ROS-NRF2 axis coordinates concentration-graded differentiation-to-PANoptosis switching in a phthalide combination...

In the context of multimodal therapeutic strategies for acute myeloid leukemia (AML), the coordinated regulation of cell death and differentiation pathways has emerged as a groundbreaking research direction.

the study aims to investigates the synergistic anti-AML mechanisms of two phthalide derivatives from Chuanxiong Rhizoma, Z-Ligustilide (Z-LIG, a potent pro-apoptotic agent) and Senkyunolide A (Sen-A, a differentiation inducer).

The anticancer activity of Z-LIG and Sen-A, and their synergistic effects, were evaluated using the SRB assay. The cells underwent differentiation and PANoptosis (the forms of cell death with apoptosis, pyroptosis and necroptosis at same time), as evidenced by alterations in cell morphology observed through flow cytometry and transmission electron microscopy (TEM). The upregulation of NRF2 in both differentiation and PANoptosis was confirmed through the use of inhibitors (ML385, Dicoumarol), and the examination of ROS levels in differentiation and PANoptosis through flow cytometry. Finally, the findings were validated in an AML-NOD/SCID mouse model.

In vitro experiments revealed that Z-LIG/Sen-A combination triggers concentration -dependent dual effects in AML cells: low- concentration treatment promotes differentiation via H₂O₂-mediated reprogramming of iron homeostasis, whereas high- concentration administration induces PANoptosis through ASC-RIPK3-caspase-8 inflammasome assembly driven by H₂O₂ accumulation. Mechanistically, a bidirectional regulatory network between ROS and NRF2 orchestrates both iron metabolism and PANoptosis pathways. In AML-NOD/SCID murine models, the combination therapy significantly attenuated leukemia progression and activated the bone marrow NRF2/ROS axis without inducing nephrotoxicity.

The study pioneers the elucidation of phthalide-induced "differentiation-to-PANoptosis" fate switching governed by H₂O₂ dosage gradients and NRF2/ROS crosstalk, providing a mechanistic framework for developing precision TCM-based combinatorial therapies against hematological malignancies.