Acute Myeloid Leukemia Cells Research Articles

Mixed-lineage leukemia (MLL) rearrangements and Nucleophosmin-1 (NPM1) mutations are associated with acute leukemias whose pathogenesis is critically influenced by protein-protein interactions between menin and MLL. We hypothesized that targeting the menin-MLL interaction using DS-1594b and blocking the antiapoptotic BCL-2 protein using venetoclax may promote differentiation and enhance eradication of MLL-rearranged and NPM1-mutated leukemias models. We treated acute myeloid leukemia (AML) cell lines with MLL rearrangements, NPM1 mutations, other leukemias and primary samples from AML patients with venetoclax alone, DS- 1594b alone, and their combination. We measured proliferation, viability, apoptosis, and differentiation using a variety of cellular assays, Western blotting, and BH3 profiling. Treatment with DS-1594b and venetoclax exerted significant synergy, resulting in enhanced differentiation and inhibited proliferation across several cell lines. In the NPM1-mutated AML PDX model, DS- 1594b single-agent treatment significantly extended survival. Importantly, compared with DS- 1594b monotherapy, the combination of DS-1594b and venetoclax more profoundly reduced leukemic burden and prolonged mouse survival. Menin inhibition was the primary driver of transcription changes in this model and impacted the expression of antiapoptotic regulators, providing a mechanistic explanation for the synergy observed between these drugs. Overall, we observed synergistic effects on differentiation induction and proliferation inhibition, both in vitro and in vivo. Together, our studies underscore the promise of this combination strategy as a novel therapeutic approach for improving treatment outcomes in patients with these specific genomic alterations.

The integration of chimeric antigen receptor (CAR) therapies with precision medicine holds potential to impact the treatment landscape for acute myeloid leukemia (AML). Genetic mutations play a role in the efficacy of CAR-T and CAR-NK cells, influencing their crucial role in determining the effectiveness of these cells, as well as their proliferation, persistence, resistance, and safety. This review examines how mutations in FLT3, DNMT3A, NPM1, TP53, TET2, gene fusions involving RUNX1 and KMT2A and other key genes modulate CAR-based immunotherapies, highlighting both vulnerabilities and resistance mechanisms. Recent findings demonstrate that mutations in genes such as DNMT3A and NPM1 enhance antigen expression, thereby improving CAR targeting. In contrast, mutations in TP53 drive immune escape and resistance to therapy. Understanding these mutation-specific effects is essential for tailoring CAR therapies to individual patients, optimizing efficacy while minimizing toxicity. By leveraging genomic profiling and personalized engineering approaches, CAR therapies can be refined to overcome resistance and enhance precision in AML treatment. Future research should focus on integrating multiomic data to develop mutation-adapted CAR strategies, ensuring that patients receive the most effective and personalized immunotherapy.

Guanine nucleotides drive ribosome biogenesis and glycolytic reprogramming in acute myeloid leukemia stem cells.

Background Acute myeloid leukemia (AML)-M2b with t(8;21)(q22;q22)/RUNX1::RUNX1T1 (AML1-ETO) is associated with a high risk of relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Post-transplant lymphoproliferative disorder (PTLD), particularly involving the central nervous system (CNS), confers a poor prognosis. Although CD19 chimeric antigen receptor T-cell (CAR-T) therapy is established in B-cell malignancies, its application in acute myeloid leukemia (AML) or CNS-PTLD has rarely been reported. Case A 24-year-old male with AML-M2b showed persistent RUNX1::RUNX1T1 (AML1-ETO) positivity after allo-HSCT. He developed an extramedullary relapse (presacral mass) at 7 months, followed by CNS-PTLD with limb palsy at 9 months post-HSCT. The disease subsequently progressed to bone marrow relapse (RUNX1::RUNX1T1 94.42%, MRD >5%). Intervention Given the co-expression of CD19 on both the AML and PTLD cells, the patient was treated with donor-derived CD19 CAR-T cells. He experienced manageable grade 1 cytokine release syndrome (CRS) and grade 3 Immune Effector Cell-Associated Neurotoxicity Syndrome (ICANS). Outcomes The patient achieved a complete response (CR) with negative MRD, disappearance of the fusion gene, reduction of PTLD and extramedullary lesions, and recovery of limb strength. Conclusion This case demonstrates the efficacy and feasibility of CD19 CAR-T therapy for concomitant post-transplant AML-M2b relapse and CNS-PTLD, leveraging their shared CD19 expression. It provides clinical evidence that targeting a shared antigen with a single CAR-T product can effectively treat heterogeneous malignancies, offering a promising new strategy for such complex cases.

