all practice. We sought to determine if ES is negatively affected in minimal stimulation as compared to other types of ovarian stimulation that do not utilize CC. In addition, we sought to compare each patient’s ES during her minimal stimulation cycle(s) with her ES during her subsequent frozen embryo transfer (FET) cycle. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: A total cohort of 141 cycles (75 patients) were analyzed: 78 minimal ovarian stimulation cycles and their 31 subsequent FETs, 13 mild stimulation cycles, and 19 high dose gonadotropin GnRH antagonist cycles. One way ANOVA and Tukey’s test were used for continuous variables. Chi square test was used to compare the cumulative live birth rate per patient. Paired t-test was used to compare the ES in the same patient during the minimal stimulation and her subsequent FET cycle. RESULTS: There was no statistically significant difference in age, body mass index (BMI), antimullerian hormone (AMH) level, or cumulative live birth rates between the study groups (Table 1). Maximum ES in the minimal stimulation group was significantly lower than the other two groups (8.1 mm versus 13.6 mm and 13.9 mm). Peak estradiol level was significantly higher in the high dose gonadotropin group as compared to both the minimal or mild stimulation groups. Despite a significant difference in peak estradiol levels, there was no difference in ES between the mild and high-dose stimulation groups. In patients who underwent minimal stimulation IVF followed by FET, significantly thicker ES was observed during their FET cycles (7.9 mm vs 10.3 mm). CONCLUSIONS: Based on our data, endometrial thickness is negatively impacted during minimal stimulation IVF cycles, independent of peak estradiol levels. The negative effects on the endometrial thickness are not observed in the subsequent FET cycles and, hence, a freeze-all approach can be adopted to mitigate potential adverse endometrial effects in minimal stimulation IVF cycles. Table 1. Demographics and outcomes. High dose Frozen embryo Mild gonadotropin transfer following Minimal stimulation GnRH antag minimal stimulation (Did not stimulation stimulation (Included use include (Did not include (Did not include of CC) the use of CC) the use of CC) the use of CC) p-value Age (y) 384 38.83.5 BMI 264.8 25.65.7 AMH (ng/ml) 0.60.7 0.50.5 Maximum ES (mm) 8.12 13.63.5 Peak estradiol (pg/ml) 1228730 976.9257 Paired (between 7.91.8 minimal stimulation patients and their subsequent FET) maximum ES (mm) Percent that *No fresh 33% achieved a live transfers birth per patient performedsee the FET column
37.64.5 25.24.1 1.00.7 13.93.8 24481116 -
10.31.9
NS NS NS <0.01 <0.01 <0.0001
19%
39%
NS
Note: Values are expressed as either percentage or meanSD.
P-187 Tuesday, October 20, 2015 THE OVARIAN STEROID AND PROSTAGLANDIN MAY INDUCE ALTERATIONS OF MICRORNA EXPRESSION IN ENDOMETRIAL STROMAL CELL. S. Oh, M. Han, Y. Cho, J. Bae, J. Park, S. Park. Obstetrics and Gynecology, Dong-A University Hospital, Busan, Korea, Republic of.
e170
ASRM Abstracts
OBJECTIVE: microRNAs (miRNAs) are known to play a role in regulation of genes related to the progression of pathological endometrium. Several differentially expressed miRNAs are involved in proliferation and inflammation of endometrial stromal cells (ESCs). These miRNAs are probably influenced by hormonal alteration or inflammatory reaction. We performed this experiment with selected eight miRNAs to verify the hypothesis that ovarian steroids or inflammatory substances cause regulatory effects on miRNAs expression. These miRNAs are known to involved in cell proliferation (Let 7a, miR-21, miR-26a, and miR-200a) or inflammation reaction (miR-29b, miR-93, miR-106b, and miR-100b). DESIGN: Experimental study. MATERIALS AND METHODS: Total RNA was extracted from endometrial stromal cells (ESCs). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to determine the levels of miRNA. ESCs were treated with Estradiol (E2), Progesterone (P4), Prostaglandin E2 (PGE2), Prostaglandin F2a (PGF2a), E2+fulvestrant (ICI), P4+mifepriston (MF) and PGE2+celecoxib (Cele). RESULTS: According to qRT-PCR, in E2-treated ESCs, expression levels of Let 7a, miR-21 and miR-26a related to proliferation were statistically significantly lower than in controls (P<0.05). Also, in E2-treated ESCs, expression levels of miR-93 related to inflammation were statistically significantly inhibited (P<0.05). While, in P4-treated ESCs, Let 7a and miR-200a related to proliferation were statistically significantly highly expressed (P<0.05). As to PGE2-treated ESCs, expression levels of miR-21 and miR-200 related to proliferation showed a trend toward down-regulation (P<0.05). Also, in PGE2-treated ESCs, expression levels of miR-93 related to inflammation were inhibited (P<0.05). CONCLUSIONS: In present study, we identified the alterations of miRNAs expression in ESCs with ovarian hormones and inflammatory substances, leading to ensure that these consecutive processes involved in proliferation and inflammation in pathologic endometrium. Further, understanding associated with several differentially expressed miRNAs may provide useful information about the pathogenesis of gynecologic diseases.
