NCOA4-dependent ferritinophagy drives perfluorooctanoic acid-triggered renal ferroptosis in chicken embryos

Perfluorooctanoic acid (PFOA), a persistent organic pollutant, is widely distributed in the environment, leading to substantial bioaccumulation and posing significant public health concerns. The kidney is the primary target organ for PFOA, but a recognized mechanism remains unclear. The chicken embryo has been widely used as a maternal-independent model in toxicity studies. However, the toxic effects of PFOA in chicken embryo kidneys remain limited. In the present study, chicken embryos were injected with PFOA of different concentrations through the air cell to establish an in vivo model, and establish an in vitro model of chicken embryo kidney (CEK) cells for PFOA exposure. The results showed that PFOA promoted nephrotoxicity and induced ferroptosis in chicken embryo kidneys. Meanwhile, ferroptosis occurred in PFOA-treated CEK cells and was alleviated by ferroptosis inhibitor Ferrostatin-1 (Fer-1), Deferoxamine (DFO), and autophagy inhibitor Chloroquine (CQ). Further results suggested that PFOA activated nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy in chicken embryo kidneys and CEK cells. Mechanistically, PFOA diminished NCOA4 ubiquitination by reducing the expression of HECT and RLD domain containing E3 ubiquitin protein ligase 2 (HERC2), leading to increased NCOA4 expression and activated ferritinophagy. These findings indicated that PFOA induced ferroptosis via HERC2-regulated NCOA4 ubiquitination-dependent degradation in chicken embryo kidneys.