Selection of single euploid blastocysts for transfer via preimplantation genetic screening with either next-generation sequencing or array comparative genomic hybridization: a randomized pilot study

MATERIALS AND METHODS: Patients with normal ovarian reserve % 42 years of age were recruited. Following routine IVF care, trophectoderm biopsy was performed. Embryos were subsequently selected for transfer per routine. No NGS CCS analysis was done prior to transfer. A novel targeted amplification method of NGS based CCS which does not require whole genome amplification was then performed. The outcome for each transferred embryo was compared to the NGS screening result to determine the predictive value of that result. In the case of a two embryo transfer, DNA fingerprinting was utilized to ensure that the embryo responsible for the pregnancy was identified correctly. Implantation rates were compared between embryos designated euploid, aneuploid, and for the population as a whole. RESULTS: 117 patients had 187 blastocysts transferred. Of the 41 embryos assigned an aneuploid karyotype, none sustained implantation yielding a predictive value of an aneuploid result of 100%. 97 of 146 embryos designated as euploid implanted and progressed to delivery yielding a predictive value for a euploid result of 66.0%. No embryo designated as euploid subsequently developed into an aneuploid gestation. Euploid embryos had a higher sustained implantation rate than the population as a whole (50.8%, p<0.0001) and those designated aneuploid (0%, p<0.0001). CONCLUSIONS: This study represents the first prospective clinical evaluation of the predictive values of both a euploid and an aneuploid result when using NGS based CCS. This targeted amplification NGS CCS paradigm has a high degree of precision and provides excellent predictive values for actual clinical outcomes. Supported by: Ferring O-32 Monday, October 19, 2015 11:30 AM PERINATAL OUTCOMES AMONG IVF SINGLETONS ARE REASSURING AFTER COMPREHENSIVE CHROMOSOMAL SCREENING AND FROZEN BLASTOCYST TRANSFER. L. A. Kondapalli, R. Collins, W. B. Schoolcraft, M. KatzJaffe. Colorado Center for Reproductive Medicine, Lone Tree, CO. OBJECTIVE: Comprehensive chromosomal screening (CCS) allows for selection of euploid blastocysts with high implantation and live birth rates. However little in known about the impact of trophectoderm biopsy on the developmental future of resultant pregnancies. The objective of this study was to compare perinatal outcomes of singleton infants conceived after frozen embryo transfer (FET) with CCS and without CCS. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: Non-smoking women who underwent autologous FET cycles from 2013-2014 were stratified by CCS (n¼139) or non-CCS (n¼308) exposure status. All embryos were biopsied at the blastocyst stage on either day 5 or 6 prior to vitrification. Standard protocols were used for hormone supplementation prior to FET. IVF cycle and birth outcomes were analyzed using appropriate parametric and non-parametric comparisons with STATA 13.1 (Stata Corp; College Station, TX, USA) with P value <0.05 considered significant. Cycles resulting in a multiple gestation were excluded. RESULTS: Although women in the CCS group were older than in the nonCCS group, mean FSH, gravidity, and history of prior ART cycles were comparable between the groups [Table 1]. Further, implantation rate is higher in the CCS group despite fewer number of embryos transferred. The majority of deliveries occurred at term (39.0 weeks) and there was no difference in infant gender between groups (49.6 vs. 46.9% female, ns). Mean gestational age and birth weight were comparable between the groups. In fact, the incidence of very preterm birth and large for gestational age was reduced in the CCS group (ns).. Cycle Characteristics and Birth Outcomes

Cycle age (y) Primigravid (n,%) BMI (kg/m2) Max FSH (mIU/ml) # embryos transferred Implantation rate (%) Gestational age (wks) Preterm birth <37wks Very preterm birth <32wks Birth weight (g) Low birth weight (<2500g)

CCS (n¼139)

Non-CCS (n¼308)

P value

37.4  3.9 52 (37.7) 24.2  4.5 7.9  2.6 1.4  0.49 84.2  23.0 39.0  1.7 13 (9.4) 0 (0.0)

35.5  4.0 122 (40.2) 23.7  4.4 7.5  2.6 1.7  0.58 74.2  27.6 39.1  2.1 30 (9.7) 5 (1.6)

<0.001 0.54 0.24 0.09 <0.001 <0.001 0.15 0.91 0.13

3334  549 9 (6.5)

3381  596 19 (6.2)

