1. Academic Validation
  2. Alexidine dihydrochloride enhances the sensitivity of human hepatocellular carcinoma to disulfidptosis via ATF4-DDIT3 activation

Alexidine dihydrochloride enhances the sensitivity of human hepatocellular carcinoma to disulfidptosis via ATF4-DDIT3 activation

  • Free Radic Biol Med. 2025 Sep:237:585-599. doi: 10.1016/j.freeradbiomed.2025.06.020.
Miaomiao Li 1 Jiayi Xu 2 Ke Du 3 Gang Wang 4 Yetong Feng 5 Pengfei Liu 6
Affiliations

Affiliations

  • 1 Department of Critical Care Medicine, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China; Center for Mitochondrial Biology & Medicine, Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, China; International Joint Research Center on Cell Stress and Disease Diagnosis and Therapy, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
  • 2 International Joint Research Center on Cell Stress and Disease Diagnosis and Therapy, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
  • 3 International Joint Research Center on Cell Stress and Disease Diagnosis and Therapy, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China; Shaanxi Provincial Clinical Research Center for Hepatic & Splenic Diseases, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
  • 4 Department of Critical Care Medicine, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China; Key Laboratory of Surgical Critical Care and Life Support, Xi'an Jiaotong University, Ministry of Education of China, Xi'an, China.
  • 5 International Joint Research Center on Cell Stress and Disease Diagnosis and Therapy, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China; Core Research Laboratory, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China. Electronic address: fengyetong@xjtu.edu.cn.
  • 6 Department of Critical Care Medicine, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China; International Joint Research Center on Cell Stress and Disease Diagnosis and Therapy, National & Local Joint Engineering Research Center of Biodiagnosis and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China; Key Laboratory of Environment and Genes Related to Diseases, Xi'an Jiaotong University, Ministry of Education of China, Xi'an, China. Electronic address: liupengfei@xjtu.edu.cn.
Abstract

Disulfidptosis, an emerging regulated cell death modality characterized by pathological disulfide bond aggregation and subsequent actin cytoskeletal disintegration, presents promising therapeutic potential for apoptosis- and ferroptosis-resistant malignancies. However, the molecular initiators of Disulfidptosis and their connection with mitochondrial dysregulation remain elusive. Herein, we identified alexidine dihydrochloride (AD) as a selective Disulfidptosis enhancer in hepatocellular carcinoma (HCC), and the comprehensive multi-omics analysis and Other functional validation indicated that ATF4-DDIT3 signaling could be significantly activated by AD. Mechanistically, AD triggers severe mitochondrial stress, manifested by ultrastructural disruption, mitochondrial Reactive Oxygen Species (ROS) overproduction, and activation of the mitochondrial unfolded protein response (UPRmt). Crucially, these mitochondrial insults converge on the ATF4-DDIT3 signaling axis, which orchestrates disulfide stress amplification. This redox imbalance culminates in irreversible disulfide-mediated crosslinking of cytoskeletal proteins, the hallmark of Disulfidptosis. Additionally, knockdown of ATF4 or DDIT3 abolishes AD-enhanced Disulfidptosis, confirming their indispensable roles in this process. Totally, our findings not only characterize AD as a novel Disulfidptosis enhancer but also establish a functional crosstalk between mitochondrial dysfunction and Disulfidptosis execution, proposing a dual-target therapeutic strategy for drug resistance of HCC.

Keywords

ATF4; Alexidine dihydrochloride; DDIT3; Disulfidptosis; Hepatocellular carcinoma; Mitochondria stress.

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