Open Access

Molecular cytogenetic analysis and clinical manifestations of a case with de novo mosaic ring chromosome 7

  • Li-Ping Tsai1, 2,
  • Kuei-Fang Lee3,
  • Jye-Siung Fang3, 4 and
  • Ingrid Y Liu4Email author
Contributed equally
Molecular Cytogenetics20114:5

DOI: 10.1186/1755-8166-4-5

Received: 14 September 2010

Accepted: 8 February 2011

Published: 8 February 2011

Abstract

Aim

Clinical and molecular cytogenetic investigations of a newborn girl exhibiting facial dysmorphism with developmental delay.

Methods

Phenotypic evaluation was first applied to examine the proband's developmental status. Computed tomography and colour transcranial Doppler were used then to investigate her brain structure and function. Subsequently, chromosomal abnormalities were examined by karyotyping and fluorescent in situ hybridization was performed to investigate size of fragments lost at the two distal ends of the ring chromosome 7. In addition, multicolour banding was applied to rule out structural rearrangement occurs in between the ring chromosome 7.

Results

The proband was born with mosaic supernumerary ring chromosome 7, without a normal karyotype detected in the peripheral blood lymphocytes. The distal arm of chromosome 7p (at least 255 kb from the telomere) was part of an extra ring chromosome 7. In addition, the distal arm of 7q, at least 8 kb from the telomere, was missing. There was no other chromosomal rearrangement detected by multicolour banding.

Interpretation

This is the 19th reported case of complete ring chromosome 7 mosaicism and the first survived case with mosaic supernumerary ring 7 without a normal karyotype detected in the peripheral lymphocytes.

Introduction

Ring chromosome 7 is a rare chromosome anomaly that leads to variable phenotypes. The first two cases were described by Zackai et al in 1973 [1]. A total of 18 cases with complete ring chromosome 7 have been reported to date worldwide [117]. Ring chromosomes are often unstable during mitosis; therefore, it is common to find a ring chromosome in only a portion of all cells analyzed (mosaicism). The other cells in an individual, with a ring chromosome, are usually monosomic or demonstrate partial-trisomy with a small ring [18]. Here we present the 19th case of complete ring chromosome 7 mosaicism. This is the first survived case with complete supernumerary ring 7, without a normal karyotype detected in the peripheral lymphocytes. The phenotypic expression of patients with ring chromosome 7 is variable; most patients demonstrate developmental delay, mental retardation, microcephaly, and dermatological abnormalities including cafe-au-lait spots, nevus flammeus and dark pigmented nevi (for review, please see [7]). The variable phenotypes may result from the variable size of the deleted chromosomes at the terminal segments, ring instability, and/or the level of mosaicism. The case presented here had microcephaly, hypotelorism, choanal stenosis, and speech delay, without any dermatological abnormalities detected to date.

Methods

Patient and Clinical examination

A female child with microcephaly (Figure 1 A and 1B, pictures taken at one year and 10 months of age) and respiratory distress was referred to the genetic counselling at the department of Pediatrics at Tzu Chi General Hospital in Taipei County, Taiwan. The proband was the first child of non-consanguineous parents. The Vietnamese mother was 23 at delivery, and the Taiwanese father was in his 40 s. The proband was born by caesarean section with a birth weight of 2410 gm, length 42 cm and head circumference of 30 cm, all fall below the third percentile for gestational age. Facial dysmorphism was impressed, including hypotelorism, midface hypoplasia and high arch palate. Choanal stenosis was suspected because of noise nasal breathing since birth. Physical examination also detected subcutaneous syndactyly of 3rd and 4th toes. Computed tomography (CT) and transcranial Doppler were directed due to microcephaly. CT of the brain showed mild stepage, widening of the upper part of the right lambdoid suture and mild scalp thickening along the course of the right lambdoid suture. The pediatric brain echo revealed normal ventricles and choroid plexus pulsation; however, a hypoechoic cyst-like lesion was noted next to the quadrigeminal plate. Speech delay and psychomotor retardation was noted during clinical evaluation at thirteen months old. The body weight of 5.4 Kg, height of 61 cm and head circumference of 35.2 cm still fell far below the third percentile on growth curve.
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Figure 1

Pictures taken from the front (A) and from the side (B) of the proband at her age of one year and 10 months old. Facial dysmorphism including hypotelorism, midface hypoplasia and high arch palate were noted. Choanal stenosis was suspected because of noise nasal breathing since birth.

