Abstract
Doxorubicin (DOX) has toxic effects on the heart, causing cardiomyopathy and heart injury, but the underlying mechanisms of these effects require further investigation. This study investigated the role of DOX in promoting ferroptosis to induce myocardial injury. AC16 cardiomyocyte and neonatal rat ventricle cardiomyocytes were used as an in vitro model to study the molecules involved in myocardial injury using gene silencing, ectopic expression, and RNA immunoprecipitation. Messenger RNA and protein level analyses showed that DOX treatment resulted in the upregulation of methyltransferase-like 14 (METTL14), which catalyzes the m6A modification of the long non-coding RNA KCNQ1OT1, a miR-7-5p sponge. The RNA-binding protein IGF2BP1 is associated with KCNQ1OT1 to increase its stability and robustly inhibit miR-7-5p activity. Furthermore, a lack of miR-7-5p expression led to increased levels of transferrin receptor, promoting the uptake of iron and production of lipid reactive oxygen species and demonstrating that DOX-induced ferroptosis occurs in AC16 cells. Additionally, we found that miR-7-5p targets METTL14 in AC16 cells. Meanwhile, the role of METTL14/KCNQ1OT1/miR-7-5p axis in regulating ferroptosis in neonatal rat ventricle cardiomyocytes was also confirmed. Our results indicate that selectively inhibiting ferroptosis mediated by a METTL14/KCNQ1OT1/miR-7-5p positive feedback loop in cardiomyocytes could provide a new therapeutic approach to control DOX-induced cardiac injury.
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Funding
This work was funded by The Outstanding Clinical Discipline Project of Shanghai Pudong (PWYgy2018-05), Training plan of health system discipline leaders in Pudong New Area of Shanghai (PWRd2019-14), Shanghai Pudong Commission of Health and Family Planning (PW2020D-10), and Talents Training Program of the Seventh People’s Hospital, Shanghai University of Traditional Chinese Medicine (BDX2020-02).
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SZ, YM, and YZ conceived and designed the analysis. CL, FY, NJ, and JG conduct the in vitro and in vivo experiments and data analysis; HJ, CZ, and JW performed the analysis; JZ, SJ, and SZ wrote the paper. All the authors read and approved the final manuscript.
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Graphical Highlights
• METTL14 promotes DOX-induced ferroptosis in human cardiomyocytes.
• METTL14 induces the m6A modification of lncRNA KCNQ1OT1 in DOX-induced human cardiomyocytes.
• KCNQ1OT1 promotes DOX-induced ferroptosis in cardiomyocytes by sponging miR-7-5p.
• miR-7-5p inhibits DOX-induced ferroptosis in cardiomyocytes by targeting TFRC.
Shaowei Zhuang and Yan Ma both authors contributed equally to this work.
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Zhuang, S., Ma, Y., Zeng, Y. et al. METTL14 promotes doxorubicin-induced cardiomyocyte ferroptosis by regulating the KCNQ1OT1-miR-7-5p-TFRC axis. Cell Biol Toxicol 39, 1015–1035 (2023). https://doi.org/10.1007/s10565-021-09660-7
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DOI: https://doi.org/10.1007/s10565-021-09660-7