Development of doxorubicin-induced chronic cardiotoxicity in the B6C3F1 mouse model
Highlights
► 24 mg/kg was a cumulative cardiotoxic dose of doxorubicin in male B6C3F1 mice. ► Doxorubicin-induced hematological toxicity was in association with splenomegaly. ► Doxorubicin induced severe testicular toxicity in B6C3F1 male mice.
Introduction
Doxorubicin (DOX) is one of the most potent anthracycline antibiotics widely used to treat several types of solid tumors and hematologic malignancies (Ahmad et al., 2012, Batty et al., 2012, Carr, 2004, Honda et al., 2010, Masci et al., 2004, Stavridi and Palmieri, 2008). However, one of the serious complications associated with the clinical use of the drug is the risk of developing dose-related cumulative and irreversible cardiomyopathy, which can lead to life-threatening congestive heart failure (reviewed in Wouters et al., 2005). Among several biochemical markers of cardiac tissue damage, troponins are routinely utilized as the most sensitive and specific clinical biomarkers of myocardial injury (Lipshultz et al., 2004, Lipshultz et al., 2011). Several preclinical studies also have demonstrated the usefulness of serum levels of cardiac troponin T in determining the extent of myocardial damage caused by DOX (Bertinchant et al., 2003, Herman et al., 1998, Herman et al., 1999, Herman et al., 2001). However, these proteins are released in serum only after cardiac tissue damage has occurred. To control or prevent damage to the heart tissue during DOX treatment, it is crucial to identify biomarkers that would predict early stages of cardiac events before irreversible damage has occurred.
Doxorubicin has been extensively investigated for toxic effects of cumulative doses in the heart in various animal models. These include the rabbit (Herman and Ferrans, 1983, Jaenke, 1976, Solcia et al., 1981, Van Vleet and Ferrans, 1980), dog (Astra et al., 2003, Herman and Ferrans, 1983, Solcia et al., 1981, Tomlinson et al., 1986), pig (Van Vleet et al., 1979), monkey (Denine and Schmidt, 1975, Sieber et al., 1980), and the rat (Berthiaume and Wallace, 2007, Herman et al., 1985, Lebrecht et al., 2003, Solcia et al., 1981, Thompson et al., 2010, Zhou et al., 2001). Several studies have utilized mouse models in the evaluation of DOX-induced cardiac toxicity. However, these studies have primarily examined molecular events of cardiac toxicity at acute high doses instead of the chronic doses of DOX used in the clinical situation (Bjelogrlic et al., 2007, Doroshow et al., 1981, Matsumura et al., 1994, Olson et al., 2003, Papadopoulou et al., 1999, Rosenoff et al., 1975, Shuai et al., 2011, Tan et al., 2011). In addition, DOX was administered to the mice intraperitoneally, whereas DOX is administered intravenously in cancer patients. As a result, information concerning cardiotoxicity caused by chronic DOX exposure in mice is inadequate. One of the limitations of using the mouse in chronic toxicity studies is the difficulty of administering multiple intravenous injections of DOX over an extended period of time (Herman and Ferrans, 1998). This may, in part, be due to the ability of the drug to cause localized tissue damage.
Development of a mouse model of DOX-induced cardiotoxicity using the clinically relevant treatment strategy of smaller doses over a longer period of time will provide important insights into the early molecular events during drug exposure. This further will allow identification of early molecular markers of DOX-induced cardiac tissue injury that leads to a release of troponins. Genomic and genetic tools, such as a well-annotated complete genome sequence and transgenic models, are uniquely available for the mouse system. Application of these powerful genetic methods in the mouse can help validate the identified biomarkers for drug-induced cardiac tissue damage. In addition, comparison of functional and toxicological data from extensively investigated chronically exposed animal models to the findings in this newly developed chronic cardiotoxicity mouse model could advance our mechanistic understanding of DOX-mediated cardiotoxicity across the species, which may lead to the design of new, improved predictive biomarkers of cardiotoxicity. Translation of such biomarkers to the clinic may help in the prediction or early diagnosis of adverse cardiac events and also in designing novel treatment strategies to prevent or alleviate unwanted adverse events during DOX therapy.
