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Estrogen and progesterone are concurrently required for uterine leiomyoma enlargement in a novel in vivo model
MATERIALS AND METHODS: A total of 215 women with otherwise unexplained primary infertility and with ultrasonographically diagnosed submucous myomas as the sole cause for fertility failure were recruited. Women were randomly allocated to one of two pretreatment groups matched by age. Hysteroscopic myomectomy was performed in the study group (n¼101). Diagnostic hysteroscopy and myoma biopsy was performed in the control group (n¼103). No fertility therapy was given for either group. Clinical pregnancy rates according to patient and myoma characteristics were the main outcome measures. RESULTS: The baseline characteristics of both patients and submucous myomas were comparable. Among patients with complete follow-up, a total of 93 (45.6%) pregnancies occured, 64 (63.4%) in the study group and 29 (28.2%) in the control. Women in the study group had a better possibility of becoming pregnant after hysteroscopic myomectomy with a relative risk of 2.1 (95% CI 1.5-2.9). No difference in pregnancy rates was observed according to size, number and location of myomas in both groups. However, fertility rates appear to increase after hysteroscopic myomectomy of type 0 and type I myomas (P<0.05). In contrast, for the subgroup of patients with type II myomas, no difference in fertility rates were noted. CONCLUSIONS: Hysteroscopic myomectomy for submucous fibroids in women with otherwise unexplained primary infertility is effective in achieving better pregnancy rate. We think that a multicenter study should be conducted before evaluating the impact of submucous myoma characteristics on fertility outcome.
O-152 Tuesday, October 20, 2009 4:45 PM OSMOTIC STRESS RESPONSE IS ALTERED IN LEIOMYOMA CELLS. D. M. McCarthy-Keith, M. Malik, J. Britten, J. Segars, W. H. Catherino. Program in Reproductive and Adult Endocrinology, NICHD, NIH, Bethesda, MD; Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, MD. OBJECTIVE: Clinical treatment of leiomyomas with GnRH analogue produces a rapid reduction in leiomyoma volume. Rapid leiomyoma regrowth when GnRH treatment is withdrawn suggests the reduction in volume is due to transport of water, yet little is known about the response of leiomyoma cells to osmotic stress. Nuclear factor of activated T cells 5 (NFAT5) is the key osmo-sensing transcription factor in human cells. Our goal was to characterize osmosensing via NFAT5 in leiomyoma cells. DESIGN: Prospective, human model experiments MATERIALS AND METHODS: Osmotic stress was induced by culturing immortalized human leiomyoma and myometrial cells in DMEM-10% FBS with additional osmolytes of NaCl at 25, 50 or 100 mM for 8 hours. Proliferation assays were performed by the sulforhodamine B method. Apoptosis was assessed by caspase 3/7 activity assay. Expression of NFAT5 in tissue and cells was determined with qRT-PCR and western blot analysis. RESULTS: Leiomyoma cell growth was inhibited 18% at 50 mM NaCl compared to 48% in myometrial cells (p<0.05), consistent with altered osmosensing in leiomyoma cells. This finding was not due to apoptosis, as evidenced by no change in caspase activity. NFAT5 transcripts were increased in leiomyoma 1.5 fold 0.19 compared to myometrium in 4 of 7 patient samples (p<0.05). Concentration dependent upregulation of NFAT5 mRNA was observed in osmotically stressed leiomyoma cells. At 50mM NaCl, NFAT5 transcripts were increased 2.5-fold compared to control. CONCLUSIONS: Leiomyoma cells were more resistant to changes in osmotic stress than myometrial cells, suggesting dysregulation in osmotic signaling in leiomyomas. This change was accompanied by differential expression of NFAT5 in leiomyoma tissue. Altered osmotic signaling in leiomyoma cells suggests a central role for water regulation in the rapid change of leiomyoma volume with GnRH analogue treatment. Supported by: This research was supported by a research grant by EMDSerono, and HD ZO1-008737-07 NIH.
