Chronic anthracycline cardiotoxicity is a serious clinical issue with well characterized functional and histopathological hallmarks. However, molecular determinants of the toxic damage and associated myocardial remodeling remain to be established. Furthermore, details on the different propensity of the left and right ventricle (LV and RV, respectively) to the cardiotoxicity development are unknown. Hence, the aim of the investigation was to study molecular changes associated with remodeling of the LV and RV in chronic anthracycline cardiotoxicity and post-treatment follow up. The cardiotoxicity was induced in rabbits with daunorubicin (3 mg/kg/week for 10 weeks) and animals were sacrificed either at the end of the treatment or after an additional 10 weeks. Daunorubicin induced severe and irreversible cardiotoxicity associated with LV dysfunction and typical morphological alterations, whereas the myocardium of the RV showed only mild changes. Both ventricles also showed different expression of ANP after daunorubicin treatment. Daunorubicin impaired the expression of several sarcomeric proteins in the LV, which was not the case of the RV. In particular, a significant drop was found in titin and thick filament proteins at both mRNA and protein level and this might be connected with persistent LV down-regulation of GATA-4. In addition, the LV was more affected by treatment-induced perturbations in calcium handling proteins. LV cardiomyocytes showed marked up-regulation of desmin after the treatment and vimentin was mainly induced in LV fibroblasts, whereas only weaker changes were observed in the RV. Remodeling of extracellular matrix was almost exclusively found in the LV with particular induction of collagen I and IV. Hence, the present study describes profound molecular remodeling of myocytes, non-myocyte cells and extracellular matrix in response to chronic anthracycline treatment with marked asymmetry between LV and RV.

Department of Biochemical Sciences Faculty of Pharmacy in Hradec Králové Charles University Prague Hradec Králové Czech Republic

Department of Histology and Embryology Faculty of Medicine in Hradec Králové Charles University Prague Hradec Králové Czech Republic

Department of Pharmacology Faculty of Medicine in Hradec Králové Charles University Prague Hradec Králové Czech Republic

Department of Physiology Faculty of Medicine in Hradec Králové Charles University Prague Hradec Králové Czech Republic

Institute of Molecular Pathology Faculty of Military Health Sciences University of Defence Hradec Králové Czech Republic; Department of Biochemical Sciences Faculty of Pharmacy in Hradec Králové Charles University Prague Hradec Králové Czech Republic

Zobrazit více v PubMed

Jones RL, Swanton C, Ewer MS (2006) Anthracycline cardiotoxicity. Expert Opin Drug Saf 5: 791–809. PubMed

Menna P, Paz OG, Chello M, Covino E, Salvatorelli E, et al. (2012) Anthracycline cardiotoxicity. Expert Opin Drug Saf 11 Suppl 1S21–36. PubMed

Von Hoff DD, Layard MW, Basa P, Davis HL Jr, Von Hoff AL, et al. (1979) Risk factors for doxorubicin-induced congestive heart failure. Ann Intern Med 91: 710–717. PubMed

Van Vleet JF, Ferrans VJ (1980) Clinical and pathologic features of chronic adriamycin toxicosis in rabbits. Am J Vet Res 41: 1462–1469. PubMed

Billingham ME, Mason JW, Bristow MR, Daniels JR (1978) Anthracycline cardiomyopathy monitored by morphologic changes. Cancer Treat Rep 62: 865–872. PubMed

Berry GJ, Jorden M (2005) Pathology of radiation and anthracycline cardiotoxicity. Pediatr Blood Cancer 44: 630–637. PubMed

Van Vleet JF, Ferrans VJ, Weirich WE (1980) Cardiac disease induced by chronic adriamycin administration in dogs and an evaluation of vitamin E and selenium as cardioprotectants. Am J Pathol 99: 13–42. PubMed PMC

Rhoden W, Hasleton P, Brooks N (1993) Anthracyclines and the Heart. Br Heart J 70: 499–502. PubMed PMC

Herman EH, Ferrans VJ (1998) Preclinical animal models of cardiac protection from anthracycline-induced cardiotoxicity. Semin Oncol 25: 15–21. PubMed

Sterba M, Popelova O, Vavrova A, Jirkovsky E, Kovarikova P, et al. (2013) Oxidative stress, redox signaling, and metal chelation in anthracycline cardiotoxicity and pharmacological cardioprotection. Antioxid Redox Signal 18: 899–929. PubMed PMC

Vejpongsa P, Yeh ET (2013) Topoisomerase 2beta: A Promising Molecular Target for Primary Prevention of Anthracycline-induced Cardiotoxicity. Clin Pharmacol Ther doi: 10.1038/clpt.2013.201 PubMed DOI