Section snippets
Animal husbandry
Male B6C3F1 mice were provided as weanlings by the breeding colony at the National Center for Toxicological Research (NCTR). Animals were raised in a pathogen-free environment at the NCTR and treated according to the Institutional Animal Care and Use Committee (IACUC) guidelines. At 6 weeks of age, mice were housed individually in standard polycarbonate cages with hardwood chip bedding and were maintained at 23 °C with a relative humidity of 50%. The animals were conditioned to a 12/12-h
Animal survival, changes in body weights, and daily food consumption
No mortality was observed in any of the treatment groups. Both DOX- and saline-treated mice showed gains in their body weights during 15 weeks on the study. The mean increase in body weight in DOX-treated mice was 3.5 g (14%) whereas the mean body weight gain was 13.9 g (54%) in saline-treated mice at the end of the study. Average body weight at the end of the study was 29.5 ± 0.7 g in DOX-treated mice and 39.8 ± 0.5 g in saline-treated mice. The differences in body weights between DOX-treated and
Discussion
A dose-dependent cumulative cardiac toxicity of DOX remains a major clinical concern and has led to the development of several animal models of chronic cardiotoxicity to investigate the molecular basis of progressive cardiac dysfunction and various cardioprotective strategies (Bertazzoli et al., 1979, Berthiaume and Wallace, 2007, Denine and Schmidt, 1975, Gralla et al., 1979, Herman and Ferrans, 1983, Herman and Ferrans, 1998, Herman et al., 1983, Herman et al., 1985, Herman et al., 2000, Van
Conflict of interest statement
The authors declare that there are no conflicts of interest.
Disclaimer
The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the FDA.
Acknowledgments
The authors greatly appreciate Kim Knight and Jennifer Garrison for their excellent technical skills in administering doxorubicin intravenously in mice, and also, Andy Matson, for his expertise in preparations of doxorubicin dose solutions to the greatest accuracy. The authors also express their gratitude to Drs. Margaret Miller and Mugimane Manjanatha for the critical review of the manuscript.
References (94)
- et al.
Primary testicular lymphoma
Clin. Oncol.
(2012) - et al.
Evaluation of cardiac troponin I and T levels as markers of myocardial damage in doxorubicin-induced cardiomyopathy rats, and their relationship with echocardiographic and histological findings
Clin. Chim. Acta
(2003) - et al.
Anthracycline cardiotoxicity in transgenic mice overexpressing SR Ca2 + − ATPase
Biochem. Biophys. Res. Commun.
(2003) Hepatocellular carcinoma: current management and future trends
Gastroenterology
(2004)- et al.
Transcriptional profiling for understanding the basis of mitochondrial involvement in disease and toxicity using the mitochondria-specific MitoChip
Mutat. Res. Fundam. Mol. Mech. Mutagen.
(2007) - et al.
Development of mitochondria-specific mouse oligonucleotide microarray and validation of data by real-time PCR
Mitochondrion
(2007) - et al.
Nucleoside reverse transcriptase inhibitors (NRTIs)-induced expression profile of mitochondria-related genes in the mouse liver
Mitochondrion
(2008) - et al.
The dosing schedule dependent toxicities of adriamycin in beagle dogs and rhesus monkey
Toxicology
(1979) - et al.
Comparison of the severity of the chronic cardiotoxicity produced by doxorubicin in normotensive and hypertensive rats
Toxicol. Appl. Pharmacol.
(1985) - et al.
Effect of (+)-usnic acid on mitochondrial functions as measured by mitochondria-specific oligonucleotide microarray in liver of B6C3F1 mice
Mitochondrion
(2009)
Testicular cytotoxicity of intravenous doxorubicin in rats
J. Urol.
Age-related acute adriamycin cardiotoxicity in mice
J. Mol. Cell. Cardiol.
Structural and functional impairment of mitochondria in adriamycin-induced cardiomyopathy in mice: suppression of cytochrome c oxidase II gene expression
Biochem. Pharmacol.
Cardiotoxicity of cytotoxic drugs
Cancer Treat. Rev.
