Aneuploidy rates do not differ between blastocysts biopsied on day 5 (D5) versus day 6 (D6)

P-110 Tuesday, October 18, 2016 CORRECTION FACTOR REVEALS UNIFORM LEVELS OF MITOCHONDRIAL DNA IN HUMAN BLASTOCYSTS IRRESPECTIVE OF PLOIDY, AGE, OR IMPLANTATION POTENTIAL. M. Viotti, A. R. Victor, A. J. Brake, C. G. Zouves, F. L. Barnes. Zouves Fertility Center, Foster City, CA. OBJECTIVE: To develop a mathematical formula resulting in an accurate determination of mitochondrial DNA (mtDNA) levels in human blastocysts substratified by ploidy, age, and implantation potential. DESIGN: Retrospective analysis of mtDNA content in human blastocysts used in preimplantation genetic diagnosis for IVF selection. MATERIALS AND METHODS: 833 embryos derived from 181 patients were tested for mtDNA content by next generation sequencing (NGS), and 150 embryos derived from 96 patients were tested by quantitative polymerase chain reaction (qPCR). For each embryo, the level of mtDNA was determined from a trophectoderm biopsy by whole genome amplification followed by NGS and/or qPCR. The value was subjected to mathematical analysis tailored to the genomic DNA composition of said embryo. Grouped values were compared by Welch’s two-tailed upaired t-test. RESULTS: On average our quantitation method changed the conventionally determined mtDNA level of a given embryo via NGS by 1.35% +/-1.58%, with changes ranging up to 17.42%, and via qPCR by 1.33% +/-8.08%, with changes ranging up to 50.00%. Levels of mtDNA in euploid and aneuploid embryos showed a statistically insignificant difference of P¼0.102 by NGS (euploid N¼494, aneuploid N¼339) and P¼0.642 by qPCR (euploid N¼100, aneuploid N¼50). Blastocysts derived from younger or older patients had comparable mtDNA values, with P¼0.293 by NGS (20-37 age group N¼559, 38-46 age group N¼274) and P¼0.101 by qPCR (20-37 age group N¼92, 38-46 age group N¼58). Blastocysts that upon transfer resulted in implantation did not contain significantly different mtDNA levels compared to blastocysts that failed to implant, with P¼0.813 by NGS (implanted N¼51, non-implanted N¼69) and P¼0.103 by qPCR (implanted N¼49, non-implanted N¼51). CONCLUSIONS: We recommend the implementation of our correction factor to all laboratories evaluating mtDNA levels in their embryos by NGS or qPCR. Applied to our in-house data, our quantitation method reveals that overall levels of mtDNA are largely equal between human blastocysts regardless of embryo ploidy, age, or implantation potential. P-111 Tuesday, October 18, 2016 ANEUPLOIDY RATES DO NOT DIFFER BETWEEN BLASTOCYSTS BIOPSIED ON DAY 5 (D5) VERSUS DAY 6 (D6). C. M. Owen,a L. A. Bishop,b K. Koniares,c M. W. Healy,a N. Banks,a K. S. Richter,b K. Devine,d A. DeCherney,a M. J. Hill,a J. Doyle.b aPRAE, NICHD, NIH, Bethesda, MD; bShady Grove Fertility Reproductive Science Center, Rockville, MD; cGeorgetown University School of Medicine, Washington, DC; d Shady Grove Fertility Reproductive Science Center, Washington, DC. OBJECTIVE: Prior studies have demonstrated increased pregnancy rates in fresh IVF with blastocysts transferred on D5 versus D6. It has been debated whether this observed difference is due to a decrease in embryo viability with slower progression, a decline in endometrial receptivity, or a combination of both factors. Our objective was to determine if the rate of aneuploidy is increased in embryos with delayed blastulation on D6 compared to D5 in IVF cycles with PGS. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: Autologous fresh IVF cycles with preimplantation genetic screening from February 2015 to July 2015 were analyzed. Maternal age, paternal age, number of embryos biopsied, day of trophectoderm biopsy, and biopsy results were recorded for each cycle. Cy-

