Computational and Experimental Biology Reveals Dihydroartemisinin’s Efficacy Against Steroid-Induced Osteonecrosis of the Femoral Head Adjusting Ferroptosis via CCL17-PRDX6

Yanxin Li,1– 3,* Yan Wang,1– 3,* Xiaotian Feng,1– 3,* Lulu Zhang,1– 3 Qiyu Wang,1– 3 Xuezhi Liu,1– 3 Tiancheng Ma,1– 3 Pengyuan Zhang,1– 3 Yan Du,1– 3 Mengran Qin,1– 3 Jianxiong Ma1,3 1Tianjin Hospital, Tianjin University, Tianjin, 301617, People’s Republic of China; 2Tianjin Orthopaedic Institute of Integrated Traditional Chinese and Western Medicine, Tianjin, 300050, People’s Republic of China; 3Tianjin Key Laboratory of Orthopedic Biomechanics and Medical Engineering, Tianjin, 300050, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jianxiong Ma, Tianjin Hospital, Tianjin University, Tianjin, 301617, People’s Republic of China, Email yjslwtg@126.comObjective: According to existing research findings, dihydroartemisinin effectively regulates bone metabolism balance, while ferroptosis is closely related to the occurrence of steroid-induced osteonecrosis of the femoral head. As the exact biological mechanism among the three is still unclear, Mendelian randomization, computer-aided drug design, and transcriptomics sequencing were used to explore the specific mechanism of action.Methods: The study validated the specific signaling pathways through which dihydroartemisinin may treat steroid-induced osteonecrosis of the femoral head using animal experiments and transcriptomics sequencing. Data were obtained from public databases for Mendelian randomization analysis, and a two-sample Mendelian randomization was used to determine the intermediary role of core pathway-related targets. Computer-aided drug design was employed to assess the binding affinity between dihydroartemisinin and core targets.Results: Transcriptome sequencing determined that dihydroartemisinin may treat steroid-induced osteonecrosis of the femoral head by regulating ferroptosis. We obtained 564 ferroptosis-related targets that met the analysis criteria and 1812 plasma proteins from the UK Biobank, and analyzed finngen_R11_OSTEON_DRUGS in the Finnish database as outcome. The results showed that there were two quantitative trait loci that had a causal relationship with ferroptosis targets. There were 110 protein quantitative trait loci causally associated with plasma proteins from the UK Biobank, and none of these loci had an inverse causal relationship with SONFH. Through mediation analysis, 7 mediating pathways were identified, yielding eight targets including ZP3, CCL17, APOE, C7ORF50, SPINK4, SPINK2, FTMT, and PRDX6. Computer-aided drug design revealed that CCL17 and PRDX6 exhibited the best docking effects.Conclusion: The study determined that CCL17 and PRDX6 have a significant causal relationship with SONFH. It also clarified the specific mechanism by which DHA may regulate ferroptosis to treat SONFH, which will provide a reference for the discussion of the prevention and treatment mechanisms of SONFH.Keywords: dihydroartemisinin, steroid-induced osteonecrosis of femoral head, ferroptosis, Mendelian randomization, computer-aided drug design