Proteotoxic stress-induced autophagy is regulated by the NRF2 pathway via extracellular vesicles

Protein homeostasis involves a number of overlapping mechanisms, including the autophagy program, that can lead to the resolution of protein damage. We aimed in this study to examine mechanisms of autophagy in the proteotoxic stress response. We found that such stress results in a rapid elevation in the rate of autophagy in mammalian cells. Induction of this process occurred coincidentally with the increased release of extracellular vesicles (EVs) into the extracellular microenvironment. We next found that purified EVs that had been released from stressed cells were capable of directly increasing autophagic flux in recipient cells. The EVs contained a range of cargo proteins, including HSP70, BAG3, and activated transcription factor phospho-NRF2 (pNRF2). NRF2 regulates the activation of both the oxidative stress response and autophagy genes. Both heat shock and exposure of cells to proteotoxic stress-induced EVs increased the intracellular levels of pNRF2 in cells. Heat shock-induced proteotoxicity also led to increases in the levels of proteins in the oxidative stress response, including HO-1 and NQO1, as well as the key autophagy proteins LC3, ATG5, and ATG7, known to be regulated by NRF2. Increases in these autophagy proteins were dependent on the expression of NRF2 and were ablated by NRF2 knockdown.