DNA nanostructure-enabled functional constructs have shown potential to improve healthcare outcomes by offering advanced disease diagnostic and therapeutic strategies. Translating this potential of DNA nanostructure-based constructs to real life applications relies on maintaining and enhancing the structural integrity and functions of the surface-anchored moieties. In this study, we explored the possibility of utilizing choline dihydrogen phosphate (CDHP) solution, an aqueous solution of ionic liquid, to assemble DNA nanostructures of different sizes and complexities with enhanced biostability and ligand binding affinity. We show successful formation of the DNA nanostructures in aqueous CDHP solution using gel electrophoresis, atomic force microscopy (AFM), and circular dichroism (CD). Biostability assays reveal that the aqueous CDHP solution may provide passive protection to DNA nanostructures against DNase I and human serum for up to 48 h. We also demonstrate that this enhanced biostability arises both from the structural conformation imparted during CDHP-mediated folding and from the presence of free CDHP ions in the solution. Notably, removal of free ions reduced the passive protection effect, but did not eliminate it, indicating the contribution of both folding and surrounding free ions. Using flow cytometry and surface plasmon resonance assays, we show that the presence of aqueous CDHP solution can enhance the binding of aptamer-functionalized DNA nanostructures to specific receptors on acute myeloid leukemia (AML) cells. Our strategy of using ionic liquid solution for one-pot preparation with enhanced stability and functionality offers a robust, simpler and faster alternative for DNA nanostructure-based constructs.

Acute myeloid leukemia (AML) is a clinically aggressive hematologic malignancy characterized by high relapse rates and treatment resistance, highlighting the need for novel biomarkers to improve clinical outcomes. In this study, we explore the roles of nuclear receptor-interacting protein 1 (NRIP1) in AML, focusing on its associations with tumor progression and immune infiltration. Analysis of public AML gene expression datasets reveals that NRIP1 expression is significantly increased in AML patients. Those with high NRIP1 expression have markedly shorter overall survival than those with low expression. Furthermore, NRIP1 expression is significantly associated with the infiltration of diverse immune cells, including B cells, dendritic cells, T cells, mast cells, eosinophils, and T helper cells, suggesting that NRIP1 may be a regulator of immune cell infiltration. Functional enrichment analysis indicates that NRIP1 and its interacting partners are involved in tumorigenesis, immune microenvironment remodeling, and metabolic reprogramming. Survival analysis confirms the prognostic value of NRIP1. Importantly, functional validation in AML cell lines confirms that NRIP1 knockdown suppresses proliferation and induces apoptosis. Our study identifies NRIP1 as a multifaceted regulator that promotes AML by driving tumor progression, regulating immune cell infiltration, and modulating ferroptosis, highlighting its role as a novel prognostic biomarker.