Metaphases preparation and florescent in situ hybridization (FISH)

Metaphases slides were prepared from lymphocyte cultures and G-banded for karyotyping using a standard protocol [19]. Five metaphase slides then were incubated in 2 × SSC (pH7.0) prewarmed to 37°C for 30 min for FISH analysis. After that, slides were dehydrated sequentially in a 70%, 85% and 100% ethanol series for 2 min each and then air dried. The metaphase slides were next denatured for 5 min in denaturant solution (70% formamide/2 × SSC) in a 73°C water bath inside of a coplin jar, and then dehydrated in serial ethanol for 2 min each. Subsequently, 10 μl of denatured probes (G31340 for chromosome 7q and G31341 for 7p, Cytocell, Ltd, Cambridge, UK) were added to the metaphase slides and then covered with a cover glass; the slides were placed in a prewarmed humidified box and incubated overnight at 42°C for hybridization. The next day, the cover glasses were removed immediately and the slides were washed with 0.4 × SSC/0.3% NP40 three times for 2 min each, and then air dried in the dark. Hybridization areas were counterstained with 20 μl DAPI (Vysis Inc, IL, USA) and then examined under a fluorescence microscope.

Multicolor banding

Two metaphase slides were prepared for multicolour banding FISH analysis, Multicolor banding FISH probes for chromosome 7 (XCyte 7 24 μl D-0207-024-MC, XCyte 7 24 μl D-0207-024-MC) were hybridized to metaphases using standard FISH protocol described above and viewed under the fluorescent microscope (Axioskop 40, Carl Zeiss Inc., Germany).

Results

Chromosome analysis of the peripheral lymphocytes revealed a mosaic karyotype. Out of 100 cells analyzed, a 46,XX,r(7)(p22q36) (Figure 2A) complement was observed in 75 cells (75%); 45,XX, -7 (Figure 2B) in 12 cells (12%); 47,XX,r(7)(P22q36),+r(7)(p22q36) (Figure 2C) in five cells (5%); 46,XX,dic r(7;7)(p22q36;p22q36)(Figure 2D) in four cells (4%); 47,XX,r(7), +dicr(7;7)(p22q36;p22q36), and 47,XX,r(7),+inv(7)(p11.1-p22::p36-q11.1) in two cells (2%). A total of 13% of the examined cells revealed a supernumerary ring 7. Skin fibroblast analysis was planned but the family did not return for further evaluations. A subtelomere specific probe was hybridized to metaphase cells by fluorescent in situ hybridization (FISH) to determine how much of the distal region of chromosome 7 remained intact in the ring chromosome 7. The results showed that the distal segment of chromosome 7p (at least 255 kb from telomere) appeared to be present in all of the ring chromosome 7 fragments (Figure 3A-C). However, the distal segment of chromosome 7q (at least 8 kb from telomere) was missing (Figure 3D).
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Figure 2

Mosaic karyotypes of the proband: Out of 100 analyzed cells, 46, XX,r(7) (p22q36) (A) was detected in 75 cells (75%); 45,XX, -7 (B) in 12 cells (12%); 47,XX,r(7)(p22q36),+ r(7)(p22q36) (C) in five cells (5%); and 46, XX, dic r (7;7)(p22q36;p22q36)(D) in four cells (4%).

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Figure 3

Flourescent In Situ Hybridization (FISH) using 7p and 7q sub-telomere probes: FISH with 7pter probe indicates subtelomeric region at least 255 kb from telomere remained intact in all rings (A, B and C). However, the 7qter probe indicates that at least 8 kb from telomere was missing in the ring (D).

Multicolour banding was performed to determine whether a micro-rearrangement occurred in the ring chromosome 7. Multicolor chromosome 7 banding probes that specifically detect various regions along chromosome 7 were hybridized to metaphases and viewed under the fluorescent microscope. No chromosome rearrangement was detected (Figure 4).
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Figure 4

Multicolor banding of normal and ring chromosome 7: Multicolor banding probes that specifically detect various regions along chromosome 7 were hybridized to metaphases; no rearrangement was observed in normal ring 7 chromosomes.

