Xiuyun Wu, Jingxin Zhang
Objective Acute myeloid leukemia (AML) arises from blocked myeloid differentiation and subsequent accumulation of immature myeloid progenitors. Leukemia stem cells (LSCs) sustain AML hierarchical architecture via persistent self-renewal capacity and resistance to terminal differentiation. Elucidating the molecular disruption of differentiation programs is critical for illuminating leukemogenesis mechanisms and developing targeted therapeutic strategies. This review aims to systematically summarize the multi-layered molecular mechanisms governing LSC differentiation arrest in AML, and highlight the application value of multi-omics integrative analysis in mechanistic exploration and therapeutic translation. Methods This review comprehensively collected and integrated recent research advances focusing on AML LSC differentiation regulation. We systematically summarized updated findings regarding LSC hierarchical characteristics, phenotypic plasticity, transcriptional regulatory networks, epigenetic barriers, aberrant signaling pathways, and metabolic reprogramming. Furthermore, we concluded high-resolution research progress achieved by integrated multi-omics technologies including genomics, transcriptomics, epigenomics, proteomics, metabolomics and single-cell multi-omics approaches in AML research. Results Multiple molecular layers collaboratively drive differentiation blockade of AML LSCs. Core myeloid transcription factors and oncogenic fusion proteins remodel lineage-specific gene expression and balance LSC self-renewal and differentiation. Abnormal epigenetic modifications, including DNA/RNA methylation, histone alteration, chromatin remodeling and 3D genome organization, establish stable epigenetic barriers to block LSC terminal differentiation. Dysregulated signaling cascades (FLT3, Wnt/β-catenin, PI3K/AKT/mTOR) and disordered metabolic programs (mitochondrial function, redox homeostasis, proteostasis) serve as key regulators of LSC fate decisions. Multi-omics integrative strategies precisely dissect AML cellular heterogeneity, clonal evolution and abnormal differentiation trajectories at high resolution. Conclusion Differentiation blockade is the core biological feature of AML LSCs. Current multi-omics studies have profoundly uncovered the transcriptional, epigenetic, signaling and metabolic mechanisms underlying LSC differentiation arrest. Targeting LSC differentiation via epigenetic modification, metabolic intervention and omics-guided precision strategies provides promising therapeutic directions for AML. Nevertheless, multiple challenges remain in clinical transformation of differentiation-based therapies. Further in-depth multi-omics research is required to advance the precise targeting and clinical application of AML LSC differentiation.