B-cell lymphoma-2 (BCL2) is an anti-apoptotic molecule. Previous studies on its function focused on leukemic cells in acute myeloid leukemia (AML), whereas its effect in lymphocytes is unclear. We performed multi-parameter flow cytometry to test BCL2, immune checkpoint and cytotoxic molecules on T and NK cells in bone marrow samples of 80 newly diagnosed adult AML patients and 7 healthy donors (HDs). BCL2 expression was evaluated on T-cell differentiation subsets in a subset of 28 AML patients and 6 additional HDs. AML patients had lower BCL2+ frequencies on CD4+T and CD8+T cells and similar frequency on NK cells than HDs (p = 0.001, 0.001 and 0.34). Effector memory T cells had lower BCL2+ frequency than naïve T, stem central memory T and central memory T cells (all p < 0.05) in AML. Lower BCL2+ frequency on NK cells predicted higher 1-course complete remission rates (p = 0.016). Lower BCL2+ frequency on T cells was related to poor relapse-free survival (all p < 0.001) and that on CD4+T cells was an independent predictor (p = 0.008). BCL2+ cells had lower PD1+ and TIM-3+ frequencies than BCL2− cells on CD4+T cells, and lower PD1+ but higher TIM-3+, perforin+ and Granzyme B+ frequencies compared to BCL2− cells on CD8+T and NK cells (all p < 0.05). BCL2 expression on T and NK cells in diagnostic bone marrows are associated with treatment response and outcome in adult AML, which might be relevant to their association with immune checkpoint and cytotoxic molecules expression.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-22331-5.

DNA damage agents trigger iNKT cell-mediated elimination of AML cells through activation of NF-κB/HLA-DRB6/CD1d pathway.

Acute myeloid leukemia (AML) is a heterogeneous disease with an unmet need for novel therapeutic drugs. Previous studies have reported the upregulation of diacylglycerol kinases (DGKs) in AML. This study investigated the effects of ritanserin, a DGKα-specific inhibitor, and DGKζ-IN4 or BAY 2965501, DGKζ-selective inhibitors, on a panel of AML cell lines. Ritanserin induced apoptotic cell death across all tested models, whereas DGKζ inhibitors triggered both apoptosis and necrosis to variable extents, with HL-60 cells being the most responsive to both compounds. Drug sensitivity did not correlate with DGKα or DGKζ expression levels, indicating that additional factors may influence cellular susceptibility. THP-1 proteomic profiling revealed that ritanserin broadly downregulated proteins involved in antigen presentation, cell cycle and metabolism, while BAY 2965501 affected a smaller and distinct but functionally similar protein subset, implying different mechanisms of action. Gene silencing confirmed AML cell line-specific dependence on DGK isoforms: HEL cells were sensitive to DGKα knockdown, HL-60 to DGKζ silencing, whereas K562 and THP-1 were resistant to both. These findings indicate that DGKs targeting can effectively reduce AML cell viability. However, AML heterogeneity and the limited selectivity of current inhibitors underscore the need for predictive biomarkers and combinatorial strategies to translate DGK inhibition into effective therapy.

Targeting serine metabolism vulnerability in omipalisib-resistant acute myeloid leukemia with phosphoglycerate dehydrogenase inhibitors.

KIF11 promotes AML progression, and its inhibition by SB-743921 suppresses disease advancement through mitotic G2/M phase arrest.

Precision targeting of CXCR4+ leukemia cells by a humanized MMAE-nanoconjugate in an AML mouse model.

Targeting acute myeloid leukemia resistance with two novel combinations demonstrate superior efficacy in TP53, HLA-B, MUC4 and FLT3 mutations.

We examined the role of SLFN12, a member of the Schlafen (SLFN) family of interferon-regulated genes and proteins in leukemogenesis and its potential as a therapeutic target in acute myeloid leukemia (AML). We explored the effects of velcrins, a class of small molecules able to modulate SLFN12 biological activity, on AML cells. Velcrin treatment of AML cells stabilized SLFN12 and promoted SLFN12 complex formation with phosphodiesterase 3A (PDE3A) or phosphodiesterase 3B (PDE3B). Such effects were associated with growth inhibitory and pro-apoptotic responses, as well as potent suppressive effects on leukemic cell growth. In addition, velcrin treatment suppressed clonogenic capacity of primitive leukemic progenitors and significantly extended survival in a mouse AML xenograft model. Taken together, these findings establish an important role of SLFN12 in leukemogenesis and raise the potential for the use of velcrins as a therapeutic strategy for AML.