Discussion

A ring chromosome is formed by telomere-to-telomere fusion with minimal or no loss of genomic material. Ring chromosomes are often unstable during mitosis; as a result, it is common to find a ring chromosome in only a portion of cells (mosaicism). The other cells in the individual are usually monosomic or demonstrate partial-trisomy. Some cells lose the ring chromosome and some cells maintain a supernumerary ring chromosome in which the ring is usually small and composed mostly of the centromere and a small amount of euchromatic material [19].

The karyotype of the patient presented in this report was complicated and unique. There was no normal karyotype detected in the peripheral blood lymphocytes. Monosomy 7, one complete ring, supernumerary large rings, and duplicated rings of chromosome 7, were detected. Most previous reported cases of supernumerary ring chromosome 7 showed a majority of cells with a normal karyotype, in addition to some cells with a partial trisomy containing small rings [2026]. Here we present a newborn girl that survived with a mosaic karyotype with a complete supernumerary ring chromosome 7 and a few monosomy 7 cells, without any cell with normal karyotype detected in peripheral lymphocytes. All large rings have an intact 7p G31341 region with the 7q telomere absent (8 kb from telomere). According to the human chromosome 7 view map (http://www.ncbi.nlm.nih.gov) of the National Centre for Biotechnology Information (NCBI), there are no functional genes located within the missing distal region of 7q. There might be a microdeletion at the 7p telomere; however, no commercial probe available yet to detect a missing segment in this region.

In a previous report of two cases with complete trisomy ring 7 the infants were stillborn [27, 28]. In the present case, a majority of cells (75%) were composed of 46, XX, r(7), and all of the large rings appeared to be stable; this is likely associated with the patient's survival. However, it is also possible that other tissues, beside the peripheral lymphocytes, had a normal karyotype; but no other tissue samples were available for analysis. The influence of gene dosage effects resulting from monosomy chromosome 7 (12%) and supernumerary ring chromosome 7 (13%) were likely the cause of the abnormal clinical characteristics in the patient reported here. We also assumed uniparental disomy (UPD) of chromosome 7 was associated with the proband's clinical manifestation, however, further examination for UPD was not possible due to relocation of this family.

The clinical expression of ring chromosome 7 is variable. Nevertheless, most cases share some characteristics such as: mental retardation, growth deficiency, mild microcephaly, facial asymmetry, hypertelorism, abnormal palpebral fissures, small ears, limb and skeletal anomalies, and skin lesions, including nevus flammeus, dark pigmented nevi and café-au-lait spots [7, 10]. Our patient had some of these findings e.g. growth retardation, microcephally. However, no pigmented nevi or café-au-lait spots were observed at the time of evaluation. In addition, speech delay was noted. These findings suggest that some of the genes located within the affected region of chromosome 7 are associated with the cognitive abilities needed for speech development and might illustrate a dosage-dependent pattern.

In summary, we present the first case of survival with a mosaic and supernumerary complete ring chromosome 7, without a normal karyotype detected in peripheral blood lymphocytes. The high percentage (75%) of cells with a stable 46,XX, r(7), the absence of a macro-deletion and rearrangement within the ring chromosome 7 are likely the major reasons accounting for the viability of this patient.

Consent

Written informed consent was obtained from the parents of the patient for publication of this case report and accompanying images.

Notes

Declarations

Acknowledgements

This study was supported by Ministry of Economic Affairs, Taiwan, grant 98-EC-17-A-19-S2-0110. We are grateful to Dr. Ming-Liang Lee for helpful comments on the manuscript.

Authors’ Affiliations

(1)
Department of Pediatrics, Buddhist Tzu Chi General Hospital
(2)
Department of Medicine, Tzu Chi University
(3)
Laboratory for Cytogenetics, Centre for Genetic Counselling, Buddhist Tzu Chi Hospital
(4)
Department of Molecular Biology and Human Genetics, Tzu Chi University

