Abstract
The use of anthracycline chemotherapeutic drugs is restricted owing to potentially fatal cardiotoxic side effects. It has been hypothesized that anthracycline metabolites have a primary role in this cardiac dysfunction; however, information on the molecular interactions of these compounds in the heart is scarce. Here we provide novel evidence that doxorubicin and its metabolite, doxorubicinol, bind to the cardiac ryanodine receptor (RyR2) and to the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA2A) and deleteriously alter their activity. Both drugs (0.01 μM–2.5 μM) activated single RyR2 channels, and this was reversed by drug washout. Both drugs caused a secondary inhibition of RyR2 activity that was not reversed by drug washout. Preincubation with the reducing agent dithiothreitol (DTT, 1 mM) prevented drug-induced inhibition of channel activity. Doxorubicin and doxorubicinol reduced the abundance of thiol groups on RyR2, further indicating that oxidation reactions may be involved in the actions of the compounds. Ca2+ uptake into sarcoplasmic reticulum vesicles by SERCA2A was inhibited by doxorubicinol, but not doxorubicin. Unexpectedly, in the presence of DTT, doxorubicinol enhanced the rate of Ca2+ uptake by SERCA2A. Together the evidence provided here shows that doxorubicin and doxorubicinol interact with RyR2 and SERCA2A in similar ways, but that the metabolite acts with greater efficacy than the parent compound. Both compounds modify RyR2 and SERCA2A activity by binding to the proteins and also act via thiol oxidation to disrupt SR Ca2+ handling. These actions would have severe consequences on cardiomyocyte function and contribute to clinical symptoms of acute anthracycline cardiotoxicity.
Footnotes
- Received June 4, 2014.
- Accepted August 8, 2014.
↵1 Current affiliation: Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas.
↵2 Current affiliation: Discipline of Biomedical Sciences, Centre for Research in Therapeutic Solutions, Faculty of Education, Science, Technology and Maths, University of Canberra, Canberra, ACT, Australia.
This work was supported by the National Health and Medical Research Council Project grant [APP471400] to N.A.B, a National Health and Medical Research Council Project grant [APP1021342] to A.F.D., and N.A.B, and a National Health and Medical Research Council Career Development Award [APP1003985] to N.A.B.
- Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics
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