Huaier Enhances the Antitumor Activities of Black Phosphorus and Alleviates Nephrotoxicity by Regulating Mitophagy and Autophagy

Black phosphorus (BP) has shown promise as a nanomaterial for cancer therapy, but its nephrotoxicity limits its use. To improve BP anti-tumor effects and reduced toxic side effects, this study presents a novel approach to cancer therapy by combining BP with the traditional Chinese medicine Trametes robiniophila Murr (Huaier) to improve the treatment outcome for pancreatic cancer, a highly fatal malignancy. In 2D/3D cell lines, patient-derived organoids (PDOs) and mouse model, Huaier extraction is demonstrated to the antitumor effects of BP and mitigates BP-induced nephrotoxicity. RNA-sequencing analysis revealed that the combination of BP and Huaier enriched the mitophagy and ferroptosis pathways. While BP activates the mitophagy pathway to recycle damaged mitochondria, Huaier accelerates BP-induced mitochondrial dysfunction, leading to a mitochondrial burst and ultimately activating ferroptosis in PAAD cells. Furthermore, Huaier mitigates BP-induced nephrotoxicity and recovers cell viability by modulating the level of autophagy in proximal tubule epithelial cells. By synergizing BP with Huaier, we have achieved more potent antitumor activities with lower nephrotoxicity. These findings provide a promising strategy for treating PAAD in in vitro and in vivo models. Overall design: Comparative gene expression profiling analysis of RNA-seq data for MIAPaCa-2 cells treated with BP or BP+Huaier