Search

Objective: To report our experience of implementing non-invasive prenatal testing (NIPT) in a tertiary urban academic medical center in South Korea. Materials and methods: An observational retrospective study of singleton and twin pregnancies that underwent prenatal screening for fetal aneuploidy from July 2016 to April 2018 was conducted. Demographics of the study population electing NIPT versus those opting the integrated test were compared. We also assessed clinical significant factors influencing cfDNA fetal fraction in NIPT. Results: Among the 817 women who underwent serum screening tests during the study period, 490 women (60.0%) chose the integrated test while 327 women (40.0%) chose NIPT. Compared to the integrated test group, women in the NIPT group were older (mean age 34.7 ± 3.7 vs. 32.6 ± 3.4; p-value < 0.01), multiparous (47.1% vs. 39.8%; p-value = 0.046), and had higher rate of previous abortion history (28.4% vs. 21.6%; p-value = 0.033). A significant decrease in the number of invasive diagnostic tests was observed since the adoption of NIPT. The screen negative and positive rates of the integrated test group for fetal aneuploidy were 95.3% and 4.7%, respectively while those of the NIPT group were 95.9% and 1.2%, respectively. The rate of inadequate cfDNA fetal fraction was 3.0%. Low fetal fraction was associated with higher maternal age, body weight and BMI. Conclusions: The implementation of NIPT has significantly affected the practice pattern of prenatal aneuploidy screening by replacing the integrated test and decreasing invasive diagnostic tests. Keywords: Non-invasive prenatal testing (NIPT), Integrated test, Maternal serum screening, Tertiary medical center, South Korea

Introduction Trisomy 21 (T21) is the most common aneuploidy affecting humans and is caused by an extra copy of all or part of chromosome 21 (chr21). DNA methylation is an epigenetic event that plays an important role in human diseases via regulation of gene expression. However, the integrative association between DNA methylation and gene expression in T21 fetal placenta has yet to be determined. Methods We profiled expression of 207 genes on chr21 and their DNA methylation patterns in placenta samples from normal and DS fetuses using microarray analysis and predicted the functions of differentially expressed genes using bioinformatics tools. Results We found 47 genes with significantly increased expression in the T21 placenta compared to the normal placenta. Hypomethylation of the 47 genes was observed in the T21 placenta. Most of hypomethylated DNA positions were intragenic regions, i.e. regions inside a gene. Moreover, gene expression and hypomethylated DNA position showed significantly positive associations. By analyzing the properties of the gene-disease network, we found that increased genes in the T21 placenta were significantly associated with T21 and T21 complications such as mental retardation, neurobehavioral manifestations, and congenital abnormalities. Discussion To our knowledge, this is the first study to comprehensively survey the association between gene expression and DNA methylation in chr21 of the T21 fetal placenta. Our findings provide a broad overview of the relationships between gene expression and DNA methylation in the placentas of fetuses with T21 and could contribute to future research efforts concerning genes involvement in disease pathogenesis.

Until now, fetal placenta-specific epigenetic markers for noninvasive prenatal testing of fetal trisomy 21 (T21) have been identified based only on differences in tissue-specific epigenetic characteristics between placenta and maternal blood, but these characteristics have not been validated in T21 placenta. We aimed to discover novel epigenetic markers on chromosome 21 that show a hypermethylated pattern in fetal placenta compared with blood, regardless of the presence of T21. We performed a high-resolution tiling array analysis of chromosome 21 using the methylated-CpG binding domain protein-based method. We identified 93 epigenetic regions that showed fetal placenta-specific differential methylation patterns; among these, three regions showed fetal placenta-specific methylation patterns in T21 placenta samples. The methylation patterns of these three regions in the array were confirmed by bisulfite direct sequencing. The three regions were detectable in first-trimester maternal plasma. Moreover, a combination of their methylation ratio achieved high diagnostic accuracy for noninvasive prenatal testing of fetal T21 by further statistical analysis. These three novel regions with fetal placenta-specific differential methylation patterns on chromosome 21 were identified irrespective of the presence of T21. Our findings suggest that epigenetic characteristics of markers according to the presence or absence of T21 should be considered in the development of noninvasive prenatal testing of fetal T21 using fetal placenta-specific epigenetic markers.

