A novel human acute myeloid leukemia cell line SDEY-AML1 with KMT2A: MLLT3, IKZF1: EVX1 fusions exhibits high tumorigenicity in NSG mice.

Human leukemia cell lines play a crucial role in leukemia research, particularly in understanding the pathogenesis of acute leukemia and developing novel therapeutic strategies. We here describe the generation of a novel monocytic cell line SDEY-AML1, harboring alterations in KMT2A::MLLT3, IKZF1::EVX1, ETV6, and TP53 double mutations, and its biological characteristics. We established a cell line, SDEY-AML1, from the mononuclear cells isolated from the bone marrow of a patient with acute myeloid leukemia (AML-M5). Comprehensive morphological and cytogenetic characterization of SDEY-AML1 cells was performed using cytochemical staining, flow cytometry, and karyotype analysis. The molecular features of the cell line were identified by transcriptomic analysis and reverse transcription-polymerase chain reaction (PT-PCR). The proliferative capacity of the SDEY-AML1 cells was assessed using a clonogenic assay. And we conducted drug sensitivity experiments on SDEY-AML1 to tested its sensitivity to commonly used chemotherapy drugs and targeted drugs. Additionally, we created an in vivo tumor model of SDEY-AML1 cells by injecting 1×107 cells subcutaneously in NSG mice via the tail vein. The SDEY-AML1 cells were maintained in continuous culture for one year without the addition of exogenous cytokines. Cytochemical assays revealed that the cells were positive for α-naphthyl acetate esterase reactivity that was partially inhibited by sodium fluoride but negative for peroxidase and periodic acid-Schiff staining. Flow cytometry showed the expression of myeloid markers in SDEY-AML1 cells. Karyotypic analysis revealed a t(9;11) (p22;q23) translocation in SDEY-AML1 cells, mirroring the chromosomal aberrations identified in the primary leukemia cells of the patient. Genetic profiling confirmed the presence of alterations in KMT2A::MLLT3, IKZF1::EVX1, ETV6, and TP53 genes. SDEY-AML1 cultures were negative for EBV and mycoplasma. NSG mice showed tumor infiltration within 40-50 days of tail vein injection of SDEY-AML1 cells. The new AML cell line, SDEY-AML1, harbored KMT2A::MLLT3, IKZF1::EVX1, ETV6 gene alterations, and TP53 double mutations. These cells were highly tumorigenic in NSG mice. This cell line has applications in the research and development of new agents targeting KMT2A::MLLT3, IKZF1::EVX1-associated leukemia, making it a valuable tool in the study of leukemogenesis.