title: Pre-clinical evaluation of probes to detect t(8;21) AML minimal residual disease by fluorescence in situ hybridization.
abstract: The 8;21 translocation in acute myeloid leukemia (AML) results in a consistent fusion transcript, AML1/ETO. Long-term clinical remission occurs in some patients despite incomplete eradication of AML1/ETO as demonstrated by RT-PCR, thus limiting the usefulness of this assay. An important future goal will be to determine if there is a level of minimal residual disease (MRD) in patients below which relapse is unlikely. For the detection of MRD, we have developed reagents for fluorescence in situ hybridization (FISH) that identify both derivative 8 and 21 chromosomes with a high analytical sensitivity. In t(8;21) AML cells, two fused signals were detected in addition to the normal 8 and 21 alleles. The sensitivity and specificity of this probe mixture were analyzed in cell lines and patient bone marrows. One and two randomly juxtaposed signals were observed in 2.4 and 0.04% of normal cells, respectively. However, these were easily differentiated from t(8;21) cells by the absence of signals from the normal alleles. Using as criteria the presence of two fused signals plus the normal alleles, we observed no false positives among 5,000 normal cells. The probe correctly identified 20/20 patients with t(8;21) AML and 10/10 non-t(8;21) patients. In cell dilution experiments, the analytical sensitivity of this reagent was equal to that of the X chromosome and Y chromosome alpha-satellite probes. These optimized probes should facilitate the quantitative assessment and study of MRD in t(8;21) AML.
authors.affiliation: University of Colorado Health Sciences Center, Medical Oncology Division, Denver, USA.
authors.forename: G A
authors.initials: GA
authors.lastname: Paskulin
authors.forename: G
authors.initials: G
authors.lastname: Philips
authors.forename: R
authors.initials: R
authors.lastname: Morgan
authors.forename: A
authors.initials: A
authors.lastname: Sandberg
authors.forename: K
authors.initials: K
authors.lastname: Richkind
authors.forename: C
authors.initials: C
authors.lastname: Borovik
authors.forename: L
authors.initials: L
authors.lastname: McGavran
authors.forename: N
authors.initials: N
authors.lastname: Rabinovich
authors.forename: J
authors.initials: J
authors.lastname: Dietz-Band
authors.forename: P
authors.initials: P
authors.lastname: Erickson
authors.forename: H
authors.initials: H
authors.lastname: Drabkin
authors.forename: M
authors.initials: M
authors.lastname: Varella-Garcia
mesh_terms: D001854:Bone Marrow Cells / Q000473:pathology; D002874:Chromosome Mapping; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics*; D002898:Chromosomes, Human, Pair 8 / Q000235:genetics*; D050676:Core Binding Factor Alpha 2 Subunit; D015342:DNA Probes; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis / Q000235:genetics*; D018365:Neoplasm, Residual / Q000175:diagnosis / Q000235:genetics*; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics; D014178:Translocation, Genetic
chemical_list: C107844:AML1-ETO fusion protein, human; D050676:Core Binding Factor Alpha 2 Subunit; D015342:DNA Probes; D005456:Fluorescent Dyes; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors