Hippo-YAP signaling alleviates copper-induced mitochondrial dysfunction and oxidative damage via the ATOX1-PPA2 pathway.

Hippo signaling plays a crucial role in the cellular response to various stressors, such as mechanical stress, metabolic stress, and hypoxic stress. However, its physiological significance in copper (Cu) stress remains poorly understood. Here, we demonstrated aberrant activation of Hippo-YAP signaling in sheep pancreas and pancreatic organoids exposed to excessive Cu, accompanied by significant pathological changes, elevated levels of oxidative stress, and impaired mitochondrial structure and function. The inhibition of Hippo signaling or overexpression of YAP protected against Cu-induced damage by improving mitochondrial function and maintaining cellular Cu homeostasis. YAP interacted with TEAD and upregulated the expression of Cu chaperone ATOX1, a key regulator of intracellular Cu homeostasis. ATOX1 restored mitochondrial function under Cu stress by reducing mitochondrial superoxide levels, increasing ATP production and mitochondrial membrane potential. Additionally, our findings confirmed that ATOX1 indirectly bound to the PPA2 promoter and increased its transcription. Notably, the restoration of ATP production in mitochondria mediated by PPA2 overexpression facilitated efficient intracellular Cu efflux, allowing rapid and precise reestablishment of intracellular Cu homeostasis under Cu stress. Collectively, Hippo-YAP signaling alleviates Cu-induced oxidative damage by restoring mitochondrial function through activation of PPA2 transcription depending on ATOX1, thereby ensuring cellular Cu efflux and enhancing antioxidant capacity.