Histone deacetylases in eutopic and ectopic endometrial tissue of patients with endometriosis

of epithelial and stromal cells in the implants. Par treatment attenuated these LPS-induced stimulatory effects. CONCLUSIONS: LPS-induced pelvic inflammation status enhanced the development of murine endometriosis-like lesions via NF-kB pathway. Supported by: KAKENHI (Japan Society for the Promotion of Science Grant-in-Aid). P-442 Wednesday, October 19, 2016 HISTONE DEACETYLASES IN EUTOPIC AND ECTOPIC ENDOMETRIAL TISSUE OF PATIENTS WITH ENDOMETRIOSIS. C. Kim, J. Park, Y. Jeung, J. Moon. Obstetrics and Gynecology, College of Medicine, University of Ulsan, Asan Medical Center, Seoul, Korea, Republic of. OBJECTIVE: We performed this study to investigate the expression of class 1, 2, 3, 4 and 5 histone deacetylase (HDAC-1, 2, 3, 4, and 5) mRNA and proteins in eutopic and ectopic endometrial tissues of patients with endometriosis and in eutopic endometrial tissues of women without endometriosis, and to evaluate the relationship between HDAC and the development of endometriosis. DESIGN: Prospective cohort study. MATERIALS AND METHODS: For this prospective cohort study, twenty patients who underwent surgery for stage III or IV endometriosis for the study group and 20 patients who underwent surgery for other benign gynecologic disease for controls between Jul 2010 to Mar 2013. All subjects were not pregnant and had normal regular menstruation. No one received hormonal therapy for at least 6 months before surgical treatment. During surgical treatment, eutopic endometrial tissues were collected from study and control groups and ectopic endometrial tissues were collected from study group. To quantify the expression of HDAC mRNA, real time reverse transcriptase polymerase chain reaction (RTPCR) was employed. To measure the expression of HDAC protein, Western blotting was employed. Mean values were expressed as mean
standard deviation (SD). Kruskal-Wallis test was used to compare the mean value. Statistical significance was defined as P<0.05. All analyses were performed by using SPSS statistical package for Windows, version 11.0 (SPSS Inc, Chicago, IL). RESULTS: The amounts of HDAC-1, 2, 3, 4, and 5 mRNAs were significantly higher in eutopic and ectopic endometrial tissue of patients with endometriosis compared to eutopic endometrial tissue of normal women (p < 0.05). The expression of HDAC 1 protein was significantly higher in eutopic and ectopic endometrial tissue of patients with endometriosis. CONCLUSIONS: Our results suggest the possible relationship between the development ofendometriosis and HDAC. Especially high level of HDAC 1 in endometrial tissue may be associated with the development of endometriosis. References: 1. Eleftherios P. Samartzis, MD1, Aurelia Noske, et al. The Expression of histone deacetylase 1, but not other class I histone deacetylases, is significantly increased in endometriosis. Reprod Sci 2013;20(12):1416-22.

P-443 Wednesday, October 19, 2016 SYSTEMATIC LITERATURE REVIEW AND META-ANALYSIS REVEALS THE BIOLOGICAL FUNCTIONS MOST FREQUENTLY ALTERED IN ENDOMETRIOSIS. D. Parfitt, R. Berro, T. Hu-Seliger, A. Oromendia, C. Clementi, K. Kalmbach, S. Arunajadai, P. Yurttas Beim. Celmatix Inc, New York, NY. OBJECTIVE: As the price of genetic sequencing has plummeted over the last decade, the evidence base for genetic drivers of reproductive disorders has grown. One area of focus in particular for genetic studies has been endometriosis, which impacts 10% of all women. Here we undertook a systematic literature review and meta-analysis of human genetic studies in women with endometriosis in order to better understand the biological pathways that are commonly impacted. DESIGN: Systematic literature review and meta-analysis. MATERIALS AND METHODS: From the 25 million citations within the NCBI Pubmed repository, our natural language processing algorithms

