should embryos screened for single gene disorders undergo concurrent aneuploidy screening?

GENETIC COUNSELING

O-155 Monday, October 14, 2013 04:00 PM

FEMALE CYSTIC FIBROSIS CARRIERS AND ASSISTED REPRO-DUCTIVE TECHNOLOGIES - IS THERE A CORRELATIONBETWEEN MUTATION LOCI AND RESPONSE TO IN VITROFERTILIZATION STIMULATION? T. A. VanWort,a A. Esmaeili,b

J. A. Lee,a M.Whitehouse,a N. Bar-Chama,a,b A. B. Copperman.a,b aRepro-ductive Medicine Associates of New York, New York, NY; bObstetrics,Gynecology and Reproductive Science, Mount Sinai School of Medicine,New York, NY.

OBJECTIVE: Cystic Fibrosis (CF) is an autosomal recessive genetic dis-ease that can affect reproductive organs. Mutations of the cystic fibrosistransmembrane conductance regulator gene in male carriers can be associ-ated with Congenital Bilateral Absence of the Vas Deferens, yet there areno known adverse effects on the fertility of female carriers. To elucidateany association, we compared female carriers by mutation loci and assessedtheir response to in vitro fertilization (IVF).

DESIGN: Retrospective study.MATERIALS AND METHODS: IVF cycles of female CF carriers

(n¼280; 2002-2013) were included. Cohorts were classified by mutationloci (delta F508, W1282X, R117H, G542X) and compared by the variablesbelow. Chi-square and Kruskal–Wallis analysis of variance ranks were per-formed with significance at p<0.05.

RESULTS: Significant differences in age, follicle stimulating hormone,estradiol levels and day of embryo transfer were observed yet locus - specificpregnancy outcomes did not significantly differ.

CONCLUSION: Our study showed significant differences in IVF responseamong female CF carrier loci. Despite comparable pregnancy rates, weconsider loci response variance to be relevant. Possible diagnostic and treat-ment algorithms could personalize our approach for carriers based on muta-tion loci. Further investigation will expand our understanding of thebiological mechanisms controlled by each locus and how they affect cyclestimulation response and outcome.


AN ANALYSIS OF COMPREHENSIVE CARRIER SCREENING RE-SULTS FOR 1000 CLINICAL SAMPLES: THE IMPORTANCE OFTRANSPARENT DISEASE CLASSIFICATION & CRITERIA FORSELECTION. A. Bisignano,a N. Kumar,a R. N. Prates,b S. Munne,b

J. Grifo,c D. Hoffman.d aRecombine LLC, New York, NY; bRecombineLLC and Reprogenetics LLC, Livingston, NJ; cNew York University Schoolof Medicine, New York, NY; dIVF Florida Reproductive Associates,Margate, FL.

OBJECTIVE: Advancements in genomics allow for expanded, cost-effec-tive, and high-throughput carrier screening. The American College of Med-ical Genetics (ACMG) recommends that phenotype and penetrance should beconsidered in selecting diseases for inclusion. Based on 1000 clinical sam-ples screened, the selected diseases were classified according to these factorswith the goal of improving transparency, pre- and post-test counseling andresults disclosure protocols.

DESIGN: Retrospective study.

delta F508 (n¼192) W1282X (n¼37)

Age 39.2 � 13.1 35.6 � 5.4FSH 10 � 6** 8 � 4.7**Peak E2 (pg/mL) 1801.5 � 1097.4** 2420 � 1463.6**# Oocytes 11.5 � 9.2 14.7 � 9.4#MII 5.4 � 7.6 8.7 � 7.3Fert Rate 69.5% 64.4%D3 43.5% 17.7%D5 ET 47.8% 73.5%Clinical Preg Rate 45% 40.5%Delivery Rate 25.5% 29.7%

(** ¼ significance between loci).

