Healthy baby after intrauterine transfer of monosomic embryos

CONCLUSIONS: This report details a collaborative pathway providing couples an opportunity to: i) build a family without continuation of a monogenic familial disease; ii) donate otherwise discarded embryos for derivation of ds-hESCs; and thus iii) establish a universally-shared resource to study molecular causes and consequences of familial monogenic diseases and discover future treatments and/or cures of diseases that affect(ed) their living/deceased relatives. P-145 Tuesday, October 18, 2016 HEALTHY BABY AFTER INTRAUTERINE TRANSFER OF MONOSOMIC EMBRYOS. P. Rubino, L. Dearden, L. Guan, R. Ruiz De Assin, K. Mazmanian, B. A. Kolb, J. Nelson, J. M. Norian, J. Wilcox, T. Tan. HRCFertility, Pasadena, CA. OBJECTIVE: Full autosomal monosomy embryos are lethal, so they don’t implant or result in early pregnancy loss. In cases where no euploid embryos are available, the transfer of monosomic embryos might be considered. The objective of this study was to report the results in which only monosomic embryos were transferred. DESIGN: Retrospective analysis of 13 frozen embryo transfers (FET) between January 2013 to April 2016 in which only full monosomic embryos were transferred. MATERIALS AND METHODS: Multiple trophectoderm cells were biopsied on day 5/6 blastocysts and sent for preimplantation genetic screening (PGS) by array-comparative genomic hybridization (aCGH). Embryos were vitrified and used for subsequent FET. If no, or inadequate number of euploid embryos were present, the patient was offered the transfer of selected monosomic embryo(s) for FET. Patients were counseled of risk and signed an informed consent. RESULTS: From January 2013 to April 2016, 1076 FET-PGS cycles were performed. The transfer of monosomic embryos was made available for 13 women whom IVF had resulted in no euploid embryos. The clinical outcomes are reported in the table I.

Table I. Clinical Outcomes of Monosomic Blastocysts Transferred

Patient n

Karyotype of transferred embryos

Clinical outcome

1 2 3 4 5 6 7 8 9 10 11 12 13

45, XX, -4 45, XY, -22 45, XY, -1 45, XX, -8; 45, XX, -21 45, XY, -5 45, XY, -5 45, XX, -21; 45, XY, -22 45, XX, -5 45, XY, -22; 45, XY, -16 45, XX, -21 45, XX, -4 45, XY, -15 45, XY, -19

Baby healthy at birth No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy No pregnancy

CONCLUSIONS: A concern of PGS is ‘false positive’ embryos being discarded or ‘false negative’ embryos transferred. In our study an embryo that was diagnosed as full monosomy 4 was transferred resulting in a healthy live birth. A possible explanation for the misdiagnosis may be mosaicism, where different genetic cell lines are segregated in the trophectoderm of the embryo, while the inner cell mass contained only euploid cells. Sampling errors during biopsy and the lack of sensitivity necessary to detect minor cell populations makes mosaicism difficult to detect for aCGH, resulting in misdiagnosis. aCGH in PGS is still not fully definitive in the diagnosis of aneuplody. The patient must be approprietely counseled of risk, requirement of prenatal screening and possibility of termination of pregnancy before the transfer of full autosomal monosomic embryos. References: 1. Munne S, Grifo J, Wells D. Mosaicism: ‘‘survival of the fittest’’ versus ‘‘no embryo left behind’’ Fertil Steril. 2016 May;105(5):1146-9. 2. Greco E, Minasi MG, Fiorentino F. Healthy babies after Intrauterine transfer of Mosaic Aneuploid Blastocyst N Engl J Med. 2015 Nov 19;373(21):2089-90.

