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Analysis of Chromosomal Translocations

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Medical Biomethods Handbook

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Abstract

Chromosomal translocations were the first target for the specific detection of residual tumor cells in bone marrow and peripheral blood. Some types of leukemia are regularly or generally associated with translocations. In chronic myelogenous leukemia (CML) and a proportion of patients with acute lymphoblastic leukemia (ALL), the t(9;22)(q34;q11) translocation leads to the fusion of the ABL gene with part of the BCR gene. Locating the primers such that the polymerase chain reaction (PCR) product spans the fusion point makes it possible to amplify sequences of the hybrid gene specifically (see Chapter 6). Because the breakpoints are better defined at the RNA level than at the DNA level, the primary target is the RNA, which is transcribed into cDNA by reverse transcription. (RT-PCR). In this way, a single leukemic cell can be detected from among 106 normal bone marrow or peripheral blood cells. Comparable approaches have been applied to other fusions such as the PML-RARα fusion gene in acute promyelocytic leukemia (APL). The presence of a few cells with phenotypic or genotypic features of leukemia cells after therapeutic regimens such as stem cell transplantation is know as “minimal residual disease” (MRD).

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References

  1. Wagener, C. (1997) Molecular diagnostics. J. Mol. Med. 75, 728–744.

    Article  PubMed  CAS  Google Scholar 

  2. Rowley, J. D. (1998) The critical role of chromosome translocations in human leukemias. Annu. Rev. Genet. 32, 495–519.

    Article  PubMed  CAS  Google Scholar 

  3. Nowell, P. C. (2002) Progress with chronic myelogenous leukemia: a personal perspective over four decades. Annu. Rev. Med. 53, 1–13.

    Article  PubMed  CAS  Google Scholar 

  4. Bloomfield, C. D., Lawrence, D., Byrd, J. C., et al. (1998) Frequency of prolonged remission duration after high-dose cytarabine intensification in acute myeloid leukemia varies by cytogenetic subtype. Cancer Res. 58, 4173–4179.

    PubMed  CAS  Google Scholar 

  5. Cross, N. C. and Reiter, A. (2002) Tyrosine kinase fusion genes in chronic myeloproliferative diseases. Leukemia 16, 1207–1212.

    Article  PubMed  CAS  Google Scholar 

  6. Nowell, P. C. and Hungerford, D. A. (1960) A minute chromosome in human chronic granulocytic leukemia. Science 132, 1497–1501

    Google Scholar 

  7. Rowley, J. D. (1973) A new consistent chromosome abnormality in chronic myelogenous leukemia detected by quinacrine fluorescence and Giemsa staining. Nature 243, 290–293.

    Article  PubMed  CAS  Google Scholar 

  8. Groffen, J., Stephenson, J. R., Heisterkamp, N., de Klein, A., Bartram, C. R., and Grosveld, G. (1984) Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22. Cell 36, 93–99.

    Article  PubMed  CAS  Google Scholar 

  9. Stam, K., Heisterkamp, N., Grosveld, G., et al. (1985) Evidence of a new chimeric bcrc-abl mRNA in patients with chronic myelocytic leukemia and the Philadelphia chromosome. N. Engl. J. Med. 313, 1429–1433.

    Article  PubMed  CAS  Google Scholar 

  10. Hook, E. B. (1977) Exclusion of chromosomal mosaicism: tables of 90‰, 95‰, and 99‰ confidence limits and comments on use. Am. J. Hum. Genet. 29, 94–97.

    PubMed  CAS  Google Scholar 

  11. Kantarjian, H. M., Dixon, D., Keating, M. J., et al. (1988) Characteristics of accelerated disease in chronic myelogenous leukemia. Cancer 61, 1441–1446.

    Article  PubMed  CAS  Google Scholar 

  12. Hochhaus A, Weisser A, La Rosée P, et al. (2000) Detection and quantification of residual disease in chronic myelogenous leukemia. Leukemia 14, 998–1005.

    Article  PubMed  CAS  Google Scholar 

  13. Tkachuk, D. C., Westbrook, C. A., Andreeff, M., et al. (1990) Detection of BCR-ABL fusion in chronic myelogeneous leukemia by in situ hybridization. Science 250, 559–562.

    Article  PubMed  CAS  Google Scholar 

  14. Weber-Matthiesen, K., Winkemann, M., Müller-Hermelink, A., Schlegelberger, B., and Grote, W. (1992) Simultaneous fluorescence immunophenotyping and interphase cytogenetics: a contribution to characterization of tumor cells. J. Histochem. Cytochem. 40, 171–175.

