Journal of Gynecology Obstetrics and Human Reproduction 48 (2019) 521–525 Available online at ScienceDirect www.sciencedirect.com Original Article The type and prevalence of chromosomal abnormalities in couples with recurrent first trimester abortions: A Turkish retrospective study a, b a a Malik Ejder Yildirim *, Savas Karakus , Hande Kucuk Kurtulgan , Burak Baser , a Ilhan Sezgin a Cumhuriyet University, Faculty of Medicine, Department of Medical Genetics, Turkey b Cumhuriyet University, Faculty of Medicine, Department of Obstetrics and Gynecology, Turkey A R T I C L E I N F O A B S T R A C T Article history: Objective: Chromosomal abnormalities are more common in the first trimester abortions. We aimed to Received 8 March 2019 investigate the types and prevalence of chromosomal abnormalities in couples with recurrent first Received in revised form 2 May 2019 trimester miscarriages in Sivas, Turkey. Accepted 7 May 2019 Materials and medhods: Three hundred couples (600 individuals) who had a story of recurrent abortion Available online 11 May 2019 were included in the study. Chromosome analysis was performed after the preparation of lymphocyte culture with the standard method. Karyotype analyses were supported by FISH and aCGH studies. Keywords: Results: Total 26 chromosome abnormalities (8.7%) were found in the couples (19 females and 7 males). Recurrent abortion Fifteen cases (57.7%) were structural anomalies and eleven cases (42.3%) were numerical chromosomal Chromosomal abnormality Heterochromatin aberrations. We detected 5 balanced translocations (33.3%), 4 Robertsonian translocations (26.7%), 3 Satellite inversions (20%), 2 duplications (13.3%) and one deletion (6.7%) among the structural anomalies. Mosaic monosomy X in five cases (45.4%), the combination of mosaic monosomy-trisomy X in three cases (27.3%), the combination of mosaic monosomy-trisomy and tetrasomy X in two cases (18.2%) and mosaic pentasomy X in only one individual (9.1%) were encountered as numerical chromosome aberrations. 19 cases had heterochromatic changes or other chromosomal variations (satellite increments and inv9). Conclusion: Chromosome analysis in couples with recurrent miscarriage is necessary for possible preimplantation genetic diagnosis. As well as numerical and structural chromosome abnormalities, some chromosomal variations (heterochromatin and satellite increments etc.) may also contribute to recurrent miscarriages. Numerical chromosomal abnormalities are often associated with sex chromosomes and usually seen in females. © 2019 Elsevier Masson SAS. All rights reserved. Introduction who are 35 years old and over [3,4]. If the main cause of pregnancy loss cannot be explained; this abortion may be evaluated as Recurrent miscarriage (RM) may be defined as two or three and idiopathic or unexplained spontaneous abortion. more consecutive pregnancy losses within the first 20 weeks of Chromosome anomalies are important especially in early gestation and include approximately 0.8%–1.4% of pregnancies [1]. abortions. The majority of such abnormalities may be due to There may be many different reasons for this problem, but a chromosomal non-disjunction, translocation or another mutation possible cause can be identified in only about 50% of women with [5]. A balanced or unbalanced karyotype in one of the partners in a recurrent miscarriage [2]. The etiology of recurrent abortions may couple as a structural chromosomal abnormality (reciprocal or include endocrine or immunological disorders, parental chromo- Robertsonian translocations etc.) may result in recurrent miscar- somal anomalies, hypercoagulability and anatomic abnormalities riage, or physical and/or mental disorder in next generation [6]. and these consecutive miscarriages are more common in women Numerical chromosome anomalies are also a major cause of abortion. Some researchers suggested that the most common chromosomal abnormality observed within first trimester sponta- * Corresponding author at: Cumhuriyet University, Faculty of Medicine, neous abortions may be single trisomies in abortion materials [7]. Department of Medical Genetics, 58104, Sivas, Turkey. This problem may be associated with a balanced translocation in E-mail addresses: [email protected] (M.E. Yildirim), parents. The percentage of chromosomal variations was expressed [email protected] (S. Karakus), [email protected] as 5.5% in couples with abortions compared to 0.55% of the general (H.K. Kurtulgan), [email protected] (B. Baser), [email protected] (I. Sezgin). population [8]. http://dx.doi.org/10.1016/j.jogoh.2019.05.014 2468-7847/© 2019 Elsevier Masson SAS. All rights reserved. 