Introduction Down syndrome (DS) is the most common aneuploidy, caused by an extra copy of all or part of chromosome 21 (chr21). Differential microRNA (miRNA) expression is involved in many human diseases including DS. However, the genome-wide changes in miRNA expression in DS fetal placentas have yet to be determined, and the function of these changes is also unclear. Methods We profiled genome-wide miRNA expression in placenta samples from euploid or DS fetuses by using microarray technology and predicted the functions of differentially expressed miRNAs using bioinformatics tools. Results Thirty-four miRNAs were significantly differentially expressed in the DS placenta compared with the normal placenta (16 up-regulated and 18 down-regulated). However, expression of chr21-derived miRNAs did not change. Predicted target genes included 7434 genes targeted by up-regulated miRNAs and 6071 genes targeted by down-regulated miRNAs. Seventy-six of these target genes were located on chr21 (10 genes controlled by down-regulated miRNAs and 34 genes by up-regulated miRNAs, and 32 genes by both). Target genes on chr21 were significantly associated with DS and DS-related disorders, such as mental retardation, neurobehavioral manifestations, and congenital abnormalities. Discussion To our knowledge, this is the first genome-wide study to comprehensively survey placental miRNAs in DS fetuses. Our results provide new insight into miRNA expression in placentas of fetuses with DS. Additionally, our findings indicate that the differentially expressed miRNAs in the DS placenta may potentially affect various pathways related to DS pathogenesis.

To assess whether fetal-derived hypermethylated RASSF1A concentrations in maternal plasma during pregnancy are altered in pregnancies associated with placental dysfunction manifested by intrauterine growth restriction (IUGR), preeclampsia (PE), or placental previa (PP) and whether this alteration can be detected in susceptible subjects before the onset of clinical disease.We performed a real-time quantitative polymerase chain reaction to quantify RASSF1A concentrations before and after methylation-sensitive restriction digestion in maternal plasma at 7-41 gestational weeks of normal pregnancies (n = 161), IUGR (n = 43), PE (n = 22), PP (n = 14) and non-pregnant women (n = 20).A positive correlation was observed between fetal-derived hypermethylated RASSF1A concentration and gestational age for all study groups (r = 0.624, p 0.001 for IUGR; r = 0.381, p = 0.042 for PE; r = 0.697, p 0.001 for PP; r = 0.560, p 0.001 for controls). The concentration of hypermethylated RASSF1A was relatively high at 7-14 gestational weeks in all patient groups. Hypermethylated RASSF1A concentration at 15-28 weeks was significantly higher in patients who subsequently developed IUGR (p = 0.002), PE (p 0.001) or PP (p 0.001) than in controls.We first demonstrated increased concentration of fetal-derived hypermethylated RASSF1A sequences according to advancing gestation and before the onset of the clinical manifestation of pregnancy complications secondary to placental dysfunction, such as IUGR, PE and PP. Hypermethylated RASSF1A in maternal plasma may be useful as a potential biomarker to detect placental-mediated pregnancy complications, regardless of fetal gender and polymorphism.

Objectives: To investigate the risks of increasing maternal age on the perinatal and obstetric outcomes. Materials and Methods: Information about 29,760 singleton pregnancies delivered between 2005 and 2008 was extracted from our database. Patients were categorized into four groups according to age: 20–29 years, 30–34 years, 35–39 years, and ≥40 years. Multivariable logistic regression analysis was used to evaluate the adjusted odd ratios (AORs) of adverse pregnancy outcomes according to maternal age after adjusting for parity, body mass index, medical history and use of in vitro fertilization. Results: The majority of adverse perinatal outcomes were associated with a maternal age ≥35 years as follows: low birth weight (AOR 1.2 and 1.6 for women aged 35–39 years and ≥40 years, respectively); Apgar score