    PubMed  CAS  Google Scholar 

  15. Chase, A., Grand, F., Zhang, J. G., Blackett, N., Goldman, J., and Gordon, M. (1997) Factors influencing the false positive and negative rates of BCR-ABL fluorescence in-situ hybridization. Genes Chromosomes Cancer 18, 246–253.

    Article  PubMed  CAS  Google Scholar 

  16. Sinclair, P. B., Green, A. R., Grace, C., and Nacheva, E. P. (1997) Improved sensitivity of BCRABL detection: a triple-probe three-color fluorescence in situ hybridization system. Blood 90, 1395–1402.

    PubMed  CAS  Google Scholar 

  17. Dewald, G. W., Wyatt, W. A., Juneau, A. L., et al. (1998) Highly sensitive fluorescence in situ hybridization method to detect double BCR-ABL fusion and monitor response to therapy in chronic myeloid leukemia. Blood 91, 3357–3365.

    PubMed  CAS  Google Scholar 

  18. Seong, D. C., Kantarjian, H. M., Ro, J. Y., et al. (1995) Hypermetaphase fluorescence in situ hybridization for quantitative monitoring of Philadelphia chromosome-positive cells in patients with chronic myelogenous leukemia during treatment. Blood 86, 2343–2349.

    PubMed  CAS  Google Scholar 

  19. Reiter, A., Skladny, H., Hochhaus, A., et al. (1997) Molecular response of CML patients treated with interferon-monitored by quantitative Southern blot analysis. Br. J. Haematol. 97, 86–93.

    Article  PubMed  CAS  Google Scholar 

  20. Verschraegen, C. F., Talpaz, M., Hirsch Ginsberg, C. F., et al. (1995) Quantification of the breakpoint cluster region rearrangement for clinical monitoring in Philadelphia chromosome-positive chronic myeloid leukemia. Blood 85, 2705–2710.

    PubMed  CAS  Google Scholar 

  21. Guo, J. Q., Lian, J. Y., Xian, Y. M., et al. (1994) BCR-ABL protein expression in peripheral blood cells of chronic myelogenous leukemia patients undergoing therapy. Blood 83, 3629–3637.

    PubMed  CAS  Google Scholar 

  22. Morgan, G. J., Hughes, T., Janssen, J. W., et al. (1989) Polymerase chain reaction for detection of residual leukemia. Lancet 1, 928–929.

    Article  PubMed  CAS  Google Scholar 

  23. Cross, N. C. P. (1997) Assessing residual leukemia. Baillieres Clin. Haematol. 10, 389–403.

    Article  PubMed  CAS  Google Scholar 

  24. Cross, N. C. P., Melo, J. V., Feng, L., and Goldman, J. M. (1994) An optimized multiplex polymerase chain reaction (PCR) for detection of BCR-ABL fusion mRNAs in haematological disorders. Leukemia 8, 186–189.

    PubMed  CAS  Google Scholar 

  25. Morley, A. (1998) Quantifying leukemia. N. Engl. J. Med. 339, 627–629.

    Article  PubMed  CAS  Google Scholar 

  26. van Dongen, J. J. M., MacIntyre, E. A., Gabert, J. A., et al. (1999) Standardized RT-PCR analysis of fusion gene transcripts from chromosome aberrations in acute leukemia for detection of minimal residual disease. Leukemia 12, 1901–1928.

    Article  Google Scholar 

  27. Lin, F., van Rhee, F., Goldman, J. M., and Cross, N. C. P. (1996) Kinetics of increasing BCR-ABL transcript numbers in chronic myeloid leukemia patients who relapse after bone marrow transplantation. Blood 87, 4473–4478.

    PubMed  CAS  Google Scholar 

  28. Hochhaus, A., Reiter, A., Sauβele, S., et al., for the German CML Study Group and the UK MRC CML Study Group (2000) Molecular heterogeneity in complete cytogenetic responders after interferontherapy for chronic myelogenous leukemia: low levels of minimal residual disease are associated with continuing remission. Blood 95, 62–66.

    PubMed  CAS  Google Scholar 

  29. Biernaux, C., Loos, M., Sels, A., Huez, G., and Stryckmans, P. (1995) Detection of major bcr-abl gene expression at a very low level in blood cells of some healthy individuals. Blood 88, 3118–3122.

    Google Scholar 

  30. Cross, N. C. P., Feng, L., Chase, A., Bungey, J., Hughes, T. P., and Goldman, J. M. (1993) Competitive polymerase chain reaction to estimate the number of BCR-ABL transcripts in chronic myeloid leukemia patients after bone marrow transplantation. Blood 82, 1929–1936.

