Chromosomal microarray analysis in foetuses with aberrant right subclavian artery

Idit Maya, 1,8

MD ,
Sarit Kahana, Josepha Yeshaya, 1
PhD ,
Tamar Tenne, Shiri Yacobson, 1
PhD ,
Ifaat Agmon-Fishman, 2
PhD , 1
MSc , 1
MSc ,
3 4 2,8
Lital Cohen-Vig, MD , Alex Levi, MD , Eyal Reinstein, MD, PhD , Lina Basel-Vanagaite, MD, PhD1,5,6,8, Reuven Sharony, MD2,7,8
1The Raphael Recanati Genetics Institute, Rabin Medical Center, Beilinson, Campus, Petah Tikva, Israel, 2The Genetics Institute, Meir Medical Center, Kfar Saba, Israel, 3Schneider
Children's Medical Center of Israel, Petah Tikva, Israel. 4Department of Cardiology, Meir Medical Center, Kfar Saba, Israel, 5Pediatric Genetics Unit, Schneider Children's Medical
Center of Israel, Petah Tikva, Israel, 6Felsenstein Medical Research Center, Rabin Medical Center, Petah Tikva, Israel, 7Department of Obstetrics and Gynecology, Meir Medical
Center, Kfar Saba, Israel, 8Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel

Abstract
Objectives: To evaluate the correlation between aberrant right subclavian artery (ARSA), with or without other ultrasound abnormality, and risk factors for aneuploidies and chromosomal microarray analysis (CMA) results. Methods and materials: This was a
multicentre study. Genetic analyses of foetuses diagnosed with ARSA were evaluated by CMA in the same laboratory. The clinical investigation included nuchal translucency, first and second trimester maternal serum screening, early and late second trimester foetal
anatomic scans and foetal echocardiogram. Comparative Genomic Hybridization (CGH) Microarray analysis or Single Nucleotide Polymorphism (SNP) Array technology was used for CMA. Results: CMA results were available for 63 foetuses diagnosed with ARSA.
No pathogenic variants were found among 36 foetuses with ARSA as an isolated finding. Additional ultrasound findings and/or risk factors for aneuploidies were present in 27 foetuses, of which 5 had pathogenic CMA results. Trisomy 21 (T21) was detected in a
foetus with echogenic intracardiac focus (EIF), 22q11 deletion was detected in a foetus with EIF and 1:230 maternal serum screening increased risk for T21, 22q11 duplication was detected in a foetus with hypoplastic right kidney and choroid plexus cysts, and
22q11 deletion in a foetus with right aortic arch and clubfoot. In the fifth foetus, both 22q11 deletion and 1q21 duplication were detected by CMA. In addition to ARSA, abnormal nuchal translucency (4 mm) and ventricular septal defect were also present.
Conclusions: The results suggest that in foetuses with ARSA as an isolated finding, an invasive procedure for CMA testing is not indicated. However, CMA is recommended when additional ultrasound abnormalities or risk factors for aneuploidies are detected. The
chromosomal findings in 4 of the 5 abnormal cases would not have been detected by standard fetal chromosomal testing

Introduction Results References Tables

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Methods
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CPC 23,756,844 n
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26. Volpe P, Marasini M, Caruso G,Marzullo A, Buonadonna AL, Arciprete P, Di Paolo S, Volpe G and Gentile M. 22q11 deletions in 4 NT 4 mm, 1q21.1- Duplication 0.9 chr1:146,501 1q21
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translucency and ventricular septal defect (VSD) was diagnosed 752-757.
27. Chaoui, R., Heling, K.-S., Sarut Lopez, A., Thiel, G. and Karl, K. The thymic–thoracic ratio in fetal heart defects: a simple way to
VSD q21.2 ,348- duplicatio
The study cohort included 63 foetuses that were diagnosed with with 22q11 deletion (2.7 Mb, chr22:18,844,632-21,536,104) and identify fetuses at high risk for microdeletion 22q11. Ultrasound Obstet Gynecol 2011, 37, 397–403.

