Differentiation Therapy in Acute Myeloid Leukemia: Advances in Phenotypic Screening and CRISPR-based Functional Genomics

Acute myeloid leukemia (AML) is a clinically and genetically heterogeneous malignancy marked by a differentiation block in myeloid progenitors. Despite advances in molecular-targeted therapies, long-term outcomes remain poor, underscoring the need for novel treatment strategies. Differentiation therapy, exemplified by the success of all-trans retinoic acid in acute promyelocytic leukemia, offers a compelling alternative to cytotoxic approaches. This review highlights recent advances in phenotypic screening strategies-including high-throughput compound libraries, computational tools, and CRISPR-based functional genomics-which have revealed novel differentiation inducers, genes, and regulatory pathways. Notably, recent phenotypic screening studies identified Triciribine (TCN), an AKT inhibitor, as a differentiation inducer in AML cells. In addition, CRISPR loss- and gain-of-function screens have uncovered key regulators of differentiation, such as transcriptional (KAT6A), metabolic (NMNAT1, GLUT1), and post-transcriptional (ZFP36L2, YTHDC1) effectors. Emerging computational methods, such as the Lineage Maturation Index and single-cell data integration, further enhance target prioritization and translational relevance. However, differentiation therapy outside APL has shown variable and often incomplete clinical success, frequently limited by partial maturation and context-dependent responses. Together, these approaches reveal novel therapeutic vulnerabilities in AML and support the development of differentiation-based strategies for a broader range of patients.