Fine structure of translocation breakpoints within the major breakpoint region in BCR-ABL1-positive leukemias

Abstract The chromosomal translocation t(9;22)(q34;q22), with expression of the BCR-ABL1 fusion gene is the cytogenetic and molecular hallmark of chronic myeloid leukemia (CML) and a subset of acute lymphoblastic leukemia (ALL). Basically two types of BCR-ABL1 chimeric mRNA transcripts have been observed: (1) e13a2/e14a2 transcripts in CML and ALL, resulting from chromosomal breaks in the major breakpoint cluster region (M-bcr) of the BCR gene and (2) e1a2 transcripts in ALL resulting from breaks in the minor breakpoint cluster region (m-bcr) of the BCR gene. To gain a better understanding of this molecular alteration, we developed a long-distance inverse PCR (LDI PCR) method for M-bcr breakpoint identification in BCR-ABL1 -positive cases and were thus able to identify the chromosomal breakpoints within the M-bcr in 62 BCR-ABL1 -positive samples. The corresponding reciprocal breakpoints were identified and molecularly characterized in 45 of these cases. In 2 samples, the breaks were located 5′ to the ABL1 locus and in one case, the der(9) break was identified on 9q34.13 several hundred kB 3′ telomeric to ABL1 . The analysis of breaks revealed no significant clustering and no association with repetitive elements (Alu, L1, L2) or recombination signal sequence sites. The established LDI PCR permits fast, relatively easy and unbiased identification of breakpoints in the M-bcr region of BCR and also enables the molecular analysis of more complex translocations with breakpoints outside the ABL1 gene locus or other BCR fusion genes.