title: Detection of genomic imbalances in microdissected Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma by array-based comparative genomic hybridization.
abstract: BACKGROUND
  Cytogenetic analysis of classical Hodgkin's lymphoma is limited by the low content of the neoplastic Hodgkin-Reed-Sternberg cells in the affected tissues. However, available cytogenetic data point to an extreme karyotype complexity. To obtain insights into chromosomal imbalances in classical Hodgkin's lymphoma, we applied array-based comparative genomic hybridization (array comparative genomic hybridization) using DNA from microdissected Hodgkin-Reed-Sternberg cells.
  
  
  METHODS
  To avoid biases introduced by DNA amplification for array comparative genomic hybridization, cHL cases rich in Hodgkin-Reed-Sternberg cells were selected. DNA obtained from approximately 100,000 microdissected Hodgkin-Reed-Sternberg cells of each of ten classical Hodgkin's lymphoma cases was hybridized onto commercial 105 K oligonucleotide comparative genomic hybridization microarrays. Selected imbalances were confirmed by interphase cytogenetics and quantitative polymerase chain reaction analysis and further studied in an independent series of classical Hodgkin's lymphoma.
  
  
  RESULTS
  Gains identified in at least five cHL affected 2p12-16, 5q15-23, 6p22, 8q13, 8q24, 9p21-24, 9q34, 12q13-14, 17q12, 19p13, 19q13 and 20q11 whereas losses recurrent in at least five cases involved Xp21, 6q23-24 and 13q22. Copy number changes of selected genes and a small deletion (156 kb) of the CDKN2B (p15) gene were confirmed by interphase cytogenetics and polymerase chain reaction analysis, respectively. Several gained regions included genes constitutively expressed in cHL. Among these, gains of STAT6 (12q13), NOTCH1 (9q34) and JUNB (19p13) were present in additional cHL with the usual low Hodgkin-Reed-Sternberg cell content.
  
  
  CONCLUSIONS
  The present study demonstrates that array comparative genomic hybridization of microdissected Hodgkin-Reed-Sternberg cells is suitable for identifying and characterizing chromosomal imbalances. Regions affected by genomic changes in Hodgkin-Reed-Sternberg cells recurrently include genes constitutively expressed in cHL.
authors.affiliation: Senckenberg Institute of Pathology, University of Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany. s.hartmann@em.uni-frankfurt.de
authors.forename: Sylvia
authors.initials: S
authors.lastname: Hartmann
authors.forename: José I
authors.initials: JI
authors.lastname: Martin-Subero
authors.forename: Stefan
authors.initials: S
authors.lastname: Gesk
authors.forename: Julia
authors.initials: J
authors.lastname: Hüsken
authors.forename: Maciej
authors.initials: M
authors.lastname: Giefing
authors.forename: Inga
authors.initials: I
authors.lastname: Nagel
authors.forename: Jennifer
authors.initials: J
authors.lastname: Riemke
authors.forename: Andreas
authors.initials: A
authors.lastname: Chott
authors.forename: Wolfram
authors.initials: W
authors.lastname: Klapper
authors.forename: Marie
authors.initials: M
authors.lastname: Parrens
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Merlio
authors.forename: Ralf
authors.initials: R
authors.lastname: Küppers
authors.forename: Andreas
authors.initials: A
authors.lastname: Bräuninger
authors.forename: Reiner
authors.initials: R
authors.lastname: Siebert
authors.forename: Martin-Leo
authors.initials: ML
authors.lastname: Hansmann
mesh_terms: D055028:Comparative Genomic Hybridization; D015894:Genome, Human / Q000235:genetics*; D006689:Hodgkin Disease / Q000145:classification / Q000235:genetics* / Q000601:surgery; D006801:Humans; D042282:Microdissection; D020411:Oligonucleotide Array Sequence Analysis; D016539:Reed-Sternberg Cells / Q000378:metabolism*