Normal karyotype acute myeloid leukemia (NK‐AML) is a highly heterogeneous malignancy that resides within a complex immune microenvironment, complicating efforts to reveal the interaction between leukemia cells and immune cells. Understanding tumor‐infiltrating T cells is crucial to the advancement of immune therapies and the improvement of the prognosis for NK‐AML patients. We performed single‐cell RNA sequencing on bone marrow cells from 5 NK‐AML (M4/M5) patients and 1 normal donor and paired single‐cell T cell receptor (TCR) sequencing on single T cells. As a result, we identified 8 T cell clusters based on the gene expression characteristics of each subset in NK‐AML and described their developmental trajectories. In NK‐AML patients, specific clusters, such as mucosal‐associated invariant T cells (MAITs), were preferentially enriched and potentially clonally expanded. These transcriptome and TCR data analyses provide valuable insights and rich resources for understanding the immune environment of NK‐AML.

Normal karyotype acute myeloid leukemia (NK-AML) is a heterogeneous hematological malignancy that contains a minor population of self-renewing leukemia stem cells (LSCs), which complicate efforts to achieve long-term survival. We performed single-cell RNA sequencing to profile 39,288 cells from 6 bone marrow (BM) aspirates including 5 NK-AML (M4/M5) patients and 1 healthy donor. The single-cell transcriptome atlas and gene expression characteristics of each cell population in NK-AML (M4/M5) and healthy BM were obtained. In addition, we identified a distinct LSC-like cluster with possible biomarkers in NK-AML (M4/M5) and verified 6 genes using qRT‒PCR and bioinformatic analyses. In conclusion, we utilized single-cell technologies to provide an atlas of NK-AML (M4/M5) cell heterogeneity, composition, and biomarkers with implications for precision medicine and targeted therapies.

Normal karyotype acute myeloid leukemia (NK-AML) is a heterogeneous hematological malignancy that composes a minor of self-renewing leukemia stem cells (LSC), complicating efforts to achieve long-term survival. We performed single-cell RNA sequencing to profile 39288 cells from 6 bone marrow aspirates, including 5 NK-AML(M4/M5) patients and 1 healthy donor. The single cell transcriptome atlas and the gene expression characteristics of each cell population in NK-AML (M4/M5) and healthy bone marrow (BM) were obtained. Besides, we identify a distinct LSC-like cluster with possible biomarkers in NK-AML(M4/M5)and verified 6 genes using qRT-PCR analysis and bioinformatics analysis. In conclusion, we provide single-cell technologies and an atlas of NK-AML(M4/M5) cell heterogeneity, composition, and biomarkers with implications for precision medicine and target therapies.