Many cell death pathways, including apoptosis, regulated necrosis and ferroptosis are relevant for neuronal cell death and share common mechanisms such as the formation of reactive oxygen species (ROS). However, which molecular signaling pathways contribute to related pathologies and how they are interconnected remains elusive. Here, we present the role of cofilin1 in regulating mitochondrial functions and neuronal impairment. Cofilin1 deletion in neuronal HT22 cells exerted increased mitochondrial resilience, assessed by quantification of mitochondrial ROS production, mitochondrial membrane potential and ATP levels. HT22 cells deficient for cofilin1 exhibited a profound glycolytic shift to meet their energy demand in conditions of erastin and glutamate toxicity, whereas control cells were metabolically impaired and underwent ferroptosis and oxytosis, respectively. Further, cofilin1 was confirmed as a key player in glutamate-mediated excitotoxicity in primary cortical neurons isolated from cofilin1flx/flx, CaMKII-Cre knock-out mice. Mitochondrial respiration and cell viability were significantly preserved in cofilin1-/- primary neurons under conditions of excitotoxicity. Using isolated mitochondria and recombinant cofilin1, we provide a further link to toxicity-related mitochondrial impairment mediated by oxidized cofilin1. Wildtype cofilin1 directly affected the mitochondrial membrane potential, mitochondrial ROS accumulation and mitochondrial respiration. The detrimental impact of cofilin1 on mitochondria depends on oxidation of cysteine residues at positions 139 and 147. Our findings show that the actin-regulating protein cofilin1 acts as a redox sensor in oxidative cell death pathways of ferroptosis and oxytosis, and also promotes glutamate excitotoxicity. Oxidized cofilin1 links ROS accumulation to mitochondrial demise and neuronal cell death. Protective effects by cofilin1 inhibition are particularly attributed to preserved mitochondrial integrity and function. Thus, interfering with the oxidation and pathological activation of cofilin1 may offer an effective therapeutic strategy in neurodegenerative diseases.
bioRxiv Subject Collection: Neuroscience