DNA damage-induced senescence reshapes transcriptomic and functional landscape of human neural progenitor cells

Ageing-related decline in hippocampal neurogenesis has been associated with cognitive impairment and neurodegenerative disease, yet experimentally tractable human models to study the underlying cellular and molecular mechanisms remain limited. Cellular senescence has emerged as a candidate driver of age-related tissue dysfunction, but its induction and consequences in human NPCs have not been well characterized. Here, we established a human in vitro model of NPC senescence using induced pluripotent stem cell-derived NPCs exposed to transient low-dose doxorubicin to activate the DNA damage response (DDR) while minimizing acute cytotoxicity. Doxorubicin-treated NPCs developed a stable senescent phenotype characterized by increased senescence-associated β-galactosidase activity, reduced proliferation, persistent DNA damage, and sustained induction of p21 and p16. Transcriptomic profiling revealed widespread senescence-associated remodeling, including activation of p53 and inflammatory programs and repression of cell cycle and DNA repair pathways. Senescent NPCs exhibited apoptosis resistance despite transcriptional priming of apoptotic pathways and underwent mitochondrial remodeling with a shift towards oxidative metabolism. In parallel, they acquired a senescence-associated secretory phenotype enriched in inflammatory, TGFβ-related and pro-angiogenic factors, and conditioned media from these cells promoted angiogenesis in vascular organoids. Importantly, key senescence-associated features were recapitulated in human hippocampal organoids, confirming the robustness of this paradigm in a three-dimensional neural context. Together, these findings establish a tractable human model of DDR-driven NPC senescence and identify senescence as a mechanism linking genotoxic stress to impaired progenitor function, metabolic rewiring, and paracrine niche remodeling relevant to hippocampal ageing and neurodegeneration.