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Mb 4Q22.2Q32.3 Duplication Due to Ovarian Germinal Mosaicism

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Mb 4Q22.2Q32.3 Duplication Due to Ovarian Germinal Mosaicism European Journal of Human Genetics (2010) 18, 882–888 & 2010 Macmillan Publishers Limited All rights reserved 1018-4813/10 www.nature.com/ejhg ARTICLE Recurrent 70.8 Mb 4q22.2q32.3 duplication due to ovarian germinal mosaicism Lucie Tosca*,1,2,3, Sophie Brisset1,2,4, Franc¸ois M Petit2,5, Laure Lecerf 6,7, Ghislaine Rousseau1,4,Ce´cile Bas2,3, Mireille Laroudie8, Marie-Laure Maurin1, Sylvie Tapia9, Olivier Picone10, Sophie Prevot8, Michel Goossens6,7, Philippe Labrune2,11,12 and Ge´rard Tachdjian1,2,3 A mosaicism is defined by the presence of two or more populations of cells with different genotypes in one individual. Chromosomal germinal mosaicism occurs in germ cells before the onset of meiosis. Previously, few studies have described germinal mosaicism. In this study, we report on two siblings who carried identical pure and direct interstitial 4q22.2q32.3 duplication. Procedure investigations included complete clinical description, conventional cytogenetic analysis, fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH) array experiments and microsatellite study searching for parental origin of the duplication. Microarray CGH and further FISH experiments with BAC clones showed the same 70.8 Mb direct duplication, dup(4)(q22.2q32.3). Molecular studies of the 4q duplication were consistent with maternal origin associated with mitotic or meiotic rearrangements. This structural chromosomal aberration was associated in both cases with increased nuchal translucency, growth retardation and dysmorphy. Cardiopathy and lung malformations were only evident in the first case. These clinical manifestations are similar to those previously reported in previous studies involving pure 4q trisomy of the same region, except for thumb and renal abnormalities that were not obvious in the presented cases. The amplified region included genes involved in neurological development (NEUROG2, MAB21L2, PCDH10/18 and GRIA2). The recurrent 4q duplication in these siblings is consistent with a maternal ovarian germinal mosaicism. This is the first description of germinal mosaicism for a large chromosomal duplication and highlights that genetic counselling for apparently de novo chromosome aberration should be undertaken with care. European Journal of Human Genetics (2010) 18, 882–888; doi:10.1038/ejhg.2010.46; published online 28 April 2010 Keywords: chromosome 4; duplication; recurrence; comparative genomic hybridization microarray; microsatellites; ovarian mosaicism INTRODUCTION comparative genomic hybridization (CGH) and due to ovarian A mosaicism is defined by the presence of two or more cell lines with germinal mosaicism. different genotypes in one individual, who has developed from a single fertilized egg.1 Different types of mosaicism exist, such as germinal MATERIALS AND METHODS mosaicism (restricted to germ cells) or tissue mosaicism. The most Clinical report common form of mosaicism detected through prenatal diagnosis The first pregnancy of a 24-year-old woman was marked by increased nuchal involves trisomies. Chromosomal germinal mosaicism is rare and translucency (4.5 mm) on first-trimester ultrasound examination. Parents were occurs in early germ cells (oogonia or spermatogonia) before the nonconsanguineous and both familial histories were unremarkable. Chorionic 1 onset of meiosis. These mitotic errors can be nondisjunctions villi were sampled for chromosomal analysis that showed a large long arm on (abnormality of chromosome number) or the production of structural chromosome 4. This chromosomal abnormality was confirmed on amniotic rearrangements (deletion, duplication, inversion, insertion or transloca- fluid analysis at 16 weeks. After genetic counselling and according to the French tion). Germinal mosaicism becomes evident when two siblings are born law, the pregnancy was terminated at 20 weeks of gestation. The female foetus with the same de novo chromosomal abnormality. Germinal mosaicism was hypotrophic. Weight, height, head circumference and biparietal diameter is independent of maternal age.2 Few studies in literature have described measurements were 221.6 g (o5th percentile), 15 cm (5th percentile), 25 cm (25th percentile) and 14 cm ( 25th percentile), respectively. Physical germinal mosaicism corresponding essentially to aneuploidies and o examination showed cranio-facial dysmorphism with elongated skull, large structural chromosomal aberrations including mostly isochromosomes 3–19 20 nose, long philtrum, large mouth with self-effacing cupidon arc, badly hemmed and deletions (for review, see Ro¨thlisberger and Kotzot ). ears and neck oedema (Supplementary Figure 1). Internal examination revealed In this study, we report two