References

  1. Zackai EH, Breg WR: Ring chromosome 7 with variable phenotypic expression. Cytogenet Cell Genet 1973,12(1):40–48. 10.1159/000130436PubMedView ArticleGoogle Scholar
  2. Barros LP PA, Brunoni D, Andrade JAD: Cromosomo 7 emanel-relato de um caso. Cienc Cult 1986, 38: 849.Google Scholar
  3. Biesecker LG C, Glover TW: Severe anomalies associated with ring chromosome 7. American Journal of Medical Genetics 1991, 40: 429–431. 10.1002/ajmg.1320400410PubMedView ArticleGoogle Scholar
  4. Caramia GM, Baroncini A, Osimani P, Forabosco A: Ring chromosome 7: report of the fifth case. Eur J Pediatr 1990,149(7):475–476. 10.1007/BF01959398PubMedView ArticleGoogle Scholar
  5. Ceballos-Quintal JM, Pinto-Escalante D, Castillo-Zapata H, Canto-Herrera J: Severe congenital hypoacusia in a patient with mosaic ring chromosome 7. Rev Biomed 1999, 10: 235–239.Google Scholar
  6. DeLozier CD, Theintz G, Sizonenko P, Engel E: A fourth case of ring chromosome 7. Clin Genet 1982,22(2):90–98. 10.1111/j.1399-0004.1982.tb01419.xPubMedView ArticleGoogle Scholar
  7. Kaur A, Dhillon S, Garg PD, Singh JR: Ring chromosome 7 in an Indian woman. J Intellect Dev Disabil 2008,33(1):87–94. 10.1080/13668250701829829PubMedView ArticleGoogle Scholar
  8. Kohyama J, Watanabe S, Nakajima M, Suzumura H, Ishikawa T, Ishikawa K, Saito F, Fukuda C: Ring chromosome 7: report of a case. Acta Paediatr Jpn 1988, 30: 517–519.PubMedView ArticleGoogle Scholar
  9. Koiffmann CP, Diament A, de Souza DH, Wajntal A: Ring chromosome 7 in a man with multiple congenital anomalies and mental retardation. J Med Genet 1990, 27: 462–464. 10.1136/jmg.27.7.462PubMed CentralPubMedView ArticleGoogle Scholar
  10. Mehraein Y, Ehlhardt S, Wagner A, Gottert E, Tilgen W, Zang KD, Dill-Muller D: Somatic mosaicism of chromosome 7 in a highly proliferating melanocytic congenital naevus in a ring chromosome 7 patient. Am J Med Genet A 2004,131(2):179–185. 10.1002/ajmg.a.30370PubMedView ArticleGoogle Scholar
  11. Nakano S, Miyamoto N: A ring C7 chromosome in a mentally and physically retarded male with various somatic abnormalities. Jinrui Idengaku Zasshi 1977, 22: 33–41. 10.1007/BF01908283PubMedView ArticleGoogle Scholar
  12. Rodriguez L, Sanchis A, Villa A, Canovas A, Peris S, Estivalis M, Pons S, Martinez-Frias ML: Ring chromosome 7 and sacral agenesis. Am J Med Genet 2000,94(1):52–58. 10.1002/1096-8628(20000904)94:1<52::AID-AJMG11>3.0.CO;2-QPubMedView ArticleGoogle Scholar
  13. Sawyer JR, Lukacs JL, Hassed SJ, Arnold GL, Mitchell HF, Muenke M: Sub-band deletion of 7q36.3 in a patient with ring chromosome 7: association with holoprosencephaly. Am J Med Genet 1996, 65: 113–116. 10.1002/(SICI)1096-8628(19961016)65:2<113::AID-AJMG6>3.0.CO;2-TPubMedView ArticleGoogle Scholar
  14. Tsai PY LY, Chang CH, Chang FM, Kuo PL: Ring chromosome 7 presenting with intrauterine growth restriction and multiple anomalies. Taiwanese Journal Obstet Gynecol 2005,44(3):297299.View ArticleGoogle Scholar
  15. Tsukamoto H, Sakai N, Taniike M, Nakatsukasa M, Yoshiwara W, Sakamoto H, Fujimura H, Inui K, Okada S: Case of ring chromosome 7: the first report of neuropathological findings. Am J Med Genet 1993,46(6):632–635. 10.1002/ajmg.1320460606PubMedView ArticleGoogle Scholar
  16. Vermeesch JR, Baten E, Fryns JP, Devriendt K: Ring syndrome caused by ring chromosome 7 without loss of subtelomeric sequences. Clin Genet 2002,62(5):415–417. 10.1034/j.1399-0004.2002.620511.xPubMedView ArticleGoogle Scholar
