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MicroRNAs secreted by the human endometrium regulates the transcriptome of the preimplantation embryo
O-439 Wednesday, October 16, 2013 05:30 PM PREGNANCY, IMPLANTATION, AND LIVE BIRTH RATES OF BLASTOCYSTS THAT WERE THAWED, ARE WARMED, BIOPSIED, VITRIFIED, AND REWARMED FOR EUPLOID BLASTOCYSTS TRANSFER. T. H. Taylor, D. G. Das, D. Whitesides, J. L. Crain, M. Wilson, J. L. Patrick. IVF, Reproductive Endocrinology Associates of Charlotte, Charlotte, NC. OBJECTIVE: Typically, blastocysts are biopsied (bx) and vitrified while awaiting preimplantation genetic screening (PGS) results. Sometimes blastocysts receive ‘‘no read’’ post PGS, leaving vitrified blastocysts with no genetic information. Furthermore, patients can desire PGS on previously frozen blastocysts which PGS was not performed. These scenarios require blastocysts to be warmed, bx’d, vitrified, and warmed again. The effect of double freezing and double warming on outcomes is unknown. DESIGN: Retrospective. MATERIALS AND METHODS: Patients undergoing blastocysts bx with PGS were included. Patients were divided into 2 groups, group 1 patients transferred (ETd) blastocysts that were vitrified and warmed once. Group 2 patients ETd blastocysts that were frozen, warmed, bx’d, vitrified, and rewarmed. RESULTS: A total of 75 patients, 36.73.9 years, and 17 patients, 35.34.9 years, were included in group 1 and 2, respectively (NS). Blastocyst warm survival rates between group 1 (101 of 103, 98.1%) and 2 (20 of 23, 87.0%) was significant (0.0420). All 75 patients received an ET from group 1, while 16 patients received an ET in group 2. The average number ET’d was 1.30.5 and 1.30.6 for group 1 and 2, respectively (NS). Chemical pregnancies (54 of 75, 72.0%) and (10 of 16, 62.5%), clinical gestations (48 of 75, 64.0%) and (9 of 16, 56.3%), clinical pregnancies (45 of 75, 60.0%) and (8 of 16, 50.0%), and implantation rates (65 of 100, 65.0%) and (11 of 20, 55.0%), were not significant between groups 1 and 2, respectively. Of the 61 clinical heartbeats from group 1, 31 were live births, 1 still birth, 25 ongoing, 3 therapeutic abortions, and 1 miscarriage. Of the 10 clinical heartbeats from group 2, 9 are live births and 1 ongoing. CONCLUSION: Although it is unconventional to thaw, bx, revitrify, and rewarm a blastocysts for ET, our results indicate that as long as the blastocyst survives the warming, outcomes are unaffected. Supported by: REACh, Charlotte, NC. O-440 Wednesday, October 16, 2013 05:45 PM ARRAY CGH AND MANDATORY SINGLE FRESH EMBRYO TRANSFER TO NORMALIZE PRETERM DELIVERY & MULTIPLE GESTATION OUTCOMES WITH IVF: FISCAL ANALYSIS OF COVERAGE EXPANSION BY A CALIFORNIA STATE HEALTH EXCHANGE. E. S. Sills,a,b G. S. Collins,c S. A. Salem,a C. A. Jones,d R. D. Salem.a aPRC-Orange County, Pacific Reproductive Center, Irvine, CA; bSchool of Life Sciences, University of Westminster, Westminster, City of London, United Kingdom; cCentre for Statistics in Medicine, Wolfson College Annexe, Oxford, Oxfordshire, United Kingdom; dGlobal Health Economics Unit, Center for Clinical & Translational Science, Burlington, VT. OBJECTIVE: To estimate economic impact of a proposed initiative entailing partial state subsidy of IVF for qualified California residents, with the proviso that patients agree to comprehensive chromosomal screening of embryos and transfer of only one embryo (SET). DESIGN: The Cardarelli formula (2000) was used to model overall economic impact of partial IVF coverage in California. MATERIALS AND METHODS: Extrapolations were based on all California non-donor IVF cycles reported in 2009 using CDC data, California vital statistics (including preterm & multiple gestation births), and published outcomes with aCGH and fresh SET. RESULTS: Total births¼526,744; Total IVF cycles initiated (completed)¼ 18,405(15,953);Births attributed to IVF¼7,546; LBW deliveries¼71,604, PT deliveries¼107,912; LBW deliveries from IVF¼4,746; PT deliveries from IVF¼4,810; Estimated term delivery cost¼$9,329; Estimated PT delivery cost¼$51,600; Cost of IVF per cycle (aCGH)¼$10,000 3,000. Accordingly, if 7,546 deliveries were attributed to IVF in 2009, and all of these births resulted from SET, then only 20.5% of 7,546 would be expected to be PT. This means that only 1,500 preterm babies would have been delivered in 2009, instead of 4,810. Avoiding 3,263 preterm births in 2009 saves California $168M in NICU spend.
