Single-cell transcriptomic characterization of a human MDS hematopoietic dataset and in vitro validation of apoptosis-related venetoclax resistance in human MDS-L cells.

BACKGROUND

Myelodysplastic syndromes (MDS) are heterogeneous clonal hematopoietic disorders in which dysregulated apoptosis, oxidative stress, and adaptive survival signaling contribute to disease persistence and therapeutic resistance. Venetoclax (VEN) targets BCL-2-dependent survival, but resistance is frequently accompanied by compensatory survival programs. Whether ferroptosis-associated transcriptional states are dynamically remodeled during VEN treatment and participate in this adaptive phenotype remains incompletely understood.

METHODS

We analyzed two independent human MDS single-cell RNA-sequencing cohorts together with an in vitro VEN-resistant MDS-L model. GSE241417 was used to characterize longitudinal and cell-state-specific ferroptosis-associated transcriptional patterns, whereas GSE180298 provided independent single-cell validation. Longitudinal module scores were summarized across major hematopoietic states and treatment stages. Apoptosis/proliferation- and ferroptosis-related genes were further examined by qRT-PCR in parental and VEN-resistant MDS-L cells.

RESULTS

Single-cell analysis revealed marked heterogeneity of ferroptosis-associated transcriptional programs across MDS hematopoietic populations. Longitudinal trajectories in GSE241417 demonstrated nonuniform, treatment-stage- and cell-state-dependent remodeling of ferroptosis balance, inflammatory context, and survival-associated programs. The independent GSE180298 cohort reproduced major disease-associated ferroptosis-related transcriptional differences. In VEN-resistant MDS-L cells, MCL1, BCL2L1, MKI67, GPX4, SLC7A11, and NFE2L2 were increased, whereas BCL2, DDIT3, ACSL4, and NCOA4 were decreased, indicating coordinated adaptation of apoptotic and ferroptosis-related pathways.

CONCLUSION

This study reveals cell-state-dependent and temporally dynamic ferroptosis-associated remodeling in MDS and identifies a coordinated apoptosis-redox transcriptional program associated with VEN resistance. The integration of longitudinal single-cell analysis, independent single-cell validation, and targeted qRT-PCR supports ferroptosis-defense pathways as a biologically relevant component of VEN-resistant adaptation and provides a rationale for future combination strategies that address both apoptotic dependence and redox-state plasticity.