Characterisation of mutations in cytogenetically normal acute myeloid leukaemia patients with triple-negative FLT3-ITD/ NPM1/CEBPA subgroup using targeted next-generation sequencing

Zahidah Abu Seman¹, Nor Rizan Kamaluddin¹, Noor Atiqah Fakharuzi¹, Ermi Neiza Mohd Sahid¹, Ezalia Esa¹, and Yuslina Mat Yusoff^1*

1. Haematology Unit, Cancer Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, 40170 Shah Alam, Selangor, Malaysia

*Corresponding author: Yuslina Mat Yusoff. This email address is being protected from spambots. You need JavaScript enabled to view it.. Tel: +603-33627869

CITATION: Abu Seman Z, Kamaluddin NR, Fakharuzi NA, Mohd Sahid EN, Esa E, Mat Yusoff Y. Characterisation of mutations in cytogenetically normal acute myeloid leukaemia patients with triple-negative FLT3-ITD/ NPM1/CEBPA subgroup using targeted next-generation sequencing. International Medical Research Journal. 2025 May 30;11(1):31–41. https://doi.org/10.63719/imrj.2025.11.01.003

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ABSTRACT
Genetic aberrations play a pivotal role in acute myeloid leukaemia (AML) pathogenesis and clinical outcomes, with mutations in key genes such as NPM1, FLT3, and CEBPA well-established markers for the disease prognosis. Despite advancements in molecular diagnostics, a comprehensive analysis of the mutational landscape in AML is still needed using advanced techniques. This study aimed to analyse the mutational landscape of cytogenetically normal AML (CN-AML) with the triple-negative FLT3-ITD/NPM1/CEBPA subgroup using targeted NGS techniques. Targeted next-generation sequencing (NGS) was employed to comprehensively explore the mutational spectrum of CN-AML in a cohort of 14 patients. In this study, six putative pathogenic or likely pathogenic variants, as well as seven variants of uncertain significance (VUS), were detected in nine CN-AML patients. Among these patients, a single mutation was observed in six cases, two mutations in one case, and three mutations in two cases each. DNMT3A and RUNX1 were present in three cases each, followed by SRSF2 and NRAS, which were found in two cases each. Additionally, CEBPA, TET2, TP53, and U2AF1 were each observed in one case. Targeted NGS analysis revealed a heterogeneous mutational spectrum in CN-AML patients with recurrent alterations, highlighting their potential impact on disease progression. These findings improve our understanding of the genetic landscape of CN-AML and may contribute to refining prognostic classification and risk stratification.

KEYWORDS: acute myeloid leukaemia, next-generation sequencing, NPM1 gene, FLT3 gene, CEBPA gene