Gene expression profiling and pathway analysis in acute myeloid leukaemia-normal karyotype patients

Acute myeloid leukaemia-normal karyotype (AML-NK) exhibits heterogeneity in expression profiles, influencing the treatment response and survival outcome. Transcriptome sequencing allows a comprehensive analysis of differentially expressed genes (DEGs) and dysregulated pathways in AML-NK, shedding light on the molecular mechanisms and their implications in patients’ management. DEG analyses utilising transcriptome sequencing were conducted using a customised DESeq2 pipeline on 51 AML-NK patients at diagnosis (DX), 12 AML-NK patients who attained first remission (CR1) and 12 healthy controls. The transcriptomic sequencing of AML-NK compared to healthy controls revealed 5,126 DEGs, comprising 85.8% coding genes and 14.2% non-coding elements across 37 pathway categories. The AML-NK DX versus CR1 identified 5,621 DEGs consisting of 84.7% coding genes and 15.3% non-coding elements affecting 20 categories of pathways. Gene set enrichment analysis in this study revealed consistent upregulation of proliferative pathways, including cell cycle and DNA replication. In contrast, immune-related pathways, such as cytokine-cytokine receptor interactions and MHC antigen presentation pathways, were downregulated. Overexpression of oncogenes ( FLT3, MYB, DNMT3B, and MYCN ) in DX vs CR1 samples reinforces their usefulness in minimal residual disease monitoring, especially in AML-NK with no genetic aberrations. These findings reiterate the known hallmarks of cancers and validate the transcriptomic dysregulation in the pathogenesis of AML-NK. The robustness of the transcriptome sequencing findings was confirmed by RT-qPCR validation of six genes that were not reported in AML-NK patients. The comprehensive analyses of pathways with dysregulation of a myriad of genes led to an understanding of AML-NK pathogenesis and highlighted the markers for minimal residual disease. In summary, this study performed the first transcriptome-wide analysis of AML-NK in a Malaysian cohort and underscored pathways that are candidates for therapeutic interventions.