Global gene expression profiles of MT knockout and wild-type mice in the condition of doxorubicin-induced cardiomyopathy
Toxicol. Lett.
A randomized placebo-controlled trial of recombinant human interleukin-11 in cancer patients with severe thrombocytopenia due to chemotherapy
Blood
Berberine attenuates doxorubicin-induced cardiotoxicity in mice
J. Int. Med. Res.
Doxorubicin-induced persistent oxidative stress to cardiac myocytes
Toxicol. Lett.
Effects of actinomycin D in vivo on murine erythroid stem cells
Blood
Doxorubicin-induced canine CHF: advantages and disadvantages
J. Card. Surg.
Anthracycline antibiotics induce acute renal tubular toxicity in children with cancer
Pathol. Oncol. Res.
Doxorubicin, bleomycin, vinblastine and dacarbazine chemotherapy with interferon for advanced stage classic Hodgkin lymphoma: a 10-year follow-up study
Leuk. Lymphoma
Quantitative experimental evaluation of adriamycin cardiotoxicity in the mouse
Cancer Treat. Rep.
Persistent alterations to the gene expression profile of the heart subsequent to chronic doxorubicin treatment
Cardiovasc. Toxicol.
In vivo cardiac electrophysiology studies in the mouse
Circulation
Anthracycline cardiomyopathy monitored by morphologic changes
Cancer Treat. Rep.
Effects of dexrazoxane and amifostine on evolution of doxorubicin cardiomyopathy in vivo
Exp. Biol. Med.
Prevention and treatment of organ toxicity during high-dose chemotherapy: an overview
Anticancer Drugs
Systemic treatment-induced gastrointestinal toxicity: incidence, clinical presentation and management
Ann. Gastroenterol.
Adjuvant sequential dose-dense doxorubicin, paclitaxel, and cyclophosphamide (ATC) for high-risk breast cancer is feasible in the community setting
Cancer J. Sci. Am.
Doxorubicin: the good, the bad and the ugly effect
Curr. Med. Chem.
Guidance for Industry. Estimating the maximum safe starting dose in initial clinical trials for therapeutics in adult healthy volunteers
Antitumor activity and toxicological properties of doxorubicin conjugated to (alpha),(beta)-poly[(2-hydroxyethyl)-L-aspartamide] administered intraperitoneally in mice
Anticancer Drugs
Measurement of cardiac troponins
Ann. Clin. Biochem.
Lost in transgenesis: a user's guide for genetically manipulating the mouse in cardiac research
Circ. Res.
Adriamycin induced myopathies in the rhesus monkey with emphasis on cardiomyopathy
Toxicol. Appl. Pharmacol.
Cardiac-specific inducible and conditional gene targeting in mice
Circ. Res.
Prevention of doxorubicin cardiac toxicity in the mouse by N-acetylcysteine
J. Clin. Invest.
Ghrelin attenuates gastrointestinal epithelial damage induced by doxorubicin
World J. Gastroenterol.
Importance of metabolic changes induced by chemotherapy on prognosis of early-stage breast cancer patients: a review of potential mechanisms
Obes. Rev.
ICRF-187: reduction of chronic daunorubicin and doxorubicin cardiotoxicity in rabbits, beagle dogs and miniature pigs
Drugs Exp. Clin. Res.
Preclinical animal models of cardiac protection from anthracycline-induced cardiotoxicity
Semin. Oncol.
Prevention of chronic doxorubicin cardiotoxicity in beagles by liposomal encapsulation
Cancer Res.
Use of cardiac troponin T levels as an indicator of doxorubicin-induced cardiotoxicity
Cancer Res.
Correlation between serum levels of cardiac troponin-T and the severity of the chronic cardiomyopathy induced by doxorubicin
J. Clin. Oncol.
Comparison of the protective effects of amifostine and dexrazoxane against the toxicity of doxorubicin in spontaneously hypertensive rats
Cancer Chemother. Pharmacol.
The use of serum levels of cardiac troponin T to compare the protective activity of dexrazoxane against doxorubicinand mitoxantrone-induced cardiotoxicity
Cancer Chemother. Pharmacol.
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