Objective: To determine the incidence of nondisjunction for chromosomes X, Y, and 18 using fluorescence in situ hybridization (FISH) on morphologically normal sperm from infertile men who are candidates for ICSI. Design: After standard hematoxylin staining, sperm with normal morphology were identified using Kruger’s strict morphology criteria. The location of each normal-appearing sperm was recorded using an electronic microstage locator. Slides were subsequently subjected to FISH for detection of chromosomes X, Y, and 18 (control probe). Nuclei were relocated and analyzed under the fluorescent microscope. Setting: University-affiliated IVF and intracytoplasmic sperm injection program. Patient(s): Men classified as infertile on the basis of abnormal strict morphology ( Intervention(s): Semen smears were obtained retrospectively from infertile (n=8) and fertile (n=6) men. Main Outcome Measure(s): Ploidy of each cell was determined according to the number of signals detected for each probe. Result(s): Approximately 100–150 morphologically normal sperm were identified and located in each case. Subsequent FISH analysis of these normal sperm showed aneuploidy to range from 1.8% to 5.5% in the infertile group as compared with 0 to 2.6% among the control fertile group. Statistically significant differences in the incidence of aneuploidy for the sex chromosomes as well as for all three (X, Y, and 18) chromosomes was observed. Conclusion(s): Although 95% to 98% of the sperm were found to be normal for X, Y, and 18, our findings show that infertile couples undergoing ICSI are likely to be at an increased risk for having a genetically abnormal conceptus as compared with the fertile controls. These results demonstrate that normal morphology is not an absolute indicator for the selection of genetically normal sperm. Hence, observed pregnancy failures among ICSI patients may in part be due to the selection of aneuploid sperm.

OBJECTIVE: 1) To examine trends in black-white disparities in infant deaths due to congenital anomalies in the United States, and 2) To explore whether state funding of pregnancy termination for women on Medicaid is associated with rates of infant death due to anomalies. STUDY DESIGN: We used US vital statistics records from 1983-2004. Data on state funding of pregnancy termination under Medicaid was obtained from the Guttmacher Institute. Poisson regression was used to examine rates of infant death due to congenital anomalies by race and year, adjusting for maternal age. Trends in racial disparities were examined by including an interaction term between year and race, then calculating adjusted risk differences (RD) and rate ratios (IRR) at various years. The association between state Medicaid coverage of pregnancy termination for fetal indications and mortality rates in recent years (2001-2004) was examined using a Poisson model. RESULTS: Infant deaths due to anomalies decreased by 3% per year between 1983 and 2004, from 257 to 136 per 100,000 live births. During this time, the gap between blacks and whites widened steadily (Figure). In 1983, the adjusted RD was 20.5 per 100,000 (95% CI [13.8, 27.2], IRR1⁄41.08 [1.06, 1.11]); by 2004, this had grown to 36.2 per 100,000 ([31.5, 40.9], IRR1⁄41.27 [1.24, 1.31]). Black women were more likely to live in states without Medicaid coverage of pregnancy termination (68 vs 59%) and these states had higher rates of infant death due to anomalies (22.0/100,000 higher [15.4, 28.5], IRR1⁄41.19 [1.16, 1.23]). CONCLUSION: Infant deaths due to anomalies are more common among blacks than whites, and this disparity has increased steadily over time. Rates are higher when pregnancy termination for fetal indications is not covered under Medicaid. There is a need to better understand whether differences in access to, or uptake of, pregnancy termination of anomalous fetuses has contributed to the growing black-white gap in infant deaths due to anomalies.

