Abstract
We sought to understand the genesis of the t(9;22) by characterizing genomic breakpoints in chronic myeloid leukemia (CML) and BCR–ABL-positive acute lymphoblastic leukemia (ALL). BCR–ABL breakpoints were identified in p190 ALL (n=25), p210 ALL (n=25) and p210 CML (n=32); reciprocal breakpoints were identified in 54 cases. No evidence for significant clustering and no association with sequence motifs was found except for a breakpoint deficit in repeat regions within BCR for p210 cases. Comparison of reciprocal breakpoints, however, showed differences in the patterns of deletion/insertions between p190 and p210. To explore the possibility that recombinase-activating gene (RAG) activity might be involved in ALL, we performed extra-chromosomal recombination assays for cases with breakpoints close to potential cryptic recombination signal sequence (cRSS) sites. Of 13 ALL cases tested, 1/10 with p190 and 1/3 with p210 precisely recapitulated the forward BCR–ABL breakpoint and 1/10 with p190 precisely recapitulated the reciprocal breakpoint. In contrast, neither of the p210 CMLs tested showed functional cRSSs. Thus, although the t(9;22) does not arise from aberrant variable (V), joining (J) and diversity (D) (V(D)J) recombination, our data suggest that in a subset of ALL cases RAG might create one of the initiating double-strand breaks.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Deininger MW, Bose S, Gora-Tybor J, Yan XH, Goldman JM, Melo JV . Selective induction of leukemia-associated fusion genes by high-dose ionizing radiation. Cancer Res 1998; 58: 421–425.
Stanulla M, Wang J, Chervinsky DS, Thandla S, Aplan PD . DNA cleavage within the MLL breakpoint cluster region is a specific event which occurs as part of higher-order chromatin fragmentation during the initial stages of apoptosis. MolCell Biol 1997; 17: 4070–4079.
Neves H, Ramos C, da Silva MG, Parreira A, Parreira L . The nuclear topography of ABL, BCR, PML, and RARalpha genes: evidence for gene proximity in specific phases of the cell cycle and stages of hematopoietic differentiation. Blood 1999; 93: 1197–1207.
Reiter A, Saussele S, Grimwade D, Wiemels JL, Segal MR, Lafage-Pochitaloff M et al. Genomic anatomy of the specific reciprocal translocation t(15;17) in acute promyelocytic leukemia. Genes Chromosomes Cancer 2003; 36: 175–188.
Mattarucchi E, Guerini V, Rambaldi A, Campiotti L, Venco A, Pasquali F et al. Microhomologies and interspersed repeat elements at genomic breakpoints in chronic myeloid leukemia. Genes Chromosomes Cancer 2008; 47: 625–632.
Melo JV . BCR-ABL gene variants. Baillieres Clin Haematol 1997; 10: 203–222.
van der Feltz MJ, Shivji MK, Allen PB, Heisterkamp N, Groffen J, Wiedemann LM . Nucleotide sequence of both reciprocal translocation junction regions in a patient with Ph positive acute lymphoblastic leukaemia, with a breakpoint within the first intron of the BCR gene. Nucleic Acids Res 1989; 17: 1–10.
Chen SJ, Chen Z, Font MP, d’Auriol L, Larsen CJ, Berger R . Structural alterations of the BCR and ABL genes in Ph1 positive acute leukemias with rearrangements in the BCR gene first intron: further evidence implicating Alu sequences in the chromosome translocation. Nucleic Acids Res 1989; 17: 7631–7642.
Jeffs AR, Benjes SM, Smith TL, Sowerby SJ, Morris CM . The BCR gene recombines preferentially with Alu elements in complex BCR-ABL translocations of chronic myeloid leukaemia. Hum Mol Genet 1998; 7: 767–776.
Chen SJ, Chen Z, d’Auriol L, Le Coniat M, Grausz D, Berger R . Ph1+bcr- acute leukemias: implication of Alu sequences in a chromosomal translocation occurring in the new cluster region within the BCR gene. Oncogene 1989; 4: 195–202.
Robbiani DF, Bothmer A, Callen E, Reina-San-Martin B, Dorsett Y, Difilippantonio S et al. AID is required for the chromosomal breaks in c-myc that lead to c-myc/IgH translocations. Cell 2008; 135: 1028–1038.
Tsai AG, Lu H, Raghavan SC, Muschen M, Hsieh CL, Lieber MR . Human chromosomal translocations at CpG sites and a theoretical basis for their lineage and stage specificity. Cell 2008; 135: 1130–1142.
Marculescu R, Vanura K, Montpellier B, Roulland S, Le T, Navarro JM et al. Recombinase, chromosomal translocations and lymphoid neoplasia: targeting mistakes and repair failures. DNA Repair (Amst) 2006; 5: 1246–1258.
Roth DB . Restraining the V(D)J recombinase. Nat Rev Immunol 2003; 3: 656–666.
