The development of a flow cytometric method to detect the presence of mutated nucleophosmin in Acute Myeloid Leukaemia
Nucleophosmin (NPM1) is an acidic, nucleo-cytoplasmic, shuttling protein with predominant nucleolar localisation that plays multiple roles in cell growth and proliferation. Deletion insertion mutations of NPM1 (NPM1 DIM) seem to disrupt it normal physiologic role as a molecular chaperone, which likely leads to its oncogenic potential.NPM1 if present alone (not associated with FLT3 internal tandem duplications (FLT3-ITD)) is associated with significantly better overall survival and disease free survival in AML and has been entered as a provisional category in the World Health Organisation (2008) classification of Acute Myeloid Leukaemia with recurrent genetic abnormalities. Current methodology uses reverse transcriptase polymerase chain reaction (RT-PCR) and genomic deoxyribonucleic acid (DNA) PCR techniques to detect NPM1 DIM. Although these methods are robust and relatively easy to perform they can be expensive, labour intensive and not universally available. Six major variants of NPM1 DIM (Types A-F) have been described all leading to frame shift. All six types share the same last five amino acids in the C-terminal.The aim of this study was to develop a robust flow cytometry methodology that could be used in the routine assessment of AML samples to determine the mutational state of NPM, using a commercially available polyclonal antibody against the mutated NPM1.
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