FERTILITY & STERILITY�

MATERIALS AND METHODS: Recombine’s Comprehensive CarrierScreen for 978 mutations associated with 181 recessive diseases was per-formed on 1000 clinical referrals. Diseases were classified into 2 groups:high impact (significant effect on quality of life/reduced lifespan) and vari-able spectrum (less severe phenotype/lower penetrance). Informed consentto utilize de-identified data was obtained from all patients.RESULTS: See table

Most Common Variable Spectrum

R117H (n¼27) G542X (n¼14)

38.7 � 3.2** 33.5 � 3.5**12.4 � 3.1** 8.7 � 1.6**

1289.7 � 660.8** 2147.1 � 96010.8 � 4 12.3 � 8.75.2 � 5.1 7.6 � 8.370.8% 62.6%69.6% 42.9%17.4% 57.14%44.4% 64.3%33.3% 57.1%

%

p-v

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1 in
TOTAL 98.9 1 MTHFR Deficiency 53.3 2 Hemochromatosis: Type 1 24.1 4 AMPD Deficiency 15.3 7 Duarte Galactosemia 13.8 7 Factor V Deficiency 9.9 10 Familial Mediterranean Fever: Mild 9.8 10 Congenital Nonclassical Adrenal Hyperplasia 9.8 10 Most Common High-Impact % 1 in TOTAL 39.9 3 Familial Mediterranean Fever 5.9 17 Biotinidase Deficiency 5.3 19 Nonsyndromic Hearing Loss and Deafness 4.9 20 Polycystic Kidney Disease 3.4 29 Cystic Fibrosis 3.2 31 Glycogen Storage Disease: Type II 1.2 81 Alpha Thalassemia 1.2 81
CONCLUSION:With the full panel,�99% of patients carry at least 1 mu-tation. More than 50% of samples carry MTHFR mutations. Consideringonly high-impact diseases,�40% of patients carry at least 1 mutation. Inclu-sion of mild disorders may create anxiety and logistical concerns for patientsand physicians. This anxiety can be reduced with post-test genetic coun-seling, transparency about differences in severity between high-impact andvariable spectrum diseases in all materials (pre-counseling, web-based &reports), and the ability to customize the disease panel.


SHOULD EMBRYOS SCREENED FOR SINGLEGENEDISORDERSUNDERGO CONCURRENT ANEUPLOIDY SCREENING? K. N.Goldman, A. Adler, H.-L. Lee, E. Ampeloquio, J. A. Grifo. Obstetricsand Gynecology, Reproductive Endocrinology and Infertility, New YorkUniversity Langone Medical Center, New York, NY.

OBJECTIVE: To determine the incidence of aneuploidy in a population ofpatients undergoing pre-implantation genetic diagnosis (PGD) for singlegene disorders (SGD).DESIGN: Retrospective analysis.MATERIALS ANDMETHODS: We analyzed in vitro fertilization cycles

from 2011-2012 in which patients underwent their 1st cycle of PGDwith tro-phectoderm (TE) biopsy for SGD with concomitant array-comparativegenomic hybridization (aCGH) for aneuploidy screening. Data are presentedin mean � SD.

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0.050.050.050.30.050.40.050.050.70.05

S47

RESULTS: 19 patients underwent PGD/TE biopsy for SGD + aCGHfor the following indications: BRCA (n¼4), Fragile X (n¼4), Gaucher Dis-ease (n¼3), Dystonia, Hereditary Angioedema, Arthrogryposis, IncontinentiPigmentosa, Holt-Oram Syndrome, Familial Dysautonomia, Beta-Thalas-semia, and HLA matching. Mean age was 32.1 � 7.4 y, and mean folliclestimulating hormone and estradiol on day 2 were 6 � 3 IU/L and 41 � 16pg/ml, respectively. The mean number of oocytes and MII oocytes was 19� 10 and 16 � 9, respectively, and the number of 2PN embryos was 13 �7.4. The blastocyst formation rate was 60% (149/248). 149 blastocysts(day-5¼108; day-6¼41) underwent TE biopsy, with 7.8 embryos biopsiedper patient. While 15 patients had embryos (n¼34) that were both unaffectedby a SGD and euploid, the majority of patients (68%) had R 1 embryo thatwas unaffected by a SGD but aneuploid. 12 patients (63%) underwent frozenembryo transfer (FET), with the majority having a single ET (75%). Implan-tation rate was 80% (12/15), spontaneous abortion rate was 9% (1/11 clinicalpregnancies), and live birth/ongoing pregnancy rate was 75% (9/12). 4 pa-tients (21%) had no normal (unaffected/euploid) embryos available for ET.3 patients (16%) have not yet returned for ET.

