title: Clonal heterogeneity in childhood myelodysplastic syndromes--challenge for the detection of chromosomal imbalances by array-CGH.
abstract: To evaluate whether copy number alterations (CNAs) are present that may contribute to disease development and/or progression of childhood myelodysplastic syndromes (MDS), 36 pediatric MDS patients were analyzed using array-based comparative genome hybridization (aCGH). In addition to monosomy 7, the most frequent chromosome aberration in childhood MDS, novel recurrent CNAs were detected. They included a loss of 3p14.3-p12.3, which contains the putative tumor suppressor gene FHIT, a loss of 7p21.3-p15.3, a loss of 9q33.3-q34.3 (D184) and microdeletions in 17p11.2, 6q23 containing MYB, and 17p13 containing TP53. In this small patient cohort, patients without CNA, patients with monosomy 7 only and patients with one CNA in addition to monosomy 7 did not differ in their survival. As expected, all patients with complex karyotypes, including two patients with deletions of TP53, died. A challenge inherent to aCGH analysis of MDS is the low percentage of tumor cells. We evaluated several approaches to overcome this limitation. Genomic profiles from isolated granulocytes were of higher quality than those from bone marrow mononuclear cells. Decreased breakpoint calling stringency increased recognition of CNAs present in small clonal populations. However, further analysis using a custom-designed array showed that these CNAs often did not confirm the findings from 244k arrays. In contrast, constitutional CNVs were reliably detected on both arrays. Moreover, aCGH on amplified DNA from distinct myeloid clusters is a new approach to determine CNAs in small subpopulations. Our results clearly emphasize the need to verify array-CGH results by independent methods like FISH or quantitative PCR.
authors.affiliation: Institute of Cell and Molecular Pathology, Hannover Medical School, Hannover, Germany.
authors.forename: Inka
authors.initials: I
authors.lastname: Praulich
authors.forename: Marcel
authors.initials: M
authors.lastname: Tauscher
authors.forename: Gudrun
authors.initials: G
authors.lastname: Göhring
authors.forename: Stefanie
authors.initials: S
authors.lastname: Glaser
authors.forename: Winfried
authors.initials: W
authors.lastname: Hofmann
authors.forename: Simone
authors.initials: S
authors.lastname: Feurstein
authors.forename: Christian
authors.initials: C
authors.lastname: Flotho
authors.forename: Peter
authors.initials: P
authors.lastname: Lichter
authors.forename: Charlotte M
authors.initials: CM
authors.lastname: Niemeyer
authors.forename: Brigitte
authors.initials: B
authors.lastname: Schlegelberger
authors.forename: Doris
authors.initials: D
authors.lastname: Steinemann
mesh_terms: D000293:Adolescent; D000328:Adult; D001854:Bone Marrow Cells / Q000473:pathology; D002648:Child; D002675:Child, Preschool; D002869:Chromosome Aberrations; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics; D002897:Chromosomes, Human, Pair 7 / Q000235:genetics; D002899:Chromosomes, Human, Pair 9 / Q000235:genetics; D002999:Clone Cells; D055028:Comparative Genomic Hybridization; D005260:Female; D018628:Gene Dosage; D016147:Genes, Tumor Suppressor; D015894:Genome, Human; D006098:Granulocytes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007223:Infant; D008297:Male; D009190:Myelodysplastic Syndromes / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D055815:Young Adult