522 M.E. Yildirim et al. / J Gynecol Obstet Hum Reprod 48 (2019) 521–525 We analyzed the frequency and type of chromosomal abnor- records were examined, it was found that 48% of the cases were malities in recurrent miscarriage in this study. chemical pregnancy, 33% were spontaneous abortion, 12% were missed abortion and 7% of them were blighted ovum. On average, Materials and methods the gravidity, parity and abortion numbers were 4, 1 and 3, respectively (Table 1). Among 300 couples, 26 chromosomal Study population abnormalities (8.7%) were detected in this study. Nineteen of the cases (73.1%) were female and seven (26.9%) of them were male. A retrospective study was done in couples with recurrent first Fifteen individuals (57.7%) showed structural abnormalities and trimester abortions detected from 2012 to 2017 in Cumhuriyet eleven cases (42.3%) had numerical aberrations. In addition, there University Research Hospital. Recurrent miscarriage may be were 19 cases who have polymorphic variants or chromosomal defined as two or more failed clinical pregnancies that terminate variations. Chromosomal variations were predominant among the involuntarily before 20 weeks [9]. In this context, three hundred cases (Fig. 1). couples (600 individuals) who had two or more consecutive Among structural abnormalities in the group, reciprocal miscarriage were included in the study. The study group consisted translocations were detected in five cases (33.3%) and Robertso- of individuals aged 18 years and older with no systemic disease. nian translocations were found in four cases (26.7%). Three The types of recurrent miscarriage in the couples such as empty subjects (20%) had inversion anomaly. One of the inversions was gestational sac, missed abortions, chemical pregnancies and present on the Y chromosome. Duplications were detected in two spontaneous abortions were recorded for the study. The couples individuals (13.3%). There was a deletion of long arm of X were screened for multiple thrombophilic gene mutations. chromosome in one case (6.7%). Duplications and deletion were Hormone levels of women were measured and ultrasonography detected by aCGH (Cytoscan Optima, Thermo Fisher Scientific, and hysterosalpingography were used for the detection of possible USA). abnormalities of the genital tract. Blood tests for immunologic risk Eleven numerical chromosome aberrations were found in factors including antiphospholipid antibodies, antinuclear anti- couples with recurrent abortion and Turner mosaics were the most bodies and antithyroid antibodies were performed. Ectopic frequent numerical abnormalities. Five of the numerical anomalies pregnancies, women with anatomic abnormalities, endocrine (45.4%) were monosomy X mosaicism and in three cases (27.3%), disorder, autoimmune disease and men with sperm anomalies the combination of monosomy-trisomy X mosaicisms were were excluded from the study. This study was approved by detected. Two cases (18.2%) had the combination of mosaic Cumhuriyet University Ethics Committee. monosomy, trisomy and tetrasomy X. Only one of the numerical abnormalities (9.1%) was pentasomy X mosaicism. Mosaic cases Chromosome analysis were evaluated and confirmed by FISH probes (Vysis, Abbott, USA). We detected 19 chromosomal variations (heterochromatin and Metaphase chromosome preparations from the peripheral satellite increments and inv9) in our study group. Most frequent blood cultures were performed on the basis of standard polymorphic variant was inv9 with seven cases. Four individuals cytogenetic protocol. Peripheral blood samples in heparinized had 9qh + and the other heterochromatic variations were 1qh+, injectors were processed for karyotyping. Phytohaemagglutinin 16qh + and 15cenh + respectively. Five cases had satellit increments (0.1 mL) was used for the stimulation of cell proliferation and including 13, 14 or 15. chromosomes. cultured for 72 h at 37 C, in RPMI 1640 medium. Colchicine was added to the cultures. G-banded karyotyping was performed with Discussion trypsin–giemsa banding procedure. Twenty metaphases were scanned for each individual but for suspicious cases (mosaicism Spontaneous miscarriage is the most common complication in etc.) the practice was extended to fifty metaphases. pregnancy. A significant proportion of clinically defined pregnan- cies (about 15%) result in miscarriage [10]. Spontaneous abortion is FISH and aCGH often associated with chromosome anomalies. Approximately 40% of fetuses with an abnormal karyotype may inherit this anomaly Further molecular techniques, such as fluorescence in situ from one of their parents [11 ] and parental chromosomal hybridization (FISH) and array comparative genomic hybridization abnormalities can cause recurrent miscarriages. In this context, (aCGH) were used for detailed analysis. The first stage of the FISH karyotype analysis of couples is important to predict