The Mechanistical Study Of Histone Deacetylases On Ferroptosis

Ferroptosis is a novel form of regulated cell death found in recent years,significantly distinct from other forms of cell death in morphological,metabolic,and genetic aspects.The initiation and execution of ferroptosis are controlled by a strict and multifaceted mechanism,which is mainly characterized by iron-dependent accumulation of lipid peroxides.Increasing evidence confirms that acetylation is closely associated with the pathology of ferroptosis-related diseases.However,one essential question that remains unsolved is how HDACs regulates ferroptotic program.This study is aim to elucidate the potential involvement of HDACs in ferroptosis regulation from the aspect of acetylation,screen the key HDAC to regulate ferroptosis and clarify the underlying mechanism.The main results are as follows: 1)Firstly,we defined that HDACs negatively regulate ferroptosis by using diverse HDACs inhibitors(targeting Class Ⅰ and broad spectrum).All HDACs inhibitors enhance the accumulation of lipid ROS and facilitate cell death induced by Erastin in multiple cell lines,which could be reversed by the specific ferroptosis inhibitor Ferrostatin-1.2)We then synthesized si RNA libraries targeting the HDACs family(HDAC1-11)to screen the critical HDAC candidates that regulate ferroptosis.Notably,we identified that HDAC1 knockdown significantly facilitates Erastin-mediated ferroptosis.3)Therefore,we constructed HDAC1 knockout cell line through CRISPR/Cas9 system.HDAC1 KO promotes GSH reduction,lipid ROS accumulation,cell death and inhibition of cell proliferation under ferroptotic stress mediated by Erastin.Moreover,these effects could be reversed by Ferrostatin-1.4)Subsequently,we further explored how HDAC1 regulates ferroptosis.We found that inhibition of p53 by PFTα partially restores the ferroptosis-promoting activity of HDAC1 ablation.In addition,HDAC1 KO markedly upregulates the expression of p53 and downregulates SLC7A11,suggesting that HDAC1 negatively regulates Erastin-induced ferroptosis through p53-SLC7A11 axis.5)Intriguingly,HDAC1 KO also enhances the lipid ROS accumulation and cell death induced by another ferroptosis inducer,RSL3,which cannot be mitigated by p53 inhibition,suggesting a p53-SLC7A11 independent manner for HDAC1 modulating RSL3-mediated ferroptosis.In conclusion,we identified HDAC1 as a novel regulator of ferroptosis.Mechanistically,knockout of HDAC1 remarkably enhances GSH depletion triggered by Erastin through p53-SLC7A11 axis.Furthermore,a p53-SLC7A11 independent mechanism is underlying to HDAC1 modulating ferroptosis in response to RSL3.This study expands the biological function of HDAC1 in ferroptosis regulation.