Abstract
Circadian rhythms disruption can be the cause of chronic diseases. External cues, including therapeutic drugs, have been shown to modulate peripheral-circadian clocks. Since anthracycline cardiotoxicity is associated with loss of mitochondrial function and metabolic remodeling, we investigated whether the energetic failure induced by sub-chronic doxorubicin (DOX) treatment in juvenile mice was associated with persistent disruption of circadian regulators. Juvenile C57BL/6J male mice were subjected to a sub-chronic DOX treatment (4 weekly injections of 5 mg/kg DOX) and several cardiac parameters, as well as circadian-gene expression and acetylation patterns, were analyzed after 6 weeks of recovery time. Complementary experiments were performed with Mouse Embryonic Fibroblasts (MEFs) and Human Embryonic Kidney 293 cells. DOX-treated juvenile mice showed cardiotoxicity markers and persistent alterations of transcriptional- and signaling cardiac circadian homeostasis. The results showed a delayed influence of DOX on gene expression, accompanied by changes in SIRT1-mediated cyclic deacetylation. The mechanism behind DOX interference with the circadian clock was further studied in vitro, in which were observed alterations of circadian-gene expression and increased BMAL1 SIRT1-mediated deacetylation. In conclusion, DOX treatment in juvenile mice resulted in disruption of oscillatory molecular mechanisms including gene expression and acetylation profiles.
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Acknowledgements
This work was funded by FEDER funds through the Operational Program for Competitiveness Factors—COMPETE and national funds by FCT—Foundation for Science and Technology under research Grant PTDC/DTP-FTO/2433/2014 (POCI-01-0145-FEDER-016659), PTDC/BTM-SAL/29297/2017 (POCI-01-0145-FEDER-029297), and UID/NEU/04539/2019. Supported also by QREN project 4832 with reference CENTRO- 07-ST24-FEDER-002008 financed through FEDER. LF was supported by an FCT PhD-fellowship (SFRH/BD/ 52429/2013), and TC-O was supported by Grant POCI-01-0145-FEDER-029297.
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LF and PC designed the experiments. LF conducted the experiments and collected the data with early technical assistance from MC. Raw sequencing data was analyzed by HF and CE. LF analyzed the data, wrote the manuscript, and prepared all figures. TO finalized the manuscript. LF, PO, TO, MC, and PC reviewed the manuscript.
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Ferreira, L.L., Cervantes, M., Froufe, H.J.C. et al. Doxorubicin persistently rewires cardiac circadian homeostasis in mice. Arch Toxicol 94, 257–271 (2020). https://doi.org/10.1007/s00204-019-02626-z
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DOI: https://doi.org/10.1007/s00204-019-02626-z