CONCLUSION: The majority (68%) of patients undergoing SGD testingplus aneuploidy screening had R 1 embryo(s) that were unaffected by theSGD but also aneuploid. Patients and providers should understand the riskof transferring aneuploid embryos when SGD testing is performed withoutaneuploidy screening.


LOWAMHLEVELSDONOT PREDICTAHIGHER INCIDENCEOFANEUPLOID BLASTOCYSTS IN YOUNG INFERTILITY PATIENTS(<35 YEARS). S. McCormick, M. Linden, D. Young, D. Klepacka,W. B. Schoolcraft, M. G. Katz-Jaffe. Colorado Center for ReproductiveMedicine, Lone Tree, CO.

OBJECTIVE: Antimullerian hormone (AMH) testing is utilized clinicallyto predict ovarian response to gonadotrophin stimulation and IVF outcome.This study investigated the relationship in young infertility patients (<35years) with relatively low AMH levels (%1.4ng/ml) and blastocyst chromo-some numeration.

DESIGN: IRB approved research study.MATERIALS AND METHODS: Young infertility patients (<35 years;

n¼206) with either normal AMH levels (>1.4ng/ml; n¼154) or relatively lowAMH levels (%1.4ng/ml; n¼52) underwent routine IVF treatment. At the blas-tocyst stage, a biopsy was performed for chromosome numeration using eitherquantitative PCR or SNP microarray (RMA-NJ), prior to embryo vitrification.Euploid blastocysts were warmed and transferred in a frozen embryo transfer.

RESULTS: Overall the proportion of aneuploid blastocysts for younginfertility patients was independent of AMH levels (>1.4¼29.7% vs.%1.4¼29.4%; ns). There were also no significant differences between theAMH groups for maternal age, D3 FSH, blastocyst survival post vitrification,blastocyst gender or the number of chromosome errors observed per blasto-cyst. However, low AMH levels did reflect a significantly reduced number ofoocytes retrieved (>1.4¼23.5 �8.8 vs. %1.4¼16.5 �5.3; P<0.0001) andblastocysts biopsied (>1.4¼8.1 �4.5 vs. %1.4¼5.1 �2.2; P<0.0001).Following transfer of euploid blastocysts there were no differences in out-comes between the AMH groups for implantation rate with fetal cardiac ac-tivity (>1.4¼72.1% vs.%1.4¼75.6%; ns) or live birth rate (>1.4¼77.8% vs.%1.4¼79.6%; ns). As only 8% of young infertility patients had abnormal D3FSH, it was not considered as an independent variable.

CONCLUSION: A low AMH level in younger infertility patients is asso-ciated with a poorer ovarian response but is not predictive of a higher inci-dence of chromosomally aneuploid blastocysts. Indeed, chromosomallyeuploid blastocysts identified for transfer resulted in equivalent implantationpotential and live birth independent of maternal AMH levels.


INTRAUTERINE HIGH-GLUCOSE ENVIRONMENT INFLUENCESPLACENTAL DEVELOPMENT THROUGH IMPRINTING REGU-LATION OF DLK1/GTL2 REGION. Y. Jiang,a Q. Luo,a Q. Gao,b

G. Ding,a J. Sheng,b H. Huang.a aReproductive Endocrinology Department,Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou,Zhejiang, China; bPathophysiological Department, School of Medicine, Zhe-jiang University, Hangzhou, Zhejiang, China.

OBJECTIVE: Gestational Diabetes Mellitus offspring has a higher ten-dency of chronic adult disease.We supposed these diseases may be linked

S48 ASRM Abstracts

to abnormal placental development and function, and sought to identifythe molecular and morphological changes in GDM mice.DESIGN: Observed placental development in F1-GDM mice and in

F2-offspring(by intercrossed between control mice and F1-GDM mice).Moreover, we detected the expression levels of imprinted gene Dlk1/Gtl2on mouse chromosome 12. Furthermore, we examined methylation statusof Dlk1-DMR, IG-DMR and Gtl2-DMR.MATERIALS AND METHODS: Mouse model of GDM was induced by