  17. Wahlstrom J, Bjarnason R, Rosdahl I, Albertsson-Wikland K: Boy with a ring 7 chromosome: a case report with special reference to dermatological findings. Acta Paediatr 1996,85(10):1256–1260. 10.1111/j.1651-2227.1996.tb18243.xPubMedView ArticleGoogle Scholar
  18. Mueller RF, Young ID: Chromosomes and Cell Division. In Emery's Elements of Medical Genetics. Volume Chapter 3. 11th edition. Churchill Livingstone Elsevier; 2007.Google Scholar
  19. Barch MJ: The ACT Cytogenetics Laboratory Manual. 2nd edition. Raven Press, New York; 1991.Google Scholar
  20. Bertini V, Valetto A, Uccelli A, Bonuccelli A, Tarantino E, Taddeucci G, Simi P: Molecular cytogenetic characterization of a de novo mosaic supernumerary ring chromosome 7: report of a new patient. Am J Med Genet A 2008,146A(22):2955–2959. 10.1002/ajmg.a.32528PubMedView ArticleGoogle Scholar
  21. Chantot-Bastaraud S, Muti C, Pipiras E, Routon MC, Roubergue A, Burglen L, Siffroi JP, Simon-Bouy B: Clinical findings and cytogenetic analysis of small supernumerary ring chromosomes 7: report of two new cases. Ann Genet 2004,47(3):241–249.PubMedView ArticleGoogle Scholar
  22. Combi R, Sala E, Villa N, Crosti F, Beccaria L, Cogliardi A: Tenchini ML, Dalpra L. Maternal heterodisomy/isodisomy and paternal supernumerary ring of chromosome 7 in a child with Silver-Russell syndrome. Clin Dysmorphol 2008,17(1):35–39. 10.1097/MCD.0b013e328235a587PubMedView ArticleGoogle Scholar
  23. Lichtenbelt KD, Hochstenbach R, van Dam WM, Eleveld MJ, Poot M, Beemer FA: Supernumerary ring chromosome 7 mosaicism: case report, investigation of the gene content, and delineation of the phenotype. Am J Med Genet A 2005,132A(1):93–100. 10.1002/ajmg.a.30408PubMedView ArticleGoogle Scholar
  24. Tan-Sindhunata G, Castedo S, Leegte B, Mulder I, vd Veen AY, vd Hout AH, Wiersma TJ, van Essen AJ: Molecular cytogenetic characterization of a small, familial supernumerary ring chromosome 7 associated with mental retardation and an abnormal phenotype. Am J Med Genet 2000,92(2):147–152. 10.1002/(SICI)1096-8628(20000515)92:2<147::AID-AJMG13>3.0.CO;2-IPubMedView ArticleGoogle Scholar
  25. Velagaleti GV, Jalal SM, Kukolich MK, Lockhart LH, Tonk VS: De novo supernumerary ring chromosome 7: first report of a non-mosaic patient and review of the literature. Clin Genet 2002,61(3):202–206. 10.1034/j.1399-0004.2002.610306.xPubMedView ArticleGoogle Scholar
  26. von Beust G, Sauter SM, Liehr T, Burfeind P, Bartels I, Starke H, von Eggeling F, Zoll B: Molecular cytogenetic characterization of a de novo supernumerary ring chromosome 7 resulting in partial trisomy, tetrasomy, and hexasomy in a child with dysmorphic signs, congenital heart defect, and developmental delay. Am J Med Genet A 2005,137(1):59–64.PubMedView ArticleGoogle Scholar
  27. Verp MS, Amarose AP: Chromosomal mosaicism of trisomy 7 restricted to chorionic villi. Am J Med Genet 1987,28(1):221. 10.1002/ajmg.1320280131PubMedView ArticleGoogle Scholar
  28. Yunis E, Ramirez E, Uribe JG: Full trisomy 7 and Potter syndrome. Hum Genet 1980,54(1):13–18. 10.1007/BF00279043PubMedView ArticleGoogle Scholar

Copyright

© Tsai et al; licensee BioMed Central Ltd. 2011

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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