Mito-tempol and dexrazoxane exhibit cardioprotective and chemotherapeutic effects through specific protein oxidation and autophagy in a syngeneic breast tumor preclinical model.
Several front-line chemotherapeutics cause mitochondria-derived, oxidative stress-mediated cardiotoxicity. Iron chelators and other antioxidants have not completely succeeded in mitigating this effect. One hindrance to the development of cardioprotectants is the lack of physiologically-relevant animal models to simultaneously study antitumor activity and cardioprotection. Therefore, we optimized a syngeneic rat model and examined the mechanisms by which oxidative stress affects outcome. Immune-competent spontaneously hypertensive rats (SHRs) were implanted with passaged, SHR-derived, breast tumor cell line, SST-2. Tumor growth and cytokine responses (IL-1A, MCP-1, TNF-α) were observed for two weeks post-implantation. To demonstrate the utility of the SHR/SST-2 model for monitoring both anticancer efficacy and cardiotoxicity, we tested cardiotoxic doxorubicin alone and in combination with an established cardioprotectant, dexrazoxane, or a nitroxide conjugated to a triphenylphosphonium cation, Mito-Tempol (4) [Mito-T (4)]. As predicted, tumor reduction and cardiomyopathy were demonstrated by doxorubicin. We confirmed mitochondrial accumulation of Mito-T (4) in tumor and cardiac tissue. Dexrazoxane and Mito-T (4) ameliorated doxorubicin-induced cardiomyopathy without altering the antitumor activity. Both agents increased the pro-survival autophagy marker LC3-II and decreased the apoptosis marker caspase-3 in the heart, independently and in combination with doxorubicin. Histopathology and transmission electron microscopy demonstrated apoptosis, autophagy, and necrosis corresponding to cytotoxicity in the tumor and cardioprotection in the heart. Changes in serum levels of 8-oxo-dG-modified DNA and total protein carbonylation corresponded to cardioprotective activity. Finally, 2D-electrophoresis/mass spectrometry identified specific serum proteins oxidized under cardiotoxic conditions. Our results demonstrate the utility of the SHR/SST-2 model and the potential of mitochondrially-directed agents to mitigate oxidative stress-induced cardiotoxicity. Our findings also emphasize the novel role of specific protein oxidation markers and autophagic mechanisms for cardioprotection.
Division of Therapeutic Proteins, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA.
Jennifer S
JS
Dickey
Yanira
Y
Gonzalez
Baikuntha
B
Aryal
Steven
S
Mog
Asako J
AJ
Nakamura
Christophe E
CE
Redon
Ulrich
U
Baxa
Elliot
E
Rosen
Gang
G
Cheng
Jacek
J
Zielonka
Palak
P
Parekh
Karen P
KP
Mason
Joy
J
Joseph
Balaraman
B
Kalyanaraman
William
W
Bonner
Eugene
E
Herman
Emily
E
Shacter
V Ashutosh
VA
Rao
D000818:Animals; D000975:Antioxidants / Q000627:therapeutic use; D001343:Autophagy / Q000187:drug effects*; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism*; D045744:Cell Line, Tumor; D064730:Dexrazoxane / Q000627:therapeutic use*; D004195:Disease Models, Animal; D005260:Female; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000627:therapeutic use*; D010084:Oxidation-Reduction / Q000187:drug effects; D010880:Piperidines / Q000627:therapeutic use*; D050050:Protein Carbonylation / Q000187:drug effects; D051381:Rats; D011918:Rats, Inbred SHR
D000975:Antioxidants; D064730:Dexrazoxane; D008869:Microtubule-Associated Proteins; D009943:Organophosphorus Compounds; D010880:Piperidines; C526109:Anxa3 protein, rat; C555916:MitoTEMPO