Brandt VL, Roth DB . Recent insights into the formation of RAG-induced chromosomal translocations. Adv Exp Med Biol 2009; 650: 32–45.
Aplan PD, Lombardi DP, Ginsberg AM, Cossman J, Bertness VL, Kirsch IR . Disruption of the human SCL locus by ‘illegitimate’ V-(D)-J recombinase activity. Science 1990; 250: 1426–1429.
Raghavan SC, Swanson PC, Ma Y, Lieber MR . Double-strand break formation by the RAG complex at the bcl-2 major breakpoint region and at other non-B DNA structures in vitro. Mol Cell Biol 2005; 25: 5904–5919.
Fialkow PJ, Denman AM, Jacobson RJ, Lowenthal MN . Chronic myelocytic leukemia. Origin of some lymphocytes from leukemic stem cells. J Clin Invest 1978; 62: 815–823.
Cobaleda C, Gutierrez-Cianca N, Perez-Losada J, Flores T, Garcia-Sanz R, Gonzalez M et al. A primitive hematopoietic cell is the target for the leukemic transformation in human philadelphia-positive acute lymphoblastic leukemia. Blood 2000; 95: 1007–1013.
Schenk TM, Keyhani A, Bottcher S, Kliche KO, Goodacre A, Guo JQ et al. Multilineage involvement of Philadelphia chromosome positive acute lymphoblastic leukemia. Leukemia 1998; 12: 666–674.
Zaliova M, Fronkova E, Krejcikova K, Muzikova K, Mejstrikova E, Stary J et al. Quantification of fusion transcript reveals a subgroup with distinct biological properties and predicts relapse in BCR/ABL-positive ALL: implications for residual disease monitoring. Leukemia 2009; 23: 944–951.
Bacher U, Haferlach T, Hiddemann W, Schnittger S, Kern W, Schoch C . Additional clonal abnormalities in Philadelphia-positive ALL and CML demonstrate a different cytogenetic pattern at diagnosis and follow different pathways at progression. Cancer Genet Cytogenet 2005; 157: 53–61.
Turhan AG, Eaves CJ, Kalousek DK, Eaves AC, Humphries RK . Molecular analysis of clonality and bcr rearrangements in Philadelphia chromosome-positive acute lymphoblastic leukemia. Blood 1988; 71: 1495–1498.
Hotfilder M, Rottgers S, Rosemann A, Schrauder A, Schrappe M, Pieters R et al. Leukemic stem cells in childhood high-risk ALL/t(9;22) and t(4;11) are present in primitive lymphoid-restricted CD34+. Cancer Res 2005; 65: 1442–1449.
Castor A, Nilsson L, Astrand-Grundstrom I, Buitenhuis M, Ramirez C, Anderson K et al. Distinct patterns of hematopoietic stem cell involvement in acute lymphoblastic leukemia. Nat Med 2005; 11: 630–637.
Melo JV, Myint H, Galton DA, Goldman JM . P190BCR-ABL chronic myeloid leukaemia: the missing link with chronic myelomonocytic leukaemia? Leukemia 1994; 8: 208–211.
Li S, Ilaria Jr RL, Million RP, Daley GQ, Van Etten RA . The P190, P210, and P230 forms of the BCR/ABL oncogene induce a similar chronic myeloid leukemia-like syndrome in mice but have different lymphoid leukemogenic activity. J Exp Med 1999; 189: 1399–1412.
Cross NCP, Melo JV, Feng L, Goldman JM . An optimized multiplex polymerase chain reaction (PCR) for detection of BCR-ABL fusion mRNAs in haematological disorders. Leukemia 1994; 8: 186–189.
Baxter EJ, Kulkarni S, Vizmanos JL, Jaju R, Martinelli G, Testoni N et al. Novel translocations that disrupt the platelet-derived growth factor receptor beta (PDGFRB) gene in BCR-ABL-negative chronic myeloproliferative disorders. Br J Haematol 2003; 120: 251–256.
Kreil S, Pfirrmann M, Haferlach C, Waghorn K, Chase A, Hehlmann R et al. Heterogeneous prognostic impact of derivative chromosome 9 deletions in chronic myelogenous leukemia. Blood 2007; 110: 1283–1290.
Segal MR, Wiemels JL . Clustering of translocation breakpoints. Journal of the American Statistical Association 2002; 97: 66–76.
Gellert M . V(D)J recombination: RAG proteins, repair factors, and regulation. Annu Rev Biochem 2002; 71: 101–132.
Wiemels JL, Leonard BC, Wang Y, Segal MR, Hunger SP, Smith MT et al. Site-specific translocation and evidence of postnatal origin of the t(1;19) E2A-PBX1 fusion in childhood acute lymphoblastic leukemia. Proc Natl Acad Sci USA 2002; 99: 15101–15106.
