DDO1002, an Nrf2–Keap1 inhibitor, improves impairedhaematopoiesis by reducing ROS production and promoting Nrf2-mediated antioxidative response

Oxidative stress reduces haematopoietic stem cell (HSC) function with age, and increased reactive oxygen species (ROS) levels promote DNA damage, cell senescence, and haematopoietic dysfunction. DDO1002, an effective Keap1–Nrf2 inhibitor, may regulate antioxidant gene expression, however, its ability to alleviate impaired haematopoiesis following total body irradiation (TBI) or ageing remains unclear. Through both cellular and mouse models, we showed that DDO1002 upregulated Nrf2, activated the Nrf2-mediated antioxidant response element (ARE) signalling pathway, reduced intracellular ROS levels and delayed cellular senescence. Moreover, DDO1002 reduced DNA damage and HSC apoptosis, thereby increasing the number of HSCs, lymphoid-primed MPP4 cells, and B cells in peripheral blood. Therefore, DDO1002 alleviates TBI-induced haematopoietic injury. Similarly, DDO1002 treatment activated the expression of Nrf2 and Nrf2-mediated ARE signalling pathways in the bone marrow cells of naturally ageing mice. B and T cell proportions in the spleen were increased, as was the colony-forming ability of bone marrow cells. Single-cell sequencing analysis revealed that DDO1002 treatment attenuated intracellular inflammatory signalling pathways and reduced the ROS pathway in aged HSCs, suggesting an ability to restore aged-HSC viability. Thus, DDO1002 effectively activated Nrf2 to delay cell senescence and improved impaired haematopoiesis via the Nrf2–ARE pathway, showing potential for treating age-related haematopoietic disorders. Analysis gene expression and pathways affected by Nrf1-Keap1 inhibitor DDO1002 in each cell populations of bone marrow in single-cell level using scRNA-seq.