STZ on the day of pregnancy. Mouse placentae were collected on embryonicday 12.5,15.5, 18.5, and histology analysis was done by HE staining. Dlk1-DMR, IG-DMR and Gtl2-DMR methylation was detected by BisulfiteSequencing. Gene expression was assayed by quantitative PCR.RESULTS: 1) Both placenta weight in F1-GDM group were significantly

lower than control group on any embryonic day. However, there were no dif-ference in F2-offspring placenta. Neither did Histology analysis. 2)Compared with control group, in both F1-GDM and F2-GDM placentae,Dlk1 RNA expression level was lower (P<0.05), while Gtl2 RNA expressionlevel was higher (P<0.05); Both F1-GDM and F2-GDM placentae, Dlk1-DMR was relatively high methylation state, however Gtl2-DMR and IG-DMR showed lower methylation state. 3) Compared with normal glucoseconcentration group, we gained same tendency for Dlk1 and Gtl2 expressionin vitro.CONCLUSION: Intrauterine high-glucose environment of GDM leaded to

morphology alteration in the first generation,but not in second generation.However, in the molecular aspects, both the first and second generationwere affected, and there was parental imprinting in the second generationfor both Dlk1 and Gtl2 expression. Meanwhile,the methylation state ofDlk1-DMR/ IG-DMR/ Glt2-DMR in both F1-GDM and F2-GDM groupmay be linked with gene expression.Supported by: National Scientific Foundation of China (No.30973209).


A NOVEL ROLE FOR GENOMIC COPY NUMBER VARIANTS INFEMALE INFERTILITY. T. Hu-Seliger,a M. Elashoff,a J.-M. Chia,a

H. J. Kang,b Z. Rosenwaks,b P. Yurttas Beim.a aCelmatix Inc., New York,NY; bRonald O. Perelman and Claudia Cohen Center for Reproductive Med-icine, Weill Cornell Medical College, New York, NY.

OBJECTIVE: Copy number variants (CNVs) are genomic aberrations thatalter the number of copies of genomic segments and therefore have the po-tential to alter dosage-sensitive genes and/or functional elements. Recentstudies have shown that CNVs underlie various complex spectrum disordersand that the severity of the disorder is correlated with overall CNV burden. Toinvestigate the role of CNVs in female infertility spectrum disorders, we haveperformed a survey examining CNVs in a cohort of patients with unexplainedinfertility.DESIGN: Retrospective cohort of femalepatientswith unexplained infertility.MATERIALS AND METHODS: Genomic DNA extracted from blood

samples was obtained from patients undergoing non-donor in-vitro fertiliza-tion (IVF) cycles at a large academic medical center. CNVs were detected us-ing a custom NimbleGen Comparative Genomic Hybridization (CGH)Whole-Genome Tiling Array optimized for fertility-related loci. The resultinglog2 ratios of the raw intensities were then normalized and segmented. Theprevalence of CNVs among the infertile cohort was compared to the Child-ren’s Hospital of Philadelphia (CHOP) CNV database of healthy individuals.RESULTS: Approximately30%of the>3000CNVsdetectedwereunique to

the infertile patients (P<10-7, Binomial Test of Proportions). Moreover, CNVsspecific to the infertile cohort appear to be overrepresented among regions of thegenome predicted to play a role in mammalian fertility (P<10-16, t-test).CONCLUSION: The set of CNVs uniquely observed in our female unex-

plained infertility cohort represent putativegenetic biomarkers thatmaypredicta range of infertility conditions difficult to diagnosewith existing clinical data.Supported by: Celmatix Inc.


RELATIONSHIPBETWEENGP130 (IL6ST)GENEPOLYMORPHISMGENOTYPES AND RECURRENT MISCARRIAGE. A. L. Mauri,a,b

L. D. Vagnini,b C. G. Petersen,a,b,c J. B. A. Oliveira,a,b,c R. L. R. Baruffi,a,b

J. G. Franco, Jr.a,b,c aCenter for Human Reproduction Prof. Franco Jr, RibeiraoPreto, Sao Paulo, Brazil; bPaulista Center for Diagnosis Research and Training,Ribeirao Preto, Sao Paulo, Brazil; cDepartment of Gynecology and Obstetrics,BotucatuMedical School -Sao Paulo State University - UNESP, Botucatu, SaoPaulo, Brazil.

Vol. 100, No. 3, Supplement, September 2013

should embryos screened for single gene disorders undergo concurrent aneuploidy screening?

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