Differentiation Therapy in Acute Myeloid Leukemia: Advances in Phenotypic Screening and CRISPR-Based Functional...

Acute myeloid leukemia (AML) is a clinically and genetically heterogeneous malignancy characterized by a differentiation block in myeloid progenitors. Although advances in molecular targeted therapies have improved outcomes in selected subgroups, long-term prognosis remains poor for many patients. Differentiation therapy, exemplified by the success of all-trans retinoic acid in acute promyelocytic leukemia (APL), represents an alternative therapeutic paradigm that aims to overcome the differentiation blockade rather than directly inducing cytotoxicity.

This review summarizes recent advances in phenotypic screening and CRISPR-based functional genomics that have contributed to the discovery of novel differentiation-inducing strategies in AML. High-throughput phenotypic screening approaches using compound libraries, computational tools, and integrative transcriptomic analyses have identified several candidate differentiation inducers. For example, triciribine, an AKT inhibitor, has been identified as a differentiation-inducing compound in AML models. In parallel, CRISPR loss- and gain-of-function screens have uncovered multilayered regulatory networks governing AML differentiation, including transcriptional regulators (e.g., KAT6A), metabolic dependencies (e.g., NMNAT1, glucose transporter type 1), and post-transcriptional regulators (e.g., ZFP36L2, YTHDC1). Emerging computational approaches, such as the Lineage Maturation Index and single-cell data integration, further enhance target prioritization and improve the translational relevance of screening results. Despite these advances, differentiation therapy outside APL remains challenging due to partial maturation, context-dependent responses, and AML heterogeneity.

Recent advances in phenotypic screening and CRISPR-based functional genomics have expanded our understanding of the molecular mechanisms governing AML differentiation and have revealed novel therapeutic vulnerabilities. Integration of these discovery platforms with computational and single-cell approaches may facilitate the development of differentiation-based strategies for a broader spectrum of AML patients.