Development of a dual-color fluorescence in situ hybridization probe set on chromosome 6q to improve cytogenetic diagnosis of lymphoid malignancies

Butler, Melody; Ronnenburg, Michael J.; Ament, Candice; Meekins, Joann; Ning, Yi

doi:10.1016/j.cancergencyto.2004.06.008

Cited by 3 publications

(2 citation statements)

References 15 publications

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“…These two clones from the human BAC library were obtained from Children's Hospital Oakland Research Institute (Oakland, CA, USA). DNA isolated from these clones were labeled with fluorochromes following the protocol described in our previous studies 6 and validated as a FISH probe set on normal controls. This probe set was then used to confirm the 8;16 translocation in both patients.…”

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confidence: 99%

Detection of MOZ-CBP fusion in acute myeloid leukemia with 8;16 translocation

et al. 2005

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Chromosome translocations that result in gene fusion and expression of the fusion gene products are frequently observed in acute myeloid leukemia (AML). The recurrent translocation t(8;16)(p11;p13) is a cytogenetic hallmark for the M4/M5 subtype of AML that occurs in both de novo AML and therapy-related AML. This subtype of AML is associated with monocytoid blast morphology, erythrophagocytosis, and a poor prognosis, with a median survival of only 2 months. 1 Molecular studies of the 8;16 translocation revealed a fusion of a histone acetyltransferase MOZ from chromosome 8p11 with a transcriptional adaptor/coactivator CBP from chromosome 16p13. 2,3 Despite accurate mapping of the translocation breakpoints, attempts to amplify the MOZ-CBP and CBP-MOZ transcripts using reverse transcriptase-polymerase chain reaction (RT-PCR) was successful only in a limited number of cases. [2][3][4] Various explanations, such as low expression or high instability, have been proposed for the failure of RT-PCR analysis. Fluorescence in situ hybridization (FISH) is an alternative method for the detection of a specific translocation and for monitoring minimal residual disease. Although a few cosmid clones in the CBP region have been used as FISH probes 3,5 to demonstrate the disruption of the CBP gene, no FISH probe targeting MOZ-CBP fusion is currently available to diagnostic laboratories.Taking advantage of the completed human genome project, we searched the National Center for Biotechnology Information (NCBI) database and identified bacteria-derived artificial chromosome (BAC) clones in the MOZ and CBP regions, respectively. BAC clones with an average insert size of 70-100 kb are ideal material to be used as FISH probes. Using the identified BAC clones, we demonstrated the detection of a MOZ-CBP fusion in two AML patients with an 8;16 translocation. Furthermore, we used this probe set to monitor the disease status during the treatment of a patient who has achieved complete cytogenetic remission for 1.5 years. We also identified a MOZ-CBP transcript by RT-PCR and cDNA sequencing in this patient.Our new FISH probe set was applied on bone marrow specimens from two AML patients. Patient 1 was a 57-year-old female who had breast cancer in 2001 and developed AML in 2003. Cytogenetic analysis detected multiple rearrangements in all of the 20 cells analyzed. Consistent abnormalities included a balanced 8;16 translocation and an unbalanced 1;10 translocation. Three cells had additional rearrangements involving chromosome 12. The karyotype designation was 46,XX,t(8;16) (p11.2;p13.2),der(10)t(1;10)(q21;q26)[17]/46,XX,t(8;16)(p11.2; p13.2),der(10)t(1;10)(q21;q26),add (12) We undertook a database search approach to efficiently identify targeted BAC clones. RP11-108L9 is centromeric and adjacent to the MOZ gene, and RP11-462G12 is telomeric and adjacent to the CBP gene. These two clones from the human BAC library were obtained from Children's Hospital Oakland Research Institute (Oakland, CA, USA). DNA isolated from these clones were labeled with...

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Section: To the Editormentioning

confidence: 99%

Detection of MOZ-CBP fusion in acute myeloid leukemia with 8;16 translocation

et al. 2005

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“…Isolated DNA from this clone was labeled with fluorochromes following the protocol described in our previous studies 5 and validated as a FISH probe on normal controls. This probe was then used to characterize dmins in our patient.…”

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REL-positive double minute chromosomes in follicular lymphoma

et al. 2006

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t(5;10)(q33;q21). 7 A common structural feature of TPM3 and H4/D10S170 proteins is the coiled-coil domain, which contributes with its dimerization/activation moiety to building tyrosine kinase fusion proteins.Given these findings, a double color double fusion FISH assay with clone CTD-2601I11 encompassing the PDGFRB breakpoint, and clone RP11-205M9 for TPM3 (Figure 2) is a valid diagnostic test for discriminating between t(1;5)(q21;q33)/ TPM3/PDGFRB, as described in the present letter, and t(1;5) involving other genes at 1q21-q23 such as PDE4DIP-PDGFRB, which has been reported in a pediatric case of chronic myeloproliferative disorder.

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