    PubMed  CAS  Google Scholar 

  31. Lin, F., Kirkland, M. A., van Rhee, F., et al. (1996) Molecular analysis of transient cytogenetic relapse after allogeneic bone marrow transplantation for chronic myeloid leukemia. Bone Marrow Transplant. 18, 1147–1152.

    PubMed  CAS  Google Scholar 

  32. van Rhee, F., Lin, F., Cullis, J. O., et al. (1994) Relapse of chronic myeloid leukemia after allogeneic bone marrow transplant: the case for giving donor leukocyte transfusions before the onset of hematologic relapse. Blood 83, 3377–3383.

    PubMed  Google Scholar 

  33. Corsetti, M. T., Lerma, E., Dejana, A., et al. (1999) Quantitative competitive reverse transcriptasepolymerase chain reaction for BCR-ABL on Philadelphia-negative leukaphereses allows the selection of low-contaminated peripheral blood progenitor cells for autografting in chronic myelogenous leukemia. Leukemia 13, 999–1008.

    Article  PubMed  CAS  Google Scholar 

  34. Preudhomme, C., Révillion, F., Merlat, A., et al. (1999) Detection of BCR-ABL transcripts in chronic myeloid leukemia (CML) using a’ real time’ quantitative RT-PCR assay. Leukemia 13, 957–964.

    Article  PubMed  CAS  Google Scholar 

  35. Wittwer, C. T., Herrmann, M. G., Moss, A. A., and Rasmussen, R. P. (1997) Continuous fluorescence monitoring of rapid cycle DNA amplification. Biotechniques 22, 130–138.

    PubMed  CAS  Google Scholar 

  36. Emig, M., Saussele, S., Wittor, H., et al. (1999) Accurate and rapid analysis of residual disease in patients with CML using specific fluorescent hybridization probes for real time quantitative RTPCR. Leukemia 13, 1825–1832.

    Article  PubMed  CAS  Google Scholar 

  37. Hughes, T. P., Kaeda, J., Branford, S., for the International Randomized Study of Interferon versus STI571 (IRIS) Study Group. (2003) Frequency of major molecular responses to imatinib or interferon alfa plus cytarabine in newly diagnosed patients with chronic myeloid leukemia. N. Engl. J. Med. 349, 1421–1430.

    Article  Google Scholar 

  38. Müller, M. C., Gattermann, N., Lahaye, T., et al. (2003) Dynamics of BCR-ABL mRNA expression in first line therapy of chronic myelogenous leukemia patients with imatinib or interferon a/ara-C. Leukemia 17, 2392–2400.

    Article  PubMed  Google Scholar 

  39. Schoch, C., Schnittger, S., Bursch, S., et al. (2002) Comparison of chromosome banding analysis, interphase-and hypermetaphase-FISH, qualitative and quantitative PCR for diagnosis and for followup in chronic myeloid leukemia: A study of 350 cases. Leukemia 16, 53–59.

    Article  PubMed  CAS  Google Scholar 

  40. van der Velden, V. H. J., Hochhaus, A., Cazzaniga, G., Szczepanski, T., Gabert, J., and van Dongen, J. J. M. (2003) Detection of minimal residual disease in hematologic malignancies by real-time quantitative PCR: principles, approaches, and laboratory aspects. Leukemia 17, 1013–1034.

    Article  PubMed  Google Scholar 

  41. Gabert, J., Beillard, E., van der Velden, V. H. J., et al. (2003) Standardization and quality control studies of “real-time” quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) of fusion gene transcripts for residual disease detection in leukemia-a Europe Against Cancer Program. Leukemia 17, 2318–2357.

    Article  PubMed  CAS  Google Scholar 

  42. Beillard, E., Pallisgaard, N., van der Velden, V. H. J., et al. (2003) Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using “real-time” quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR)-an Europe Against Cancer Program. Leukemia 17, 2474–2486.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Medizinische Klinik, Fakultät für Klinische Medizin Mannheim der Universität Heidelberg, Mannheim, Germany

    Andreas Hochhaus

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  1. Andreas Hochhaus
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Editors and Affiliations

  1. University of Hertfordshire, Hatfield, UK

    John M. Walker  & Ralph Rapley  & 

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© 2005 Humana Press Inc., Totowa, NJ

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Hochhaus, A. (2005). Analysis of Chromosomal Translocations. In: Walker, J.M., Rapley, R. (eds) Medical Biomethods Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-870-6:471

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  • DOI: https://doi.org/10.1385/1-59259-870-6:471

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-288-9

  • Online ISBN: 978-1-59259-870-0

  • eBook Packages: Springer Protocols

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