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ASRM Abstracts

identified 4,516 reports on the molecular and genetic drivers of endometriosis. 868 of these articles (which included 893 distinct case-control experiments) reported a statistical or functional association between one or more genetic region(s) and endometriosis. We assessed these associations using the clinical validity classification system, defined by the American College of Medical Genetics (ACMG), to rank these genes according to the strength of their association with endometriosis. We then utilized a custom, fertilitycentric genome annotation database to categorize the biological functions associated with these genetic regions. RESULTS: We found that 91% of the endometriosis cases included in our review were confirmed surgically, 65% of these by laparotomy. Of the papers reviewed, 40 genes showed adequate evidence to be categorized as having ‘‘strong’’ evidence of a causal role in endometriosis according to ACMG ranking guidelines. Furthermore, we observed that these genes have well established roles in 1) the immune response, 2) tissue remodeling, 3) cell proliferation and differentiation, and 4) DNA replication and repair. CONCLUSIONS: It is thought that the number of confirmed endometriosis cases is significantly lower than the true incidence of the disease, owing in part to lack of non-invasive diagnostic tools. Our study shows that the evidence base for genetic markers of endometriosis has finally reached the same level as for many markers that are now commonly used in other fields of medicine, such as oncology, for identifying people at significantly elevated risk for developing the disease. As with cancer, early intervention with endometriosis is key to minimizing long term impacts, including infertility. Leveraging these biomarkers to help identify women who could benefit from early intervention could help minimize the long-term impacts of allowing endometriosis to go undiagnosed and untreated. P-444 Wednesday, October 19, 2016 DOES MEDICAL THERAPY EFFECT CIRCULATING MIRNAS? E. Cosar,a R. Mamillapalli,b I. Moridi,c A. Duleba,d aa b H. S. Taylor. Yale School of Medicine, New Haven, CT; Obstretics, Gynecology and Reproductive Sciences, Research, New Haven, CT; cObstetrics and Gynecology, Infertility, New Haven, CT; dReproductive Medicine, University of California, San Diego, La Jolla, CA. OBJECTIVE: We have previously identified circulating microRNAs (miRs) that can serve as biomarkers of endometriosis. Altered levels of several miRs are highly correlated with the presence of endometriosis in women. Here we determined prospectivly that endometriosis leads to alterations iin these biomarkers and that levels of circulasting miRs are altered by medical therapy in a controlled non-human primate model. DESIGN: Statins have been previously shown to decreased endometriosis in animals and humans. Experimental endometriosis was induced in sixteen baboons and half were randomized to treatment with Simvastatin. At 0 and 6 weeks all underwent serum MiR analysis and laparoscopy. MATERIALS AND METHODS: Before each surgery serum was obtained and total RNA was extracted and Poly(A) Reverse Transcription-Ploymerase Chain reaction was conducted with Invitrogen NCode miRNA First-Strand cDNA profile Synthesis MIRC-50 kit (Invitrogen). Conventional RT-qPCR was performed to detect differences between the groups. The relative expression of each miRNA was normalized by U6. Mann-Whitney U test was used to compare differences between groups. RESULTS: Before induction of endometriosis there were no statistically significant difference in serum miR levels between groups. At 6 weeks miR-125b-5p, miR-150-5p were significantly increased in the untreated endometriosis group (p¼0.02, p¼0.03 respectively). miR-3613-5p was significantly increased in the untreated endometriosis group (p¼0.04). Each of these alterations was decreased in animals treated with Simvastatin as shown in Table 1. CONCLUSIONS: We demonstrate differential expression of miRNAs in the serum of baboons after creation of experimental endometriosis.

microRNA levels in treated and untreated animals.

miR-125b-5p miR-150-5p miR-3613-5p

Endometriosis+Simvastatin

Endometriosis

P

0.029
0.018 0.033
0.021 0.551
0.108

0.116
0.055 0.189
0.062 0.386
0.087

0.02 0.03 0.04

Vol. 106, No. 3, Supplement, September 2016