O-153 Tuesday, October 20, 2009 5:00 PM ESTROGEN AND PROGESTERONE ARE CONCURRENTLY REQUIRED FOR UTERINE LEIOMYOMA ENLARGEMENT IN A NOVEL IN VIVO MODEL. H. Ishikawa, K. Ishi, R. Kakazu, S. E. Bulun, T. Kurita. Division of Reproductive Biology Research, Department of Obstetrics and Gynecology, Northwestern University, Chicago, IL; Reproductive Medicine, Graduate School of Medicine, Chiba University, Chiba, Japan.
FERTILITY & STERILITYÒ
OBJECTIVE: Although the importance of sex steroids in uterine leiomyoma is well-established, the exact function of 17b-estradiol (E) and progesterone (P) remain controversial. To elucidate the role of E and P in the growth of this tumor, we have established a novel xenograft model of human leiomyoma tissue. DESIGN: Experimental molecular study. MATERIALS AND METHODS: Surgically removed uterine leiomyoma tissue was grafted into the subrenal capsule of adult female NOD/SCID IL2Rg null (NSG) mice. At grafting, all hosts were ovariectomized and supplemented with EþP. Two weeks after grafting, hosts were divided into 4 groups and subjected to different hormone treatments (EþP, P, E, and control) for 8 weeks. Xenografts were analyzed for tumor volume, proliferation (ki67), cell density, which reflects amount of extra cellular matrix (ECM) , and expression of estrogen receptor (ER) and progesterone receptor (PR). RESULTS: All xenografts, which were subjected to EþP within the first two weeks after grafting, grew. Thereafter, only those that were exposed to EþP continued to enlarge, whereas those treated with either E or P alone shrunk significantly in size. There was no significant difference in the tumor volume between the E, P, and control groups. The ki67 index was significantly higher and cell density was significantly lower in the EþP treated group than the other 3 groups. The growth-promoting effect of EþP was completely blocked by an anti-progestin, RU486. ERa was expressed in the xenografts of all groups. In contrast, expression of PR was totally dependent on exposure to E. CONCLUSIONS: Our xenograft model faithfully preserves the phenotype and hormone-responsiveness of human leiomyomata in situ. The growth and maintenance of human uterine leiomyomata were dependent on both E and P. P action via PR increases tumor volume through cell proliferation and ECM accumulation ,while E is required to sensitize cells to P by inducing PR. Supported by: NIH-HD46260, Friends of Prentice.
O-154 Tuesday, October 20, 2009 5:15 PM UNDERSTANDING CELLULAR LEIOMYOMAS: A CASE CONTROL STUDY. F. A. Taran, A. L. Weaver, B. S. Gostout, E. A. Stewart. Department of Obstetrics and Gynecology, Mayo Clinic, Rochester, MN; Department of Obstetrics and Gynecology, Medical Faculty of Otto-von-Guericke University, Magdeburg, Sachsen-Anhalt, Germany; Department of Health Sciences Research, Mayo Clinic, Rochester, MN. OBJECTIVE: Rare reports suggest that cellular leiomyomas have a phenotype more consistent with malignancy that conventional leiomyomas. The purpose of this study is to examine this rare lesion to determine if cellular leiomyomas have a unique clinical presentation compared to typical leiomyomas. DESIGN: Retrospective case-control study. MATERIALS AND METHODS: This study was conducted at the Mayo Clinic, Rochester, MN. Women undergoing surgical procedures between January 1, 1989 and December 31, 2008 with a histologic diagnosis of uterine cellular leiomyomas comprised the study group (N¼99). Control subjects were women undergoing the same procedure by the same surgeon within 2 years with a diagnosis of leiomyoma. Control subjects were matched in a 2:1 ratio to study subjects (N¼ 198). A review of hospital and ambulatory records was performed to ascertain sociodemographic and anthropometric variables as well as intraoperative and pathologic findings. RESULTS: In 99% of the uterine specimens with multiple leiomyomas, the cellular leiomyoma was the largest mass. A single uterine mass was more common among the cellular leiomyoma cases compared to women with typical leiomyomas (OR 2.1, 95% CI 1.3-3.4). Women with cellular leiomyomas were significantly younger (P¼.010) and had uteri with decreased weight on pathologic exam (P<.0001) compared to typical fibroid uteri. Additionally, more women with cellular leiomyomas had surgery for leiomyoma(s) or leiomyoma related-symptoms (P¼.003) than women with typical leiomyomas. In multivariable logistic regression analyses, women with cellular leiomyomas were more likely to have surgical indication for enlarging leiomyoma (OR 7.1, CI 2.4-25.1), were more likely to have more fibroid burden (OR per doubling in fibroid size 1.2, CI 1.1-1.3) and have fewer leiomyomas (OR 0.9, CI 0.9-1.0) when compared to women with typical leiomyomas. CONCLUSIONS: Cellular leiomyomas have a distinct clinical phenotype compared to typical leiomyomas and some characteristics common with leiomyosarcomas.