0.24 0.90

FERTILITY & STERILITYÒ

CONCLUSIONS: Our study is the first to investigate birth outcomes of singleton deliveries with CCS compared to non-CCS FET. Analysis of CCS deliveries revealed no additional risk of trophectoderm biopsy to either fetal growth or birth outcomes compared to non-CCS FETs. In conclusion, babies born after CCS present with excellent perinatal growth and health outcomes. O-33 Monday, October 19, 2015 11:45 AM SELECTION OF SINGLE EUPLOID BLASTOCYSTS FOR TRANSFER VIA PREIMPLANTATION GENETIC SCREENING WITH EITHER NEXT-GENERATION SEQUENCING OR ARRAY COMPARATIVE GENOMIC HYBRIDIZATION: A RANDOMIZED PILOT STUDY. Z. Yang,a J. Lin,b J. Zhang,c Y. Kuang,d J. Liu.e a Research & Development, ZytoGen, Timonium, MD; bART, Reproductive Fertility Center, Irvine, CA; cIVF, New Hope Fertility Center, New York, NY; dART, Shanghai Ninth Hospital, Shanghai, China; eIVF, Jiaen Hospital, Beijing, China. OBJECTIVE: Recent advances in next-generation sequencing (NGS) have provided new methods for preimplantation genetic screening (PGS) of human embryos from IVF cycles. The present study aimed to investigate the effects of NGS screening on clinical pregnancy and ongoing pregnancy outcomes of single embryo transfer for PGS patients in comparison to array comparative genomic hybridization (aCGH) screening. DESIGN: A total of 131 PGS patients at mean age 35.6  3.4 years (ranging 28 to 39 years old) who met the inclusion criteria were randomized into two groups: 1) NGS (Group A): patients (n¼66) had embryos screened with NGS and 2) aCGH (Group B): patients (n¼65) had embryos screened with aCGH. The cohort patients requested PGS due to recurrent pregnancy loss (n¼59), repeated implantation failure (n¼48) and previous aneuploid conceptions (n¼24). MATERIALS AND METHODS: For both NGS and aCGH groups, embryos were cultured to blastocyst stage and trophectoderm biopsy was performed on Day 5 (up to 11:00 pm). Blastocysts were vitrified individually after biopsy. One euploid blastocyst was transferred to individual patients based on the results of PGS with either NGS (Group A) or aCGH (Group B). Clinical pregnancy and ongoing pregnancy rates per transfer were compared between the two groups. Clinical pregnancy was defined as an intrauterine gestational sac with fetal heartbeat visualized by ultrasound examination at week 8 after embryo transfer. Ongoing pregnancy was defined as continuing pregnancy at R 20 weeks of gestation. The categorical variables were analyzed by Chi-square analysis or Fisher’s exact test as appropriate. A two-tailed value of p <0.05 was considered statistically significant. RESULTS: There were no significant differences in female patient’s mean age, Day 3 FSH, AMH, E2, antral follicle number between the two groups (p >0.05). NGS screening identified single euploid blastocysts for transfer and resulted in similar clinical pregnancy rates for PGS patients compared to aCGH screening (69.8% vs. 65.6%, respectively, p >0.05). There was no significant difference in ongoing pregnancy rates between the NGS and aCGH groups (68.3% vs. 63.9%, respectively, p >0.05). The miscarriage rates were also comparable between Group A and Group B (1.5% vs. 1.6%, respectively, p >0.05). CONCLUSIONS: While NGS screening has been recently introduced to assist IVF patients, this is the first randomized clinical study on the efficiency of NGS for single embryo transfer in comparison to aCGH. With the observed high accuracy of 24-chromosome screening and the resulting high clinical pregnancy and ongoing pregnancy rates, NGS has demonstrated an efficient, robust high-throughput technology for selection of single embryo for transfer.

O-34 Monday, October 19, 2015 12:00 PM BLASTOCYSTS NEEDED TO TRANSFER AT LEAST ONE EUPLOID EMBRYO: DATA FROM 10,852 PRE-IMPLANTATION GENETIC SCREENING (PGS) CYCLES. S. Munne,a L. Ribustello,a B. Kolb,b G. Haddad,c J. A. Grifo,d B. Acacio,e Z. Nagy,f J. Zhang,g J. Hesla,h R. J. Kiltz.i aReprogenetics, Livingston, NJ; bMedical DirectorHRC Fertility, Pasadena, CA; cHouston Fertility Institute, Tomball, TX; d NYU Langone Medical Center, New York, NY; eAcacio Fertility Centers, Laguna Niguel, CA; fReproductive Biology Associates, Sandy Springs, GA; gNew Hope Fertility Center, New York, NY; hOregon Reproductive Medicine, Portland, OR; iCNY Fertility Center, Syracuse, NY.

e13