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Paper Information

Journal:   INTERNATIONAL JOURNAL OF HEMATOLOGY-ONCOLOGY AND STEM CELL RESEARCH (IJHOSCR)   JULY 2016 , Volume 10 , Number 3; Page(s) 147 To 152.
 
Paper: 

EVALUATION OF MINIMAL RESIDUAL DISEASE IN ACUTE MYELOID LEUKEMIA WITH NPM1 MARKER

 
 
Author(s):  ALIZAD GHANDFOROUSH NASRIN, CHAHARDOULI BAHRAM, ROSTAMI SHAHRBANO, GHADIMI HABIBEH, GHASEMI ALI, ALIMOGHADDAM KAMRAN, GHAVAMZADEH ARDESHIR, NADALI FATEMEH*
 
* DEPARTMENT OF HEMATOLOGY, SCHOOL OF ALLIED MEDICAL SCIENCES, TEHRAN UNIVERSITY OF MEDICAL SCIENCES, TEHRAN, IRAN
 
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.

 
Keyword(s): ACUTE MYELOID LEUKEMIA, MRD, NPM1 MUTATION, Q-RT-PCR
 
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