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ASRM Abstracts
CONCLUSION: Our analysis suggests that even if partially subsidized IVF/aCGH/SET were available for every California IVF cycle initiated in 2009, the state would still realize a net surplus of at least $20M per year by stabilizing the IVF multiple birth rate at 3.2%. The proposal is net revenue positive for California, because although IVF/aCGH/SET is expensive, the price to obtain this technology is always less than the cost for one high-risk preterm/multiple birth. While a compelling primary interest exists to lower the multiple birth rate with IVF, this proposal also yields a socially valuable secondary benefit by improving general access to IVF for all Californians. FEMALE REPRODUCTIVE ENDOCRINOLOGY O-441 Wednesday, October 16, 2013 04:00 PM MICRORNAS SECRETED BY THE HUMAN ENDOMETRIUM REGULATES THE TRANSCRIPTOME OF THE PREIMPLANTATION EMBRYO. F. Vilella, J. M. Moreno-Moya, S. Martinez, A. Pellicer, C. Simon. IVI Investigacion, Fundacion Instituto Valenciano de Infertilidad (FIVI), Valencia University and Instituto Universitario IVI/ INCLIVA, Paterna, Valencia, Spain. OBJECTIVE: During implantation the blastocyst influences the endometrium, and the endometrial epithelium nurtures and regulates the preimplantation embryo. Our objective is to demonstrate that maternal miRNAs are transported through the endometrial fluid (EF) and uptaken by the embryo to induce modifications relevant for the establishment of pregnancy. DESIGN: To investigate the miRNA content of EF (n¼20) through the cycle identifying those differentially expressed in the window of implantation (WOI). To understand how endometrial miRNAs are secreted and taken up into the embryo where they induce adhesive modifications. MATERIALS AND METHODS: miRNAS were identified using microarrays. Exosomes were isolated by ultracentrifugation, and identified by TEM and WB. Internalization studies of miRNAs were tested with fluorescent scramble and mimic miRNAs. Embryo transcriptomic modifications were assessed by expression microarrays and adhesive modifications by embryo adhesion assay. Student’s t test was used. RESULTS: We detected 20 differentially expressed miRNAs in EF during the WOI in direct contact with the preimplantation embryo. The most represented was hsa-miR-30d. miRNAs are secreted by the endometrial epithelium both free and as exosome-associated molecules and are internalize by the embryo through the trophoectoderm. Also, miR-30d induced the overexpression of 10 genes related to cellular adhesion. Embryo adhesion was tested comparing control, scrambled miRNA, miR-30d mimic, and miR30d inhibitor (n¼360). After 32h, miR-30d mimic increased adhesion vs scramble and this was abolished with miR-30d inhibitor (35.27.4% vs 53.46.4% vs 18.53.7%, respectively p¼0.001). CONCLUSION: Our results show a model in which endometrial maternal miRNAs function as transcriptomic regulators during early embryo development, offering a new perspective on the crosstalk during implantation, and as a potential mechanism for the developmental origins of certain adult diseases such as obesity and type II diabetes. Supported by: FIVI. FV and JM contribute equally.
