Decoding the hierarchical transcription factor network in hematopoiesis: from stem cell fate control to translational therapeutic opportunities

Hematopoietic stem cell (HSC) self-renewal and multilineage differentiation form the functional core of lifelong hematopoietic homeostasis. A critical translational bottleneck in clinical hematology is the lack of strategies for robust, functional ex vivo expansion of human HSCs, which severely limits their use in clinical hematopoietic transplantation and regenerative medicine. While transcription factor (TF) networks govern HSC fate decisions, current knowledge focuses largely on individual TFs and isolated signaling axes. A hierarchical framework that integrates their cooperative logic, dose-dependent regulatory effects, and pathological cascades in human hematopoiesis remains lacking. This review integrates contemporary human and murine evidence to construct a four-tiered regulatory architecture that governs HSC ontogeny, homeostasis, and lineage commitment. We describe cooperative and antagonistic interactions among core TF families (ETS, HOX, GATA, C/EBP, RUNX1, STAT) within a stratified hierarchy: (i) an apex maintenance tier preserving stem cell identity, (ii) a guardian tier sustaining HSC quiescence and lineage priming potential, (iii) a lineage priming and stress response tier integrating microenvironmental signals, and (iv) discrete lineage-specific execution modules driving terminal differentiation. We integrate emerging single-cell multi-omics evidence to distinguish findings directly demonstrated in human cells from those extrapolated from model organisms, highlight interspecies differences between murine and human HSC regulation, and propose that systemic dysregulation of this hierarchical network may explain human hematopoietic failure and leukemogenesis more fully than isolated single-factor mutations. Network-guided intervention targeting tier-specific regulatory nodes is presented as a promising but largely conceptual direction that requires systematic preclinical and clinical validation. Integrating single-cell multi-omics profiling with this human-refined hierarchical model provides a framework for identifying novel targets for ex vivo human HSC expansion and developing network-guided, precision therapeutic strategies for hematological malignancies and regenerative medicine.