Marculescu R, Le T, Simon P, Jaeger U, Nadel B . V(D)J-mediated translocations in lymphoid neoplasms: a functional assessment of genomic instability by cryptic sites. J Exp Med 2002; 195: 85–98.
Weinstock DM, Jasin M . Alternative pathways for the repair of RAG-induced DNA breaks. Mol Cell Biol 2006; 26: 131–139.
Weinstock DM, Brunet E, Jasin M . Formation of NHEJ-derived reciprocal chromosomal translocations does not require Ku70. Nat Cell Biol 2007; 9: 978–981.
Nowell PC, Hungerford DA . Chromosome studies on normal and leukemic human leukocytes. J NatlCancer Inst 1960; 25: 85–109.
Reichel M, Gillert E, Nilson I, Siegler G, Greil J, Fey GH et al. Fine structure of translocation breakpoints in leukemic blasts with chromosomal translocation t(4;11): the DNA damage-repair model of translocation. Oncogene 1998; 17: 3035–3044.
Zhang JG, Goldman JM, Cross NCP . Characterization of genomic BCR-ABL breakpoints in chronic myeloid leukaemia by PCR. BrJ Haematol 1995; 90: 138–146.
Tycko B, Sklar J . Chromosomal translocations in lymphoid neoplasia: a reappraisal of the recombinase model. Cancer Cells 1990; 2: 1–8.
Raghavan SC, Swanson PC, Wu X, Hsieh CL, Lieber MR . A non-B-DNA structure at the Bcl-2 major breakpoint region is cleaved by the RAG complex. Nature 2004; 428: 88–93.
Pasqualucci L, Bhagat G, Jankovic M, Compagno M, Smith P, Muramatsu M et al. AID is required for germinal center-derived lymphomagenesis. Nat Genet 2008; 40: 108–112.
Chen SJ, Chen Z, Hillion J, Grausz D, Loiseau P, Flandrin G et al. Ph1-positive, bcr-negative acute leukemias: clustering of breakpoints on chromosome 22 in the 3′ end of the BCR gene first intron. Blood 1989; 73: 1312–1315.
Chen SJ, Chen Z, Font MP, d’Auriol L, Larsen CJ, Berger R . Structural alterations of the BCR and ABL genes in Ph1 positive acute leukemias with rearrangements in the BCR gene first intron: further evidence implicating Alu sequences in the chromosome translocation. Nucleic Acids Res 1989; 17: 7631–7642.
Melo JV, Gordon DE, Tuszynski A, Dhut S, Young BD, Goldman JM . Expression of the ABL-BCR fusion gene in Philadelphia-positive acute lymphoblastic leukemia. Blood 1993; 81: 2488–2491.
Specchia G, Albano F, Anelli L, Storlazzi CT, Zagaria A, Mancini M et al. Deletions on der(9) chromosome in adult Ph-positive acute lymphoblastic leukemia occur with a frequency similar to that observed in chronic myeloid leukemia. Leukemia 2003; 17: 528–531.
Huntly BJ, Reid AG, Bench AJ, Campbell LJ, Telford N, Shepherd P et al. Deletions of the derivative chromosome 9 occur at the time of the Philadelphia translocation and provide a powerful and independent prognostic indicator in chronic myeloid leukemia. Blood 2001; 98: 1732–1738.
Acknowledgements
This work was funded by Leukaemia and Lymphoma Research (UK); JS was supported by a Gordon Piller PhD Studentship.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Author contributions
The study was designed by JS, NCPC and FHG. Laboratory analysis was performed by JS, MAS-S, SK, CH-C, DW. Data analysis was performed by JS, RFY, JW, BN. Samples and data were provided by MJC, OO, FP, MAS-S, JVM. The initial paper was written by JS, NCPC and FHG; all authors contributed to the final version.
Supplementary Information accompanies the paper on the Leukemia website
Supplementary information
Rights and permissions
About this article
Cite this article
Score, J., Calasanz, M., Ottman, O. et al. Analysis of genomic breakpoints in p190 and p210 BCR–ABL indicate distinct mechanisms of formation. Leukemia 24, 1742–1750 (2010). https://doi.org/10.1038/leu.2010.174
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/leu.2010.174
Keywords
This article is cited by
-
The importance of personalized medicine in chronic myeloid leukemia management: a narrative review
Egyptian Journal of Medical Human Genetics (2023)
-
Characterization of p190-Bcr-Abl chronic myeloid leukemia reveals specific signaling pathways and therapeutic targets
Leukemia (2021)
-
Treatment-free remission in patients with chronic myeloid leukaemia
Nature Reviews Clinical Oncology (2020)
-
Is cancer latency an outdated concept? Lessons from chronic myeloid leukemia
Leukemia (2020)
-
Aberrant RAG-mediated recombination contributes to multiple structural rearrangements in lymphoid blast crisis of chronic myeloid leukemia
Leukemia (2020)