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O-152 Tuesday, October 20, 2009 4:45 PM OSMOTIC STRESS RESPONSE IS ALTERED IN LEIOMYOMA CELLS. D. M. McCarthy-Keith, M. Malik, J. Britten, J. Segars, W. H. Catherino. Program in Reproductive and Adult Endocrinology, NICHD, NIH, Bethesda, MD; Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, MD. OBJECTIVE: Clinical treatment of leiomyomas with GnRH analogue produces a rapid reduction in leiomyoma volume. Rapid leiomyoma regrowth when GnRH treatment is withdrawn suggests the reduction in volume is due to transport of water, yet little is known about the response of leiomyoma cells to osmotic stress. Nuclear factor of activated T cells 5 (NFAT5) is the key osmo-sensing transcription factor in human cells. Our goal was to characterize osmosensing via NFAT5 in leiomyoma cells. DESIGN: Prospective, human model experiments MATERIALS AND METHODS: Osmotic stress was induced by culturing immortalized human leiomyoma and myometrial cells in DMEM-10% FBS with additional osmolytes of NaCl at 25, 50 or 100 mM for 8 hours. Proliferation assays were performed by the sulforhodamine B method. Apoptosis was assessed by caspase 3/7 activity assay. Expression of NFAT5 in tissue and cells was determined with qRT-PCR and western blot analysis. RESULTS: Leiomyoma cell growth was inhibited 18% at 50 mM NaCl compared to 48% in myometrial cells (p<0.05), consistent with altered osmosensing in leiomyoma cells. This finding was not due to apoptosis, as evidenced by no change in caspase activity. NFAT5 transcripts were increased in leiomyoma 1.5 fold 0.19 compared to myometrium in 4 of 7 patient samples (p<0.05). Concentration dependent upregulation of NFAT5 mRNA was observed in osmotically stressed leiomyoma cells. At 50mM NaCl, NFAT5 transcripts were increased 2.5-fold compared to control. CONCLUSIONS: Leiomyoma cells were more resistant to changes in osmotic stress than myometrial cells, suggesting dysregulation in osmotic signaling in leiomyomas. This change was accompanied by differential expression of NFAT5 in leiomyoma tissue. Altered osmotic signaling in leiomyoma cells suggests a central role for water regulation in the rapid change of leiomyoma volume with GnRH analogue treatment. Supported by: This research was supported by a research grant by EMDSerono, and HD ZO1-008737-07 NIH.
O-153 Tuesday, October 20, 2009 5:00 PM ESTROGEN AND PROGESTERONE ARE CONCURRENTLY REQUIRED FOR UTERINE LEIOMYOMA ENLARGEMENT IN A NOVEL IN VIVO MODEL. H. Ishikawa, K. Ishi, R. Kakazu, S. E. Bulun, T. Kurita. Division of Reproductive Biology Research, Department of Obstetrics and Gynecology, Northwestern University, Chicago, IL; Reproductive Medicine, Graduate School of Medicine, Chiba University, Chiba, Japan.
