hTBK1-c.978T>A mutation promotes the ferroptosis in NSC-34 cells via mediation of KEAP1/NRF2/p62 signaling

Background Amyotrophic lateral sclerosis (ALS) can result in the dysfunction of upper and lower motor neurons. A previous study has indicated that TBK1 mutation (hTBK1-c.978T>A) is involved in progression of ALS. However, the mechanism by which TBK1 mutation mediates the progression of ALS remains unclear. Methods NSC-34 cells with hTBK1-c.978T>A mutation (TBK1 mutation status) was used to mimic ALS in vitro. In addition, cell proliferation was detected by Ki67 staining. Gene and protein expressions in NSC-34 cells were detected by RT-qPCR and western blot, respectively. ROS and PGSK levels in NSC-34 cells were detected by flow cytometry. Results hTBK1-c.978T>A mutation significantly inhibited the proliferation of NSC-34 cells via inducing cell ferroptosis, while the effect of TBK1 mutation was notably reversed by Ferrostatin-1 or p62 siRNA. Meanwhile, hTBK1-c.978T>A mutation significantly increased the expression of KEAP1 in NSC-34 cells, while this phenomenon was partially reversed by p62 knockdown. Conclusion hTBK1-c.978T>A mutation promoted promotes the ferroptosis in NSC-34 cells via regulation of KEAP1/NRF2/p62 signaling. Thus, hTBK1-c.978T>A mutation may serve as a possible target for the treatment of ALS.