Split-signal FISH for detection of chromosome aberrations in acute lymphoblastic leukemia.
Chromosome aberrations are frequently observed in precursor-B-acute lymphoblastic leukemias (ALL) and T-cell acute lymphoblastic leukemias (T-ALL). These translocations can form leukemia-specific chimeric fusion proteins or they can deregulate expression of an (onco)gene, resulting in aberrant expression or overexpression. Detection of chromosome aberrations is an important tool for risk classification. We developed rapid and sensitive split-signal fluorescent in situ hybridization (FISH) assays for six of the most frequent chromosome aberrations in precursor-B-ALL and T-ALL. The split-signal FISH approach uses two differentially labeled probes, located in one gene at opposite sites of the breakpoint region. Probe sets were developed for the genes TCF3 (E2A) at 19p13, MLL at 11q23, ETV6 at 12p13, BCR at 22q11, SIL-TAL1 at 1q32 and TLX3 (HOX11L2) at 5q35. In normal karyotypes, two colocalized green/red signals are visible, but a translocation results in a split of one of the colocalized signals. Split-signal FISH has three main advantages over the classical fusion-signal FISH approach, which uses two labeled probes located in two genes. First, the detection of a chromosome aberration is independent of the involved partner gene. Second, split-signal FISH allows the identification of the partner gene or chromosome region if metaphase spreads are present, and finally it reduces false-positivity.
van der Burg
M
M
Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
Poulsen
T S
TS
Hunger
S P
SP
Beverloo
H B
HB
Smit
E M E
EM
Vang-Nielsen
K
K
Langerak
A W
AW
van Dongen
J J M
JJ
D051792:Basic Helix-Loop-Helix Proteins; D002869:Chromosome Aberrations; D004268:DNA-Binding Proteins / Q000235:genetics; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D051788:Myeloid-Lymphoid Leukemia Protein; D015513:Oncogene Proteins / Q000235:genetics; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics*; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins / Q000235:genetics; D051562:Proto-Oncogene Proteins c-bcr; D050783:Proto-Oncogene Proteins c-ets; D011519:Proto-Oncogenes; D012097:Repressor Proteins / Q000235:genetics; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D014157:Transcription Factors / Q000235:genetics; D014178:Translocation, Genetic; D020931:ran GTP-Binding Protein / Q000235:genetics; D000097764:ETS Translocation Variant 6 Protein
D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D011495:Histone-Lysine N-Methyltransferase; D051788:Myeloid-Lymphoid Leukemia Protein; D015513:Oncogene Proteins; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D051562:Proto-Oncogene Proteins c-bcr; D050783:Proto-Oncogene Proteins c-ets; D012097:Repressor Proteins; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D014157:Transcription Factors; D020931:ran GTP-Binding Protein; D000097764:ETS Translocation Variant 6 Protein; C497422:KMT2A protein, human; C417257:RanGTP-binding protein 17; C118089:TFPT protein, human; C067143:TAL1 protein, human; C496613:BCR protein, human