Genetically Defined Acute Myeloid Leukemia Induced Pluripotent Stem Cell Clones Reveal Selective Impairment of...

Patient-derived induced pluripotent stem cells (iPSCs) provide a renewable model for studying acute myeloid leukemia (AML), but whether AML-iPSCs faithfully preserve the functional heterogeneity of genetically distinct AML clones remains unclear. Here, we functionally characterized a genetically defined AML-iPSC resource comprising 34 AML-iPSC lines from 11 AML patients, with complementary assays performed across partially overlapping subsets of lines. AML-iPSC lines were classified as aberration-positive (Ab⁺) or aberration-negative (Ab^-) based on retention of the clinically defining patient-specific leukemia-associated genetic abnormalities. Representative Ab⁺ and Ab^- AML-iPSC lines derived from donor hematopoietic cells exhibited comparable differentiation capacity across nonhematopoietic lineages, whereas Ab⁺ AML-iPSC lines demonstrated reduced hematopoietic differentiation, as determined by phenotypic analysis and functional colony-forming assays. Across overlapping subsets of AML-iPSC lines, these clone-specific hematopoietic phenotypes were conserved across four independent differentiation platforms, supporting that they reflect stable clone-specific properties rather than methodological artifacts. Residual hematopoietic progenitors derived from Ab⁺ AML-iPSC lines retained leukemic features, including persistence of leukemia-associated genetic abnormalities, immature blast-like morphology, and enhanced serial replating capacity indicative of self-renewal-associated properties. In a representative paired patient model, these progenitors also demonstrated persistent myeloid-skewed hematopoietic output following xenotransplantation. Together, these findings establish a renewable platform for mechanistic studies investigating how clonal heterogeneity contributes to disease evolution and therapeutic resistance in AML following pluripotent reprogramming.