O-442 Wednesday, October 16, 2013 04:15 PM EVIDENCE OF GNRH ANTAGONIST ESCAPE IN OBESE WOMEN: POTENTIAL EXPLANATION FOR THE HIGH ASSISTED REPRODUCTIVE TECHNOLOGY CANCELLATION RATE. L. W. Roth,a E. L. Bradshaw-Pierce,b A. A. Allshouse,c J. Chosich,a A. J. Polotsky,a N. Santoro.a aObstetrics and Gynecology, University of Colorado, Denver, Aurora, CO; bPharmaceutical Sciences, University of Colorado, Denver, Aurora, CO; cBiostatistics and Informatics, University of Colorado, Denver, Aurora, CO. OBJECTIVE: Although obese women have an increased risk of cycle cancellation with the use of assisted reproductive technology (ART), the reasons remain unknown. LH pharmacodynamics (PD) and cetrorelix pharmacokinetics (PK) were compared in obese, ovulatory women compared with normal weight women. DESIGN: Interventional study. MATERIALS AND METHODS: A luteal phase frequent blood sampling study was done in ovulatory obese (n¼10) and normal weight women (n¼10). Protocol: 22 hours of blood draws every 10 minutes including
Vol. 100, No. 3, Supplement, September 2013
S134
ASRM Abstracts
CONCLUSION: Our analysis suggests that even if partially subsidized IVF/aCGH/SET were available for every California IVF cycle initiated in 2009, the state would still realize a net surplus of at least $20M per year by stabilizing the IVF multiple birth rate at 3.2%. The proposal is net revenue positive for California, because although IVF/aCGH/SET is expensive, the price to obtain this technology is always less than the cost for one high-risk preterm/multiple birth. While a compelling primary interest exists to lower the multiple birth rate with IVF, this proposal also yields a socially valuable secondary benefit by improving general access to IVF for all Californians. FEMALE REPRODUCTIVE ENDOCRINOLOGY O-441 Wednesday, October 16, 2013 04:00 PM MICRORNAS SECRETED BY THE HUMAN ENDOMETRIUM REGULATES THE TRANSCRIPTOME OF THE PREIMPLANTATION EMBRYO. F. Vilella, J. M. Moreno-Moya, S. Martinez, A. Pellicer, C. Simon. IVI Investigacion, Fundacion Instituto Valenciano de Infertilidad (FIVI), Valencia University and Instituto Universitario IVI/ INCLIVA, Paterna, Valencia, Spain. OBJECTIVE: During implantation the blastocyst influences the endometrium, and the endometrial epithelium nurtures and regulates the preimplantation embryo. Our objective is to demonstrate that maternal miRNAs are transported through the endometrial fluid (EF) and uptaken by the embryo to induce modifications relevant for the establishment of pregnancy. DESIGN: To investigate the miRNA content of EF (n¼20) through the cycle identifying those differentially expressed in the window of implantation (WOI). To understand how endometrial miRNAs are secreted and taken up into the embryo where they induce adhesive modifications. MATERIALS AND METHODS: miRNAS were identified using microarrays. Exosomes were isolated by ultracentrifugation, and identified by TEM and WB. Internalization studies of miRNAs were tested with fluorescent scramble and mimic miRNAs. Embryo transcriptomic modifications were assessed by expression microarrays and adhesive modifications by embryo adhesion assay. Student’s t test was used. RESULTS: We detected 20 differentially expressed miRNAs in EF during the WOI in direct contact with the preimplantation embryo. The most represented was hsa-miR-30d. miRNAs are secreted by the endometrial epithelium both free and as exosome-associated molecules and are internalize by the embryo through the trophoectoderm. Also, miR-30d induced the overexpression of 10 genes related to cellular adhesion. Embryo adhesion was tested comparing control, scrambled miRNA, miR-30d mimic, and miR30d inhibitor (n¼360). After 32h, miR-30d mimic increased adhesion vs scramble and this was abolished with miR-30d inhibitor (35.27.4% vs 53.46.4% vs 18.53.7%, respectively p¼0.001). CONCLUSION: Our results show a model in which endometrial maternal miRNAs function as transcriptomic regulators during early embryo development, offering a new perspective on the crosstalk during implantation, and as a potential mechanism for the developmental origins of certain adult diseases such as obesity and type II diabetes. Supported by: FIVI. FV and JM contribute equally.
O-442 Wednesday, October 16, 2013 04:15 PM EVIDENCE OF GNRH ANTAGONIST ESCAPE IN OBESE WOMEN: POTENTIAL EXPLANATION FOR THE HIGH ASSISTED REPRODUCTIVE TECHNOLOGY CANCELLATION RATE. L. W. Roth,a E. L. Bradshaw-Pierce,b A. A. Allshouse,c J. Chosich,a A. J. Polotsky,a N. Santoro.a aObstetrics and Gynecology, University of Colorado, Denver, Aurora, CO; bPharmaceutical Sciences, University of Colorado, Denver, Aurora, CO; cBiostatistics and Informatics, University of Colorado, Denver, Aurora, CO. OBJECTIVE: Although obese women have an increased risk of cycle cancellation with the use of assisted reproductive technology (ART), the reasons remain unknown. LH pharmacodynamics (PD) and cetrorelix pharmacokinetics (PK) were compared in obese, ovulatory women compared with normal weight women. DESIGN: Interventional study. MATERIALS AND METHODS: A luteal phase frequent blood sampling study was done in ovulatory obese (n¼10) and normal weight women (n¼10). Protocol: 22 hours of blood draws every 10 minutes including
Vol. 100, No. 3, Supplement, September 2013