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ASRM Abstracts

P-146 Tuesday, October 18, 2016 DIAGNOSTIC AND CLINICAL OUTCOMES OF 694 CYCLES USING KARYOMAPPING FOR PREIMPLANTATION GENETIC DIAGNOSIS (PGD) OF SINGLE GENE DISORDERS. D. GoldbergStrassler,a R. Cabey,a A. Jordan,a R. Prates,b E. Mounts,c e. barbieri,c A. Hershlag,d M. Guarnaccia,e M. Surrey,f S. Munne.a aReprogenetics, Livingston, NJ; bMolecular, Reprogenetics, Livingston, FL; cOregon Reproductive Medicine, Portland, OR; dHofstra Northwell School of Medicine, Manhasset, NY; eREI, Columbia University, NY, NY; fSouthern California Reproductive Center, Beverly Hills, CA. OBJECTIVE: Karyomapping (Kmap) is a linkage-based single nucleotide polymorphism technology proven to be highly efficient in PGD diagnosis and applicable to most single gene disorders (SGD). Here, outcomes of such testing are reviewed. DESIGN: Kmap with and without comprehensive chromosome screening (CCS) via array comparative genomic hybridization (aCGH) (24sure, Illumina) or next generation sequencing (NGS) (VeriSeq, Illumina) was used to screen embryos (blastocysts) undergoing PGD for SGDs. MATERIALS AND METHODS: Between 1/2014-4/2016, PGD was performed on 694 cycles (4284 blastocyst biopsies) with subsequent cryopreservation. Each sample was whole genome amplified and analyzed using Kmap (Illumina, USA). Additionally, 96.1% (609/694) of cases had CCS. Followup data was obtained for 478 cycles with embryos suitable for transfer (free from SGD and euploid if CCS performed). At the time of data collection, 223 cycles had undergone embryo transfer. RESULTS: The 694 Kmap cycles comprised 105 SGDs, human leukocyte antigen (HLA) matching, and microdeletions/duplications. Diagnostic results were available for 97% (4154/4284) of samples. Implantation rate was 75.9% (151/199) for PGD+CCS (average maternal age (MA): 32.9 years) and 64% (16/25) for PGD only (average MA: 34.3 years). The pregnancy rate per transfer was 74.7% (124/166) for PGD+CCS and 78.9% (15/19) for PGD only. Single embryo transfer was performed for most; 20 had double embryo transfer. The average number of embryos suitable for transfer per cycle was 1.95 (PGD+CCS) and 2.56 (PGD only). A total of 30 live births and 96 ongoing pregnancies were reported. Confirmatory testing via chorionic villus sampling (CVS) or amniocentesis was performed for 6 pregnancies. Follow-up testing via newborn screening panel for 9 live births was in concordance with Kmap diagnosis. Results were in complete concordance with Kmap diagnosis; no misdiagnoses have been reported to date from the 121 cycles with successful pregnancy outcomes. CONCLUSIONS: With the increase in patient awareness regarding availability of PGD and the rise of preconception carrier screening, there is a growing demand for PGD. Higher implantation rates were observed for patients who underwent CCS in combination with Kmap. Pregnancy rates were similar as sample size for ‘‘PGD only’’ was smaller and 62.5% of ‘‘PGD only’’ patients had a double embryo transfer. Due to the high diagnostic accuracy, comprehensive analysis and short preparation time, Kmap is a successful treatment strategy for patients requesting PGD for inherited disorders. P-147 Tuesday, October 18, 2016 VALIDATION OF DETECTING MONOGENETIC DISEASES & ANEUPLOIDY SIMULTANEOUSLY BY NEXT GENERATION SEQUENCING WITH LINKAGE ANALYSIS. S. Lu,a J. Qiao,b X. Xie.c aYikon Genomics Co., Ltd., Shanghai, China; bDepartment of Obstetrics and Gynecology, Third Hospital, Peking University, Beijing, China; cHarvard University, Cambridge, MA. OBJECTIVE: Here we report three supportive cases by screening embryos with our well-established and published method, named ‘‘mutated allele revealed by sequencing with aneuploidy and linkage analyses’’ (MARSALA), which combines multiple annealing and looping based amplification cycles (MALBACTM) and Next Generation Sequencing with linkage analysis. The MARSALA strategy is able to detect aneuploidy and targeted gene mutations simultaneously in preimplantation genetic diagnosis (PGD) process during an in vitro fertilization (IVF) cycle. DESIGN: We sequence the genome of the parents and relatives carrying the monogenic disease allele with the depth of 2x genome coverage. Then, from the sequencing results of each embryo, the SNP readouts (heterozygous or homozygous) adjacent to the disease-cause mutation sites allowed the identification of the disease-carrying allele in the embryo. MATERIALS AND METHODS: Blastocyst Biopsy We collected a few TE cells from each hatching blastocyst on day 5 or day 6. All embryos

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