Short communicationFeasibility of using the combined MDS-EVI1/EVI1 gene expression as an alternative molecular marker in acute myeloid leukemia: a report of four cases
Introduction
In acute myeloid leukemia (AML), molecular markers are increasingly important for the assessment of prognosis and for post-remission controls to detect minimal residual disease (MRD). These markers include fusion genes [1], [2], [3], [4], gene mutations (e.g., FLT3-LM) [5], [6], and overexpression of oncogenes (e.g., WT1) [7], [8]. The protooncogene EVI1, which is located in the chromosomal region 3q26, is involved in the pathogenesis of AML with 3q26 aberrations [9], [10], [11], [12]. In the majority of reports, EVI1 was not detectable in the normal bone marrow or blood of healthy individuals [13], [14], [15], [16]. One report, however, demonstrated low levels of EVI1 in the bone marrow of healthy volunteers, whereas EVI1 was undetectable in the peripheral blood of the same cohort [17]. On the other hand, there have been reports of AML without 3q26 aberrations expressing EVI1[14], [16], [17], [18]. In addition, it was reported that high levels of EVI1 expression upon diagnosis of AML were associated with a poor survival [17]. Other reports however, have failed to demonstrate a significant influence of initial EVI1 expression on the outcome of AML [19]. This may be due to the presence of two different splice variants, the 3′ EVI1 and the 5′ MDS1/EVI1 variant. Because not all studies performed differential assays for these variants, the effects of EVI1 on prognosis are still conflicting.
Independently of any prognostic relevance of initial expression levels, the aim of the present study was to establish a further follow-up marker for minimal residual disease detection in AML. To increase the percentage of samples that show initial overexpression of this potential MRD marker and thus increase sensitivity, the primer set was designed intentionally to span both the MDS-EVI1 and the EVI1 transcripts. Here, we describe the molecular course of the combined MDS1-EVI1/ EVI1 gene expression levels as assessed by quantitative real-time RT-PCR during anti-leukemic therapy, and we have exemplified this in four patients.
Section snippets
Patients
Bone marrow samples of four patients were analyzed at the Laboratory for Leukemia Diagnostics. All cases were diagnosed as AML according to the French–American–British (FAB) and World Health Organization classification [20], [21], [22]. In total, 33 bone marrow samples from 4 patients were analyzed for this study.
Cytogenetic analysis
Cytogenetic analysis was performed according to standard protocols [23]. Cytogenetic data were classified according to ISCN nomenclature [24]. At least 25 metaphases were analyzed.
Quantitative real-time polymerase chain reaction (PCR)
Results and discussion
Sensitive monitoring of minimal residual disease is of major importance in modern hematology. RT-PCR–based quantification of MRD often relies on quantitative detection of classic fusion transcripts arising from chromosomal translocations. In AML, however, such translocations are available only in a minority of cases [29], [30]. Thus, additional molecular markers for post-remission control are needed. In the present report, we describe the applicability of the combined MDS-EVI1/EVI1 gene as a
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High GATA2 expression is a poor prognostic marker in pediatric acute myeloid leukemia
2012, BloodCitation Excerpt :In 2 of 38 AML patients, GATA2 was sufficiently highly expressed to allow detection of at least a 2-log reduction (ie, sensitivity < 0.01). GATA2 expression was assessed in parallel with additional markers that have been used previously for the detection of residual disease: WT1 expression (n = 38), AML1-ETO fusion gene expression [patients with t(8;21); n = 3], and EVI1 expression (patients with high EVI1 expression, n = 3; Figure 6).6,44,47,48 The expression patterns of GATA2 were comparable to the expression patterns of the AML1-ETO fusion gene, EVI1 and WT1.
High EVI1 levels predict adverse outcome in acute myeloid leukemia: Prevalence of EVI1 overexpression and chromosome 3q26 abnormalities underestimated
2008, BloodCitation Excerpt :A diagnostic assay should be rapid and quantitative and it should take into account each of the different EVI1 splice forms. A multiplex PCR assay, by which the expression levels of each of the different EVI1 splice variants26,27 will be determined separately, is feasible but may be complicated to interpret. It may also be possible to develop primer probe combinations, which recognize all splice forms in one single EVI1-specific Q-PCR.
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