siblings carrying the same pure and pulmonary lobulation defect with two lobes in the right lung and a single one direct 4q22.2q32.3 duplication characterized using microarray in the left lung. Interauricular communication was also noted. The placenta was 1AP-HP, Histologie, Embryologie et Cytoge´ ne´ tique, Hoˆpital Antoine Be´cle` re, Clamart, France; 2Universite´ Paris Sud, Le Kremlin-Biceˆtre, France; 3INSERM U935, Villejuif, France; 4INSERM U782, Clamart, France; 5AP-HP, Biochimie, Hormonologie et Ge´ne´ tique, Hoˆ pital Antoine Be´ cle`re, Clamart, France; 6Universite´ Paris Val de Marne, Cre´teil, France; 7AP-HP, Plateforme de Ge´nomique IMRB 955, Hoˆ pital Henri Mondor, Cre´teil, France; 8AP-HP, Anatomie Pathologique, Hoˆ pital Antoine Be´cle` re, Clamart, France; 9Laboratoire Pasteur Cerba, Cytoge´ne´tique, Cergy Pontoise, France; 10AP-HP, Gyne´cologie Obste´trique, Hoˆpital Antoine Be´cle`re, Clamart, France; 11AP-HP, Pe´ diatrie, Hoˆpital Antoine Be´cle`re, Clamart, France; 12INSERM U948, Nantes, France *Correspondence: Dr L Tosca, Unite´ Fonctionnelle de Cytoge´ ne´tique, Service d’Histologie, Embryologie et Cytoge´ne´tique, Hoˆpital Antoine Be´cle`re, 157, rue de la Porte de Trivaux, Clamart Cedex 92141, France. Tel: +33 1 45 37 49 23; Fax: +33 1 45 37 42 07; E-mail: [email protected] Received 21 January 2010; revised 10 March 2010; accepted 17 March 2010; published online 28 April 2010 Duplication 4q and ovarian germinal mosaicism L Tosca et al 883 hypotrophic (86.8 g versus approximately 160 g for normal controls at the same Array comparative genomic hybridization (aCGH) term) but with normal aspect. The genomic imbalances of placenta and trophoblast cells were analysed by During the second pregnancy, increased nuchal translucency was again aCGH using 105K oligonucleotide arrays (Hu-105A, Agilent Technologies, detected (5 mm) on first-trimester ultrasound examination. Chorionic villi Massy, France). All array hybridizations were performed according to the were sampled and chromosomal analysis showed an abnormal long arm on manufacturer’s recommended protocols. In brief, 3 mg of genomic DNA was chromosome 4 similar to that observed in the first foetus. Regular foetus digested with AluI (5 units) and RsaI (5 units) for 2 h at 37 1C and fluorescently ultrasound examinations showed normal amniotic fluid volume and no labelled with the Agilent Genomic DNA labelling kit PLUS (Agilent Technol- malformations were detected in the foetus whose growth was normal. After ogies). A male or female human genomic DNA (Promega, Charbonnie`re, several discussions, both parents decided to continue the pregnancy. At 38 France) was used as reference. Experiments were conducted in dye-swap. weeks of gestation, a boy was vaginally delivered. His weight was 3040 g (o25th Cy5-dUTP patient DNA and its gender-matched reference labelled with percentile). Physical examination showed general hypotonia and facial dys- Cy3-dUTP were denatured and preannealed with Cot-1 DNA and Agilent morphism with narrow and horizontal palpebral fissures, prominent nose and blocking reagent before hybridization for 40 h at 20 r.p.m. in a 65 1C rotating moderate microretrognatia. At 6 weeks of age, his weight, height and head hybridization oven (Agilent Technologies). After washing, the slides were circumference were 4400 g (o25th percentile), 55.5 cm (o50th percentile) and scanned on an Agilent Microarray Scanner. Captured images were processed 37.5 cm (o25th percentile), respectively. The smile response was acquired, axial with Feature Extraction 9.1 (Agilent Technologies) software and data analysis tonus was correct and no other abnormality was specified. Facial dysmorphic was performed with CGH Analytics 3.5 (Agilent Technologies). Copy number features were stable. At the age of 6 months, the boy had no specific medical variations (CNVs) were considered significant if they were defined by three or treatment. His weight, height and head circumference were 7070 g (À1SD), more oligonucleotides spanning at least 50 Kb and contained at least one gene 64 cm (À1 SD) and 42 cm (À1 SD), respectively. He was able to catch close and were not identified in the Database of Genomic Variants. objects but not distant ones. He was able to burst into laughter. The seating position was not completely acquired and axial hypotonia was still present. At Fluorescence in situ hybridization (FISH) the age of 1 year, his weight, height and head circumference were 8900 g (À1 FISH analyses were performed on trophoblast cells and metaphase spreads SD), 70 cm (À1SD)and46cm(À1 SD), respectively. Axial hypotonia was still from both parental and propositus lymphocytes. The whole-chromosome marked, resulting in mild kyphosis when seating and in the absence of any painting (WCP) probe specific for chromosome 4 was used according to the possibility to stand
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