A CRISPR-Cas9 library screening identifies CARM1 as a critical inhibitor of ferroptosis in hepatocellular carcinoma cells

Yiming Cheng; Xiaochen Wang; Shuyu Huang; Liang Zhang; Bei Lan; Xuanyuan Li; Hao Chen; Zhenfeng Liu; Yijie Su; Lishan Xi; Shengyun Feng; Yanxuan Guo; Jun Zhou; Yingmei Wang; Chenghao Xuan

doi:10.1016/j.omtn.2023.102063

A CRISPR-Cas9 library screening identifies CARM1 as a critical inhibitor of ferroptosis in hepatocellular carcinoma cells

Mol Ther Nucleic Acids. 2023 Oct 20:34:102063. doi: 10.1016/j.omtn.2023.102063. eCollection 2023 Dec 12.

Authors

Yiming Cheng¹, Xiaochen Wang¹, Shuyu Huang¹, Liang Zhang², Bei Lan¹, Xuanyuan Li¹, Hao Chen¹, Zhenfeng Liu¹, Yijie Su¹, Lishan Xi¹, Shengyun Feng¹, Yanxuan Guo¹, Jun Zhou³, Yingmei Wang⁴, Chenghao Xuan¹

Affiliations

¹ Tianjin Key Laboratory of Female Reproductive Health and Eugenetics, Tianjin Medical University General Hospital, The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, Department of Biochemistry and Molecular Biology, Tianjin Medical University, Tianjin 300070, China.
² Research Center of Translational Medicine, Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong 250013, China.
³ Institute of Biomedical Sciences, College of Life Sciences, Shandong Normal University, Jinan, Shandong 250014, China.
⁴ Department of Gynecology and Obstetrics, Tianjin Medical University General Hospital, Tianjin 300052, China.

Abstract

Ferroptosis is an iron-catalyzed form of regulated cell death that results from the accumulation of lipid peroxidation products and reactive oxygen species to a lethal content. However, the transcriptional regulation of ferroptosis is not well understood. Sorafenib, a standard drug for hepatocellular carcinoma (HCC), induces ferroptosis in HCC cells. In this study, we conducted a CRISPR-Cas9 library screening targeting epigenetic factors and identified coactivator-associated arginine methyltransferase 1 (CARM1) as a critical inhibitor of ferroptosis. CARM1 depletion intensified Sorafenib-induced ferroptosis, resulting in decreased cell viability, reduced cellular glutathione level, increased lipid peroxidation, and altered mitochondrial crista structure. Additionally, we investigated a CARM1 inhibitor (CARM1i) as a potential ferroptosis inducer. Combining the CARM1i with Sorafenib enhanced the induction of ferroptosis. Notably, both CARM1 knockdown and CARM1i showed cooperative effects with Sorafenib in inhibiting HCC growth in mice. The underlying mechanism involves CARM1-catalyzed H3R26me2a on the promoter of glutathione peroxidase 4, leading to its transcriptional activation and subsequent ferroptosis inhibition. Furthermore, Sorafenib treatment induced the transcription of CARM1 through the MDM2-p53 axis. In summary, our findings establish CARM1 as a critical ferroptosis inhibitor and highlight the potential of CARM1is as novel ferroptosis inducers, providing promising therapeutic strategies for HCC treatment.

Keywords: CARM1; CRISPR-Cas9; GPX4; MT: Bioinformatics; Sorafenib; ferroptosis.