Sawyer DB, Peng X, Chen B, Pentassuglia L, Lim CC (2010) Mechanisms of anthracycline cardiac injury: can we identify strategies for cardioprotection? Prog Cardiovasc Dis 53: 105–113. PubMed PMC

Boucek RJ Jr, Miracle A, Anderson M, Engelman R, Atkinson J, et al. (1999) Persistent effects of doxorubicin on cardiac gene expression. J Mol Cell Cardiol 31: 1435–1446. PubMed

Ito H, Miller SC, Billingham ME, Akimoto H, Torti SV, et al. (1990) Doxorubicin selectively inhibits muscle gene expression in cardiac muscle cells in vivo and in vitro. Proc Natl Acad Sci U S A 87: 4275–4279. PubMed PMC

Aries A, Paradis P, Lefebvre C, Schwartz RJ, Nemer M (2004) Essential role of GATA-4 in cell survival and drug-induced cardiotoxicity. Proc Natl Acad Sci U S A 101: 6975–6980. PubMed PMC

Jeyaseelan R, Poizat C, Baker RK, Abdishoo S, Isterabadi LB, et al. (1997) A novel cardiac-restricted target for doxorubicin - CARP, a nuclear modulator of gene expression in cardiac progenitor cells and cardiomyocytes. J Biol Chem 272: 22800–22808. PubMed

Kobayashi S, Volden P, Timm D, Mao K, Xu XM, et al. (2010) Transcription Factor GATA4 Inhibits Doxorubicin-induced Autophagy and Cardiomyocyte Death. J Biol Chem 285: 793–804. PubMed PMC

Chen B, Zhong L, Roush SF, Pentassuglia L, Peng XY, et al... (2012) Disruption of a GATA4/Ankrd1 Signaling Axis in Cardiomyocytes Leads to Sarcomere Disarray: Implications for Anthracycline Cardiomyopathy. Plos One 7. PubMed PMC

Lim CC, Zuppinger C, Guo X, Kuster GM, Helmes M, et al. (2004) Anthracyclines induce calpain-dependent titin proteolysis and necrosis in cardiomyocytes. J Biol Chem 279: 8290–8299. PubMed

Li YF, Wang XJ (2011) The role of the proteasome in heart disease. Biochim Biophys Acta 1809: 141–149. PubMed PMC

Institute of Laboratory Animal Research CoLS, National Research Council (1996) Guide for the Care and Use of Laboratory Animals. Washington: The National Academies Press.

Popelova O, Sterba M, Haskova P, Simunek T, Hroch M, et al. (2009) Dexrazoxane-afforded protection against chronic anthracycline cardiotoxicity in vivo: effective rescue of cardiomyocytes from apoptotic cell death. Br J Cancer 101: 792–802. PubMed PMC

Simunek T, Klimtova I, Kaplanova J, Mazurova Y, Adamcova M, et al. (2004) Rabbit model for in vivo study of anthracycline-induced heart failure and for the evaluation of protective agents. Eur J Heart Fail 6: 377–387. PubMed

Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29: e45. PubMed PMC

Tatsumi R, Hattori A (1995) Detection of giant myofibrillar proteins connectin and nebulin by electrophoresis in 2% polyacrylamide slab gels strengthened with agarose. Anal Biochem 224: 28–31. PubMed

Reddy GK, Enwemeka CS (1996) A simplified method for the analysis of hydroxyproline in biological tissues. Clin Biochem 29: 225–229. PubMed

Ewer MS, Benjamin RS (2006) Doxorubicin Cardiotoxicity: Clinical Apect, Recognition, Monitoring, Treatment, and Prevention. In: Ewer MS, Yeh E, editors. Cancer and the Heart. Hamilton: BC Decker. 9–32.

Sterba M, Popelova O, Lenco J, Fucikova A, Brcakova E, et al. (2011) Proteomic insights into chronic anthracycline cardiotoxicity. J Mol Cell Cardiol 50: 849–862. PubMed

Moretti A, Weig HJ, Ott T, Seyfarth M, Holthoff HP, et al. (2002) Essential myosin light chain as a target for caspase-3 in failing myocardium. Proc Natl Acad Sci U S A 99: 11860–11865. PubMed PMC

Hernandez OM, Jones M, Guzman G, Szczesna-Cordary D (2007) Myosin essential light chain in health and disease. Am J Physiol Heart Circ Physiol 292: H1643–H1654. PubMed

De Beer EL, Bottone AE, van der Velden J, Voest EE (2000) Doxorubicin impairs crossbridge turnover kinetics in skinned cardiac trabeculae after acute and chronic treatment. Mol Pharmacol 57: 1152–1157. PubMed

Gupta MP (2007) Factors controlling cardiac myosin-isoform shift during hypertrophy and heart failure. J Mol Cell Cardiol 43: 388–403. PubMed PMC

