title: Interstitial deletions at the long arm of chromosome 13 may be as common as monosomies in multiple myeloma. A genotypic study.
abstract: OBJECTIVE
  Deletions at the long arm of chromosome 13, mostly at the q14, and monosomy of chromosome 13 are described to be common in multiple myeloma (MM). 13q- has been associated with an adverse outcome and it has been proposed as one of the most important prognostic factors for MM patients. Deletions of 13q14 are rare in monoclonal gammopathy of undetermined significance (MGUS) and are thus believed to be associated with the development of the full myeloma phenotype.
  
  
  METHODS
  A genotyping analysis on purified neoplastic plasma cells was performed on 14 consecutive MM cases to determine the minimally deleted region at chromosome 13. Freshly obtained bone marrow was analyzed by flow cytometry in order to establish the percentage of plasma cell infiltration (CD38+BB4+CD56+/-CD19-). Neoplastic enrichment was carried out using BB4 coated immunomagnetic beads. This method allowed us to monitor the enrichment process. DNA obtained from the enriched neoplastic population and DNA from the clean fraction was amplified using combination sets of chromosomes 12 and 13. Amplimers were run on acrylamide gels, analyzed by automatic fluorescence quantification, and their size determined using the software programs Genescan and Genotyper.
  
  
  RESULTS
  In 11 patients electropherograms were suggestive of loss of heterozygosity (LOH) for polymorphic markers located at the long arm of chromosome 13. Four patients showed monosomy and 7 had interstitial deletions in the telomeric region. LOH was not evidenced at chromosome 12 in any sample. The minimal region with deletion was defined by the markers D13S159 (13q32.2, centromeric) and D13S1267 (13q32.3, telomeric). A gene with a potentially pathogenic role may be located in this very small region. Three patients did not show LOH at chromosome 13 by genotypic analysis; however, in one of these, proximal deletion at the long arm of chromosome 13 (13q2.1-2.2) was demonstrated by comparative genomic hybridization (CGH).
  
  
  CONCLUSIONS
  These findings suggest that interstitial deletions of the long arm of chromosome 13 may be more common than previously recognized. The methodologic approach reported in this work may simplify LOH analysis in MM patients. The potential uses of genotyping analysis in risk stratification remain to be investigated.
authors.affiliation: Laboratori d Hematologia, Hospital de la Santa Creu i Sant Pau, Avda Sant Antoni M. Claret 167, 08025 Barcelona, Spain. jnomdedeu@hsp.santpau.es
authors.forename: Josep F
authors.initials: JF
authors.lastname: Nomdedéu
authors.forename: Adriana
authors.initials: A
authors.lastname: Lasa
authors.forename: Josep
authors.initials: J
authors.lastname: Ubeda
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Saglio
authors.forename: Mar
authors.initials: M
authors.lastname: Bellido
authors.forename: Sílvia
authors.initials: S
authors.lastname: Casas
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Carnicer
authors.forename: Anna
authors.initials: A
authors.lastname: Aventín
authors.forename: Anna
authors.initials: A
authors.lastname: Sureda
authors.forename: Jorge
authors.initials: J
authors.lastname: Sierra
authors.forename: Montserrat
authors.initials: M
authors.lastname: Baiget
mesh_terms: D000328:Adult; D000368:Aged; D001938:Brazil; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D002882:Chromosomes, Human, Pair 13 / Q000235:genetics* / Q000648:ultrastructure; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005819:Genetic Markers; D005838:Genotype; D006801:Humans; D016130:Immunophenotyping; D019656:Loss of Heterozygosity; D008297:Male; D018895:Microsatellite Repeats; D008875:Middle Aged; D009006:Monosomy; D009101:Multiple Myeloma / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
chemical_list: D004273:DNA, Neoplasm; D005819:Genetic Markers