P-188 Tuesday, October 20, 2015 DECIDUALIZED ENDOMETRIAL STROMAL CELLS ARE CAPABLE OF DRIVING M2 MACROPHAGE DIFFERENTIATION. J. Huang P. Kuo. Obstetrics and Gynecology, Division of Genetics, Tainan, Taiwan. OBJECTIVE: During decidualization, the recruited monocytes in the endometrium would undergo M2 macrophage differentiation with an immunosuppressive phenotype. Inappropriate macrophage differentiation is associated with miscarriage and preeclampsia. However, the regulations of the macrophages differentiation in the endometrium is unclear. In this study, we want to explore whether the macrophage differentiation could be regulated by decidualized endometrial stromal cells. DESIGN: hTERT-immortalized human endometrial stromal cells (THESCs) were treated with medroxyprogesterone acetate (MPA) and 8bromo-cAMP to induce decidualization. Human monocytic cell line THP1 cells primed with phorbol 12-myristate 13-acetate (PMA) or peripheral blood monocytes were cultured in the conditioned medium of decidualized T-HESCs (D(+)CM) or non-decidualized T-HESCs (D(-)CM) to induce macrophage differentiation. MATERIALS AND METHODS: The macrophage differentiation of PMA-primed THP-1 (pTHP-1) or monocytes induced by D(+)CM or D(-) CM were evaluated by detecting the expressions of CD markers and cytokines using flowcytometry and ELISA, respectively. The effects of the CM-treated pTHP-1 on the decidualization of T-HESCs and the invasion of HTR-8 trophoblast cells were evaluated. The differences between groups were analyzed using the Student t test. RESULTS: D(+)CM-treated pTHP-1 or D(+)CM-treated monocytes had a M2 differentiation with downregulations of CD11c, IL-1b, and TNF-a, and upregulations of CD163, IL-10, and TGF-b, compared with that of D(-)CMtreated cells. D(+)CM-treated pTHP-1 could maintain the decidualization of T-HESCs and promote the cell invasion of HTR-8 compared with D(-)CMtreated pTHP-1. CONCLUSIONS: The secretion of decidualized endometrial stromal cells could induce a M2 macrophage differentiation that may help the maintenance of decidualization and the development of placenta in vivo. Supported by: This research was Supported by grants from the National Science Council of the Republic of China.
Vol. 104, No. 3, Supplement, September 2015
37.64.5 25.24.1 1.00.7 13.93.8 24481116 -
10.31.9
NS NS NS <0.01 <0.01 <0.0001
19%
39%
NS
Note: Values are expressed as either percentage or meanSD.
P-187 Tuesday, October 20, 2015 THE OVARIAN STEROID AND PROSTAGLANDIN MAY INDUCE ALTERATIONS OF MICRORNA EXPRESSION IN ENDOMETRIAL STROMAL CELL. S. Oh, M. Han, Y. Cho, J. Bae, J. Park, S. Park. Obstetrics and Gynecology, Dong-A University Hospital, Busan, Korea, Republic of.