FERTILITY & STERILITYÒ
OBJECTIVE: Although the importance of sex steroids in uterine leiomyoma is well-established, the exact function of 17b-estradiol (E) and progesterone (P) remain controversial. To elucidate the role of E and P in the growth of this tumor, we have established a novel xenograft model of human leiomyoma tissue. DESIGN: Experimental molecular study. MATERIALS AND METHODS: Surgically removed uterine leiomyoma tissue was grafted into the subrenal capsule of adult female NOD/SCID IL2Rg null (NSG) mice. At grafting, all hosts were ovariectomized and supplemented with EþP. Two weeks after grafting, hosts were divided into 4 groups and subjected to different hormone treatments (EþP, P, E, and control) for 8 weeks. Xenografts were analyzed for tumor volume, proliferation (ki67), cell density, which reflects amount of extra cellular matrix (ECM) , and expression of estrogen receptor (ER) and progesterone receptor (PR). RESULTS: All xenografts, which were subjected to EþP within the first two weeks after grafting, grew. Thereafter, only those that were exposed to EþP continued to enlarge, whereas those treated with either E or P alone shrunk significantly in size. There was no significant difference in the tumor volume between the E, P, and control groups. The ki67 index was significantly higher and cell density was significantly lower in the EþP treated group than the other 3 groups. The growth-promoting effect of EþP was completely blocked by an anti-progestin, RU486. ERa was expressed in the xenografts of all groups. In contrast, expression of PR was totally dependent on exposure to E. CONCLUSIONS: Our xenograft model faithfully preserves the phenotype and hormone-responsiveness of human leiomyomata in situ. The growth and maintenance of human uterine leiomyomata were dependent on both E and P. P action via PR increases tumor volume through cell proliferation and ECM accumulation ,while E is required to sensitize cells to P by inducing PR. Supported by: NIH-HD46260, Friends of Prentice.
O-154 Tuesday, October 20, 2009 5:15 PM UNDERSTANDING CELLULAR LEIOMYOMAS: A CASE CONTROL STUDY. F. A. Taran, A. L. Weaver, B. S. Gostout, E. A. Stewart. Department of Obstetrics and Gynecology, Mayo Clinic, Rochester, MN; Department of Obstetrics and Gynecology, Medical Faculty of Otto-von-Guericke University, Magdeburg, Sachsen-Anhalt, Germany; Department of Health Sciences Research, Mayo Clinic, Rochester, MN. OBJECTIVE: Rare reports suggest that cellular leiomyomas have a phenotype more consistent with malignancy that conventional leiomyomas. The purpose of this study is to examine this rare lesion to determine if cellular leiomyomas have a unique clinical presentation compared to typical leiomyomas. DESIGN: Retrospective case-control study. MATERIALS AND METHODS: This study was conducted at the Mayo Clinic, Rochester, MN. Women undergoing surgical procedures between January 1, 1989 and December 31, 2008 with a histologic diagnosis of uterine cellular leiomyomas comprised the study group (N¼99). Control subjects were women undergoing the same procedure by the same surgeon within 2 years with a diagnosis of leiomyoma. Control subjects were matched in a 2:1 ratio to study subjects (N¼ 198). A review of hospital and ambulatory records was performed to ascertain sociodemographic and anthropometric variables as well as intraoperative and pathologic findings. RESULTS: In 99% of the uterine specimens with multiple leiomyomas, the cellular leiomyoma was the largest mass. A single uterine mass was more common among the cellular leiomyoma cases compared to women with typical leiomyomas (OR 2.1, 95% CI 1.3-3.4). Women with cellular leiomyomas were significantly younger (P¼.010) and had uteri with decreased weight on pathologic exam (P<.0001) compared to typical fibroid uteri. Additionally, more women with cellular leiomyomas had surgery for leiomyoma(s) or leiomyoma related-symptoms (P¼.003) than women with typical leiomyomas. In multivariable logistic regression analyses, women with cellular leiomyomas were more likely to have surgical indication for enlarging leiomyoma (OR 7.1, CI 2.4-25.1), were more likely to have more fibroid burden (OR per doubling in fibroid size 1.2, CI 1.1-1.3) and have fewer leiomyomas (OR 0.9, CI 0.9-1.0) when compared to women with typical leiomyomas. CONCLUSIONS: Cellular leiomyomas have a distinct clinical phenotype compared to typical leiomyomas and some characteristics common with leiomyosarcomas.
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