SUBSEQUENTLY SEVERE PREECLAMPSIA SHIN-YOUNG KIM, HYUN-MEE RYU, JAEHYUG YANG, MOON-YOUNG KIM, HYUN-KYONG AHN, JOONG-SIK SHIN, JUNSEEK CHOI, SO-YEON PARK, JIN-MI KIM, BOM-YI LEE, DO-JIN KIM, Samsung Cheil Hospital & Women’s Healthcare Center, Laboratory of Medical Genetics, Seoul, Korea, South Korea, Samsung Cheil Hospital & Women’s Healthcare Center, Sungkyunkwan University School of Medicine, Obstetrics and Gynecology, Seoul, Korea, South Korea OBJECTIVE: It has been shown that maternal serum inhibin-A levels are increased in women with preeclampsia. Little is known about inhibin-A levels in amniotic fluid. The aim of this study was to determine whether amniotic fluid and maternal serum inhibin-A levels are elevated in the early second trimester of women who subsequently develop severe preeclampsia. STUDY DESIGN: We studied the levels of inhibin-A in amniotic fluid and maternal serum at 15-20 weeks’ gestation from normal pregnant women (n = 80) and from pregnant women with subsequently severe preeclampsia (n = 50). All samples were assayed for inhibin-A by enzyme-linked immunosorbent assay (ELISA). RESULTS: Amniotic fluid and maternal serum inhibin-A levels were significantly increased in severe preeclampsia compared to normal pregnant women (P ! .001 for both). With a specific cutoff value, the estimated odds for severe SMFM Abstracts S35

SCREENING CHROMOSOMAL ABNORMALITY IN KOREAN POPULATION JAE HYUG YANG, MIN HYOUNG KIM, JIN HOON CHUNG, HYUN KYONG AHN, MOON YOUNG KIM, HYUN MEE RYU, JUNE SEEK CHOI, Samsung Cheil Hospital and Women’s Healthcare Center, Sungkyunkwan University, School of Medicine, Obstetrics and Gynecology, Seoul, South Korea, Samsung Cheil Hospital and Women’s Healthcare Center, Sungkyunkwan University, School of Medicine, Obstetric and Gynecology, Seoul, South Korea OBJECTIVE: This study was aimed to compare the sensitivities and false positive rates of screening test for chromosomal abnormality using ultrasonographic measurement of nuchal translucency(NT) with different cut-offs in Korean population. STUDY DESIGN: Between January 2001 and December 2001, 2688 singleton pregnancies undergoing ultrasound between 11 and 14 weeks of gestation were involved in this study. We reviewed the obstetrical and neonatal medical records and analyzed the sensitivities and false positive rates of NT for screening chromosomal abnormalities using three cut-offs such as 2.5 mm, 3.0 mm, and 95th percentile for each crown-rump length (CRL). RESULTS: There were 32 chromosomal abnormalities (1.2%) including 12 cases of trisomy 21. The numbers of chromosomal abnormalities detected by NT with different cut-offs of 2.5 mm, 3.0 mm and 95th percentile were 22, 18 and 23 respectively. By using cut-off as 95th percentile, we could detect additional one more case of trisomy 18 than using fixed cut-off, 2.5 mm. The sensitivities and false positive rates for all chromosomal abnormalities were 68.8%, 56.3%, 71.8% and 6.4%, 3.6%, 5.7% respectively according to each cut-off, 2.5 mm, 3.0 mm and 95th percentile. The sensitivities of trisomy 21 were 75.0%, 50.0%, and 75.0%, respectively. CONCLUSION: The cut-off using 95th percentile for each fetal CRL increases the sensitivity and decreases false positive rate to detect the chromosomal abnormality compared with a fixed cut-off. CRL dependent cut-off of NT may be the more rational one than a fixed cut-off for screening chromosomal abnormality in Korean population. 644 MULTIFETAL PREGNANCY REDUCTION EFFECT OF NUCHAL TRANSLUCENCY ASSESSMENT: THE THOMAS JEFFERSON UNIVERSITY EXPERIENCE, 2001-2004 SHARON BYUN, SURASITH CHAITHONGWONGWATTHANA, WARALAK YAMASMIT, KEITH RYCHLAK, STUART WEINER, Thomas Jefferson University, Department of Obstetrics and Gynecology, PHILADELPHIA, Pennsylvania OBJECTIVE: To report our experience of pregnancy outcome after multifetal reduction (MFR) with the routine use of nuchal translucency (NT) measurements. STUDY DESIGN: We performed a retrospective review of all patients who underwent MFR at Thomas Jefferson University Hospital between 1/1/01-2/13/ 04 (N = 142). Reduction selection criteria were by fetal position, CVS, abnormal NT, or other indications. Pregnancy outcome data were obtained for preterm labor, preterm premature rupture of membranes, gestational hypertension, and gestational diabetes; gestational age at delivery, birth weight, mode of delivery, maternal postpartum complications and neonatal complications. RESULTS: 142 patients underwent MFR during this time period. 16/391 fetuses had abnormal NT measurements (4.09%). Of the 375 fetuses that had normal NT measurements, 23.9% underwent CVS. Of the 16 fetuses that had increased NTs, 20.0% had CVS (P = NS). CVS was performed on 116 fetuses. 8/102 CVS results that were available had an abnormal karyotype (7.8%). Reduction selection was by fetal position in 47.4%, by CVS in 25.2%, by increased NT in 4.4%, due to anomalies in 4.4%, by selecting the monochorionic pair in 7.4%, and by some combination in 11.0%. Pregnancy outcome data were obtained for the delivered 94 pregnancies. There was one first trimester loss (1.1%). The overall mean gestational age at delivery was 34.9 weeks. The mean birth weight was 2192 g (SD 711 g). CONCLUSION: There was a low rate of pregnancy loss following multifetal reduction. There was not a statistically significant difference in maternal complications rates, mean gestational age at delivery, and neonatal birth weight when reduction was by NT, CVS, and position. There was not a statistically significant decrease in the number of CVSs performed when there was an increased NT measurement. However, further investigation is needed to determine whether NT measurements alone could be used to aid in selective multifetal reduction by directing CVS toward the more likely euploid embryo and/or directing MFR toward the more likely abnormal one.

