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

  • Nasrin Alizad Ghandforoush MSc, Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
  • Bahram Chahardouli Assistant Professor, Hematology-Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Shahrbano Rostami Assistant Professor, Hematology-Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Habibeh Ghadimi MSc Student, Hematology-Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Ali Ghasemi PhD Student of Hematology, Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
  • Kamran Alimoghaddam Professor, Hematology-Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Ardeshir Ghavamzadeh Professor, Hematology-Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran
  • Fatemeh Nadali Mail Associate Professor, Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
Keywords:
Acute myeloid leukemia, MRD, NPM1 mutation, Q-RT-PCR

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. Eleven patients were analyzed using established method. Serial PB and/or BM samples (n=71) were taken in 1-3 months intervals (mean 1.5-month intervals) and median follow-up duration after chemotherapy was 11 months (5-28.5 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 757 with median of 383.5 in samples at diagnosis. The median NPMmut transcript level log reduction was 3 logs. Relapse occurred in 54.5% of patients (n=6), 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=2), 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: Despite the limitations of this study in terms of sample size and duration of follow-up, it showed the accuracy of set up for detection of mutation and this marker has worth for following-up at different stages of disease. Because of high frequency, stability, specificity to abnormal clone and high sensitivity, NPM1 is a suitable marker for monitoring of NPMc+ AML patients.

 

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Published
2016-06-28
How to Cite
1.
Alizad Ghandforoush N, Chahardouli B, Rostami S, Ghadimi H, Ghasemi A, Alimoghaddam K, Ghavamzadeh A, Nadali F. Evaluation of Minimal Residual Disease in Acute Myeloid Leukemia with NPM1 Marker. Int J Hematol Oncol Stem Cell Res. 10(3):147-152.
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Articles