Despite therapeutic advances, acute myeloid leukemia (AML) remains associated with high relapse rates and poor outcomes, especially in elderly or unfit patients. Allium sativum is widely known for its medicinal properties, but its anti-leukemic effects have not been fully explored. This study investigated the cytotoxic and pro-apoptotic potential of an ethyl acetate extract of Allium sativum (EAEAS) using ex vivo mononuclear cells from thirteen AML patients. Cytotoxicity was assessed via MTT assay, while mechanisms of cell death were analyzed using annexin V/propidium iodide staining, DNA fragmentation, mitochondrial depolarization, and caspase 3/7 activation. EAEAS induced a dose-dependent decrease in cell viability, with variable IC₅₀ values across samples. Apoptosis was confirmed through mitochondrial dysfunction, sub-G1 accumulation, and caspase activation. Additionally, we evaluated the effect of EAEAS on two LSC subpopulations using multicolor flow cytometry. Although no statistically significant changes were observed, some patients showed a decrease in LSC frequencies after treatment, suggesting possible selective activity. These findings indicate that EAEAS triggers intrinsic apoptosis in AML blasts and may exert partial activity on stem-like compartments, warranting further evaluation as an adjuvant therapy in AML.

Dipeptidyl peptidase 9 (DPP9) is a key regulator of pyroptosis in leukocytes. DPP9-targeting inhibitors have been reported to selectively induce pyroptosis in human acute myeloid leukemia (AML) cells and work synergistically with non-nucleoside reverse transcriptase inhibitors (NNRTIs) to kill HIV-1-infected lymphocytes. Here, we report structure-activity relationship data for a novel series of low nanomolar DPP9 inhibitors with unprecedented pyroptosis-inducing potency and kinetics. They have substantial DPP9-to-DPP8 selectivity and full selectivity over other related peptidases, including DPP4. The selected compound 6e was administered to healthy rats and demonstrated high oral bioavailability, along with a long in vivo and microsomal half-life. Finally, we also investigated the pyroptosis induction potential in HIV-1-infected T-lymphocytes. These new compounds have the potential to become important research tools and support further progress in DPP9's therapeutic potential.

Herein, we describe experiments showing that liposomal spherical nucleic acid (SNA) constructs comprised of 5-fluorouracil (5-Fu) are selectively taken up by myeloid cells, including acute myeloid leukemia (AML) cells, and act as chemotherapeutics. Specifically, SNAs with biocompatible phospholipid-based liposome cores and oligonucleotides consisting of 10 units of the chemically interconnected 5-fluoro-2'-deoxyuridine, were designed and synthesized. These oligonucleotides are modified in the 3' position with hexaethylene glycol and cholesterol end groups, which allow them to be anchored to the liposomal cores. Small-molecule drugs like 5-Fu are conventionally delivered via encapsulation in the liposome interior, but restructuring them in the form of an SNA increases their cellular uptake by up to 12.5-fold and enables preferential delivery to AML cell lines. Up to 4 orders of magnitude enhancement in cell killing was observed using chemotherapeutic SNAs compared to the free small molecule 5-Fu in vitro. In a human AML model based on immunodeficient mice, the chemotherapeutic SNA exhibited 59-fold better antitumor efficacy than 5-Fu and improved AML-associated hematological markers without observable side effects. This study highlights the advantages in potency that can be realized when chemotherapeutics are integrated within SNAs, and it underscores how the structure of nanomedicine can profoundly impact both chemotherapeutic delivery and cell targeting.

The XPO7-NPAT axis represents key vulnerabilities in TP53-mutated acute myeloid leukemia.

Menin inhibition enhances graft-versus-leukemia effects by T-cell activation and endogenous retrovirus induction in AML.

Multiselective RAS(ON) inhibition targets oncogenic RAS and overcomes RAS-mediated resistance to FLT3i and BCL2i in AML.