(MTHFR C677T AND A1298C) AND METHIONINE SYNTHASE REDUCTASE (MTRR A66G) GENE AS MATERNAL RISK FACTORS FOR FETAL ANEUPLOIDY HYUNMEE RYU, DO-JIN KIM, MOON-YOUNG KIM, SO-YEON PARK, JIN-WOO KIM, SHIN-YOUNG KIM, JAE-HYUG YANG, JOUNG-YEOL HAN, JOO OH KIM, JINHOON CHUNG, Samsung Cheil Hospital and Women’s Healthcare Center, Sungkyunkwan University School of Medicien, Obstetrics and Gynecology, Seoul, Korea, South Korea, Samsung Cheil Hospital and Women’s Healthcare Center, Laboratory of Medical Genetics, Seoul, Korea, South Korea OBJECTIVE: The purpose of this study was to address the association of MTHFR (C677T/A1298C) and MTRR (A66G) polymorphisms as maternal risk factors for fetal aneuploidy. STUDY DESIGN: We studied the presence of MTHFR C677T/A1298C and MTRR A66G in an aneuploidy group (n = 30) who had fetal aneuploidy and in a control group (n = 78) who had given birth to at least two healthy children without a history of miscarriage or abnormal pregnancy. To determine the polymorphism of these enzymes, we performed PCR/RFLP (Restriction Fragment Length Polymorphism). RESULTS: There were no statistical differences in the prevalence of MTHFR 677 variant T allele (P = .519) and 1298 variant C allele (P = .051) in both groups. Frequency of MTRR G allele in aneuploidy group was higher than in control group. Mother with MTRR 66 AG mutation had a 2.9-fold higher risk of having a fetus with aneuploidy than mother without the G substitute (odds ratio: 2.9; 95% CI: 1.5-5.3; P = .001). CONCLUSION: This study suggests that there was an increased risk of fetal aneuploidy in mother carriers of MTRR G allele, not in MTHFR C677T and A1298C. The MTRR A66G variant may be a significant risk factor for producing a child with chromosome aneuploidy.