cles were excluded if there were known parental chromosomal abnormalities (e.g. chromosomal translocations). GEE models assessed for associations of aneuploidy status, maternal age, paternal age, and day of trophectoderm biopsy and accounted for the non-independence of day 5 and 6 biopsies within the same patient. RESULTS: 309 fresh autologous IVF cycles including 1,237 embryo biopsies (537 D5 biopsies, 700 D6 biopsies) from 267 patients were analyzed. On average, there were four embryos biopsied from each patient (1.7 biopsies/cycle on D5 and 2.2 biopsies/cycle on D6). Average maternal age of the cohort was 37.0 years and average paternal age was 39.2 years. Maternal age was significantly associated with an increased risk of aneuploidy (OR 1.02, 95% CI 1.003-1.016, P¼0.001). The adjusted aneuploidy rate between D5 and D6 biopsies was not significantly different (D5 aneuploidy 55.2% ( 36.3%) versus D6 aneuploidy 60.0% ( 37.3%); P¼0.239). CONCLUSIONS: Comparison of 1237 D5 vs. D6 blastocysts did not demonstrate a significant difference between aneuploidy rates. While it remains possible that a larger cohort could demonstrate statistical significance of the observed 5% difference in aneuploidy to be significant, these data suggest that endometrial asynchrony may play a larger role in the superior pregnancy rate seen with D5 transfer. Supported by: This research was supported, in part, by the Intramural research program of the Program in Reproductive and Adult Endocrinology, NICHD, NIH. P-112 Tuesday, October 18, 2016 THE DEGREE TO WHICH PGS INCREASES IVF EFFICIENCY IS CORRELATED WITH INCREASED MATERNAL AGE. L. Sekhon,a,b T. G. Nazem,a,b A. Zgodic,c K. Hunter Cohn,c H. Wu,d L. Grunfeld,a,b P. Yurttas Beim,c A. B. Copperman.a,b aReproductive Medicine Associates of New York, New York, NY; bObstetrics, Gynecology & Reproductive Science, Icahn School of Medicine at Mount Sinai, New York, NY; cCelmatix Inc, New York, NY; dAnalytics Team, Celmatix Inc., New York, NY. OBJECTIVE: While preimplantation genetic screening (PGS) has been shown to improve embryo implantation and reduce pregnancy loss, the age-weighted benefits have yet to be established. This study compared the number of egg retrievals, embryo transfers and length of time required to achieve pregnancy after undergoing IVF with and without PGS. DESIGN: Retrospective cohort. MATERIALS AND METHODS: Patients who underwent IVF +/- PGS (trophectoderm bx) from 2010-2015 were included. Cox proportional hazards models with restricted time to event estimates evaluated average number of fresh IVF cycles required to obtain a euploid embryo and average number of embryo transfers (ETs) until first clinical pregnancy. Time to clinical pregnancy was approximated from modeled time estimates and outcomes of fresh IVF and subsequent fresh and frozen ET cycles in non-PGS patients (following ASRM ET guidelines) and PGS patients (frozen SET (FET) used exclusively). Time estimates (days) used were: fresh IVF (21), freeze-all w/FET (45), subsequent FET (21), biochemical (31), clinical loss (91), negative pregnancy test (16), and ongoing pregnancy (42). RESULTS: A total of 10,642 IVF cycles (2,758 with PGS, 7,884 unscreened) were analyzed. Controlling for age, ovarian reserve, and embryos transferred, PGS improved clinical pregnancy rate (HR 1.4 [1.01-2.0] p ¼ 0.045) with a progressive increase with age (age 38-20: HR 2.0 [1.4-2.9], 41-42: HR 3.4 [2.0-5.8], >42: HR 7.5 [4.4-12.8], p < 0.001). Cycle outcomes are shown (Table 1). In the PGS cohort, there was an age-related increase in retrieval cycles needed to obtaining a euploid embryo. With PGS, there was a reduction in ETs required for clinical pregnancy in patients aged >39, a reduction in time to clinical pregnancy in patients aged >41, 86.6% decrease in multiples and 15.4% reduction in clinical pregnancy loss. CONCLUSIONS: The use of PGS increases the efficacy and safety of IVF in patients of all ages. In women >40, PGS markedly improves the efficiency

PGS cohort/non-PGS cohort: comparison of cycle characteristics and clinical outcome

Age

Embryos transferred (euploid/unscreened)

28 32 36 39 41 43

1/1 1/1 1/2 1/2 1/3 1/3

e148

ASRM Abstracts

Average retrievals to euploid embryo

Average transfers to clinical pregnancy

Average time to clinical pregnancy (months)

Multiple pregnancy rate (%) (observed)

Clinical loss rate (%) (observed)

1.03 / NA 1.11 / NA 1.23 / NA 1.44 / NA 1.63 / NA 1.82 / NA

1.76 / 2.27 2.12 / 2.91 2.18 / 2.14 2.2 / 3.39 1.96 / 4.29 2.01 / 6.88

4.49 / 4.42 5.46 / 5.32 5.87 / 4.14 6.76 / 6.25 6.79 / 8.15 7.42 / 11.35

2.9 / 4.1 4.3 / 2.3 0.8 / 30.1 2.8 / 19.6 3.2 / 29.6 0.0 / 8.0

5.6 / 9.0 9.9 / 10.0 8.2 / 10.0 11.3 / 12.1 6.5 / 16.5 6.2 / 11.0

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