Arai M, Tomaru K, Takizawa T, Sekiguchi K, Yokoyama T, et al. (1998) Sarcoplasmic reticulum genes are selectively down-regulated in cardiomyopathy produced by doxorubicin in rabbits. J Mol Cell Cardiol 30: 243–254. PubMed

Makarenko I, Opitz CA, Leake MC, Neagoe C, Kulke M, et al. (2004) Passive stiffness changes caused by upregulation of compliant titin isoforms in human dilated cardiomyopathy hearts. Circ Res 95: 708–716. PubMed

Miller MK, Bang ML, Witt CC, Labeit D, Trombitas C, et al. (2003) The muscle ankyrin repeat proteins: CARP, ankrd2/Arpp and DARP as a family of titin filament-based stress response molecules. J Mol Biol 333: 951–964. PubMed

Torrado M, Lopez E, Centeno A, Castro-Beiras A, Mikhailov AT (2004) Left-right asymmetric ventricular expression of CARP in the piglet heart: regional response to experimental heart failure. Eur J Heart Fail 6: 161–172. PubMed

Kim Y, Ma AG, Kitta K, Fitch SN, Ikeda T, et al. (2003) Anthracycline-induced suppression of GATA-4 transcription factor: implication in the regulation of cardiac myocyte apoptosis. Mol Pharmacol 63: 368–377. PubMed

Li L, Takemura G, Li Y, Miyata S, Esaki M, et al. (2006) Preventive effect of erythropoietin on cardiac dysfunction in doxorubicin-induced cardiomyopathy. Circulation 113: 535–543. PubMed

Palumbo A, Gay F, Bringhen S, Falcone A, Pescosta N, et al. (2008) Bortezomib, doxorubicin and dexamethasone in advanced multiple myeloma. Ann Oncol 19: 1160–1165. PubMed

Eghbali M, Czaja MJ, Zeydel M, Weiner FR, Zern MA, et al. (1988) Collagen chain mRNAs in isolated heart cells from young and adult rats. J Mol Cell Cardiol 20: 267–276. PubMed

Chapman D, Weber KT, Eghbali M (1990) Regulation of fibrillar collagen types I and III and basement membrane type IV collagen gene expression in pressure overloaded rat myocardium. Circ Res 67: 787–794. PubMed

Villarreal FJ, Dillmann WH (1992) Cardiac hypertrophy-induced changes in mRNA levels for TGF-beta 1, fibronectin, and collagen. Am J Physiol 262: H1861–1866. PubMed

Chen MM, Lam A, Abraham JA, Schreiner GF, Joly AH (2000) CTGF expression is induced by TGF- beta in cardiac fibroblasts and cardiac myocytes: a potential role in heart fibrosis. J Mol Cell Cardiol 32: 1805–1819. PubMed

Yi X, Bekeredjian R, DeFilippis NJ, Siddiquee Z, Fernandez E, et al. (2006) Transcriptional analysis of doxorubicin-induced cardiotoxicity. Am J Physiol Heart Circ Physiol 290: H1098–1102. PubMed

Luchner A, Muders F, Dietl O, Friedrich E, Blumberg F, et al. (2001) Differential expression of cardiac ANP and BNP in a rabbit model of progressive left ventricular dysfunction. Cardiovasc Res 51: 601–607. PubMed

Simunek T, Sterba M, Holeckova M, Kaplanova J, Klimtova I, et al. (2005) Myocardial content of selected elements in experimental anthracycline-induced cardiomyopathy in rabbits. Biometals 18: 163–169. PubMed

Goldstein JA (2002) Pathophysiology and management of right heart ischemia. J Am Coll Cardiol 40: 841–853. PubMed

Walker LA, Buttrick PM (2009) The right ventricle: biologic insights and response to disease. Curr Cardiol Rev 5: 22–28. PubMed PMC

Cai CL, Liang X, Shi Y, Chu PH, Pfaff SL, et al. (2003) Isl1 identifies a cardiac progenitor population that proliferates prior to differentiation and contributes a majority of cells to the heart. Dev Cell 5: 877–889. PubMed PMC

Zaffran S, Kelly RG, Meilhac SM, Buckingham ME, Brown NA (2004) Right ventricular myocardium derives from the anterior heart field. Circ Res 95: 261–268. PubMed

Milani-Nejad N, Janssen PM (2014) Small and large animal models in cardiac contraction research: advantages and disadvantages. Pharmacol Ther 141: 235–249. PubMed PMC

Characterization of a new model of chemotherapy-induced heart failure with reduced ejection fraction and nephrotic syndrome in Ren-2 transgenic rats

MicroRNAs in doxorubicin-induced cardiotoxicity: The DNA damage response

Downregulation of myogenic microRNAs in sub-chronic but not in sub-acute model of daunorubicin-induced cardiomyopathy