Evaluation of Minimal Residual Disease in Acute Myeloid Leukemia with NPM1 Marker

ABSTRACT Background: Minimal residual disease (MRD) tests provide early identification of hematologic relapse and timely management of acute myeloid leukemia (AML) patients. Approximately, 50% of AML patients do not have clonal chromosomal aberrations and categorize as a cytogenetically normal acute myeloid leukemia (CN-AML). About 60% of adult CN-AML has a mutation in exon 12 of NPM1 gene. This mutation is specific for malignant clone and potentially is a good marker of MRD. In this retrospective study, we set up a quantitative test for quantifying NPM1 type A mutation and AML patients carrying this mutation at the time of diagnosis, were followed-up.Materials and Methods: We prepared plasmids containing a cDNA fragment of NPM1 and ABL genes by PCR cloning. The plasmids were used to construct standard curves. twelve patients were analyzed using established method. Serial PB and/or BM samples (n=101) were taken in 1-3 months intervals (mean 1.5-month intervals) and median follow-up duration after chemotherapy was 13 months (5-30.5 months). Results: In this study, we developed RNA-based RQ-PCR to quantitation of NPM1 mutation A with sensitivities of 10(-5). The percent of NPMmut/ABL level showed a range between 132 and 4388 with median of 907 in samples at diagnosis. The median NPMmut transcript level log reduction was 3.3 logs. Relapse occurred in 58.33% of patients (n=7), all cases at diagnosis demonstrated the same mutation at relapse. In patients who experienced relapse, log reduction levels of NPM1 mRNA transcript after therapy were 4 (n=3), 3 (n=2) and 1 log (n=2). Totally, NPMmut level showed less than 5 log reduction in all of them, whereas this reduction was 5-6 logs in other patients. Conclusion: Because of high frequency, stability, specificity to abnormal clone and high sensitivity, NPM1 is a suitable marker for monitoring of NPMc+ AML patients.

Evaluation of Minimal Residual Disease in Acute Myeloid Leukemia with NPM1 Marker Keywords:

NPM1 , MRD , AML