ARSA from July 2013 through September 2015, based on US gain of chromosome 1q21 (0.9 Mb, chr1:146,501,348-147,398,560). Table 1: Indications for CMA in foetuses with ARSA and 22q11.21 Deletion 2.7 147,398,560 n
findings. All women underwent genetic counselling and signed an In this group, 87% of the women were younger than 35 and 64% of the results according to additional findings chr22:18,844 DiGeorge
infor med consent that explained the possible pathogenic the foetuses were female. A detailed summary of the 5 affected
significance of this finding. This was followed by amniocentesis and cases is presented in Table 2. There was no correlation between ,632- /VCF
Group Indication Number Pathogenic CMA findings
fetal CMA testing. The cohort was classified into three groups: A) ARSA and foetal gender..The likelihood ratio of positive results 21,536,104
isolated ARSA; B) ARSA and soft ultrasonographic markers for T21 (LR+) in cases of complicated ARSA, were 26.81 for T21 and for invasive of cases cases (%) [95%CI]
5 Right aortic 22q11.21 Deletion 2.9 chr22: DiGeorge
(e.g., echogenic intracardiac focus (EIF) and single umbilical artery 107.33 for 22q11 deletion syndrome
(SUA)), and/or screen positive and/or advanced maternal age diagnosis arch, clubfoot 17,397,529- /VCF
(AMA); and C) ARSA and major ultrasonographic anomalie This was Conclusion A ARSA – 0 (0%) 20,311,734
a multicenter study. Thus, although ARSA detection protocols were
36
similar
, they were unavailable, as were perinatal results. The genetic In conclusion, we present CMA results from a large cohort of isolated
analysis was done in the same laboratory and the protocol is foetuses with ARSA -- either isolated or complicated. Our results AMA – advanced maternal age; CPC – choroid plexus cyst; EIF – echogenic
detailed in the appendix. ARSA detection - The detection of ARSA showed that cases of isolated ARSA do not have increased risk of B ARSA and 2 (20%)* [5.7%-51%] intra cardiac focus; MSS – maternal serum screening; NT – nuchal
translucency; VSD – ventricular septal defect; VCF – Velocardiofacial
was performed in the three vessels and trachea axial view. Most aneuploidies. These results are consistent with previous studies
cases were detected initially by an obstetrician performing fetal suggesting that there is insufficient evidence to discuss CMA soft signs for 10
anatomical scan. All evaluations were performed either abnormalities in cases of isolated ARSA when the mother is at low-
aneuploidies
transvaginally with a 5–9 MHz probe or transabdominally with a 4–8 risk for T21. In such cases, detailed US scans including cardiac
MHz probe (Voluson E6 or E8, GE Medical Systems, Zipf, Austria), evaluation should be carried out in search of additional T21 and C ARSA and 3 (18%)† [6.2%-41%]
as appropriate. The studies were followed by fetal echocardiography
performed after 20 weeks-gestation or a few days after the first
22q11 deletion markers (e.g. thymic aplasia27). Based on the
literature reviewed and our experience that CMA detection rate in major US 17
Acknowledgements
examination by a sonographer specializing in fetal complicated ARSA is approximately 20%, invasive procedures, such
echocardiography. These studies were performed transabdominally anomalies We would like to acknowledge the following for
as amniocentesis are indicated. In these cases, rapid chromosomal
with a 4–8-MHz probe (iE33 echo system, Philips medical; Andover, their help in making this research possible:
evaluation, e.g., by FISH (fluorescent in situ hybridization) might be Total 5 (8%) [3.4%-17.3%]
63 Faye Schriber. MSc; Nava Yelin, MSc
MA). DNA extraction and Chromosomal microarray analysis (CMA) - considered prior to full CMA study. Future large scale studies are Trisomy 21; 22q11 deletion; †22q11 deletion; 22q11 deletion+ 1q21
These procedures are detailed in the appendix needed to analyse the correlations of DS to the entire area of duplication; 22q11 deletion
cardiovascular anomalies, especially ARSA, whether isolated or not. ARSA – ab*errant right subclavian artery; CMA – chromosomal microarray
analysis; US – ultrasound