e170
ASRM Abstracts
OBJECTIVE: microRNAs (miRNAs) are known to play a role in regulation of genes related to the progression of pathological endometrium. Several differentially expressed miRNAs are involved in proliferation and inflammation of endometrial stromal cells (ESCs). These miRNAs are probably influenced by hormonal alteration or inflammatory reaction. We performed this experiment with selected eight miRNAs to verify the hypothesis that ovarian steroids or inflammatory substances cause regulatory effects on miRNAs expression. These miRNAs are known to involved in cell proliferation (Let 7a, miR-21, miR-26a, and miR-200a) or inflammation reaction (miR-29b, miR-93, miR-106b, and miR-100b). DESIGN: Experimental study. MATERIALS AND METHODS: Total RNA was extracted from endometrial stromal cells (ESCs). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to determine the levels of miRNA. ESCs were treated with Estradiol (E2), Progesterone (P4), Prostaglandin E2 (PGE2), Prostaglandin F2a (PGF2a), E2+fulvestrant (ICI), P4+mifepriston (MF) and PGE2+celecoxib (Cele). RESULTS: According to qRT-PCR, in E2-treated ESCs, expression levels of Let 7a, miR-21 and miR-26a related to proliferation were statistically significantly lower than in controls (P<0.05). Also, in E2-treated ESCs, expression levels of miR-93 related to inflammation were statistically significantly inhibited (P<0.05). While, in P4-treated ESCs, Let 7a and miR-200a related to proliferation were statistically significantly highly expressed (P<0.05). As to PGE2-treated ESCs, expression levels of miR-21 and miR-200 related to proliferation showed a trend toward down-regulation (P<0.05). Also, in PGE2-treated ESCs, expression levels of miR-93 related to inflammation were inhibited (P<0.05). CONCLUSIONS: In present study, we identified the alterations of miRNAs expression in ESCs with ovarian hormones and inflammatory substances, leading to ensure that these consecutive processes involved in proliferation and inflammation in pathologic endometrium. Further, understanding associated with several differentially expressed miRNAs may provide useful information about the pathogenesis of gynecologic diseases.
P-188 Tuesday, October 20, 2015 DECIDUALIZED ENDOMETRIAL STROMAL CELLS ARE CAPABLE OF DRIVING M2 MACROPHAGE DIFFERENTIATION. J. Huang P. Kuo. Obstetrics and Gynecology, Division of Genetics, Tainan, Taiwan. OBJECTIVE: During decidualization, the recruited monocytes in the endometrium would undergo M2 macrophage differentiation with an immunosuppressive phenotype. Inappropriate macrophage differentiation is associated with miscarriage and preeclampsia. However, the regulations of the macrophages differentiation in the endometrium is unclear. In this study, we want to explore whether the macrophage differentiation could be regulated by decidualized endometrial stromal cells. DESIGN: hTERT-immortalized human endometrial stromal cells (THESCs) were treated with medroxyprogesterone acetate (MPA) and 8bromo-cAMP to induce decidualization. Human monocytic cell line THP1 cells primed with phorbol 12-myristate 13-acetate (PMA) or peripheral blood monocytes were cultured in the conditioned medium of decidualized T-HESCs (D(+)CM) or non-decidualized T-HESCs (D(-)CM) to induce macrophage differentiation. MATERIALS AND METHODS: The macrophage differentiation of PMA-primed THP-1 (pTHP-1) or monocytes induced by D(+)CM or D(-) CM were evaluated by detecting the expressions of CD markers and cytokines using flowcytometry and ELISA, respectively. The effects of the CM-treated pTHP-1 on the decidualization of T-HESCs and the invasion of HTR-8 trophoblast cells were evaluated. The differences between groups were analyzed using the Student t test. RESULTS: D(+)CM-treated pTHP-1 or D(+)CM-treated monocytes had a M2 differentiation with downregulations of CD11c, IL-1b, and TNF-a, and upregulations of CD163, IL-10, and TGF-b, compared with that of D(-)CMtreated cells. D(+)CM-treated pTHP-1 could maintain the decidualization of T-HESCs and promote the cell invasion of HTR-8 compared with D(-)CMtreated pTHP-1. CONCLUSIONS: The secretion of decidualized endometrial stromal cells could induce a M2 macrophage differentiation that may help the maintenance of decidualization and the development of placenta in vivo. Supported by: This research was Supported by grants from the National Science Council of the Republic of China.
Vol. 104, No. 3, Supplement, September 2015