Doxorubicin cardiomyopathy is a lethal pathology characterized by oxidative stress, mitochondrial dysfunction, and contractile impairment, leading to cell death. Although extensive research has been done to understand the pathophysiology of doxorubicin cardiomyopathy, no effective treatments are available. We investigated whether monoamine oxidases (MAOs) could be involved in doxorubicin-derived oxidative stress, and in the consequent mitochondrial, cardiomyocyte, and cardiac dysfunction.
Results:
We used neonatal rat ventricular myocytes (NRVMs) and adult mouse ventricular myocytes (AMVMs). Doxorubicin alone (i.e., 0.5 μM doxorubicin) or in combination with H2O2 induced an increase in mitochondrial formation of reactive oxygen species (ROS), which was prevented by the pharmacological inhibition of MAOs in both NRVMs and AMVMs. The pharmacological approach was supported by the genetic ablation of MAO-A in NRVMs. In addition, doxorubicin-derived ROS caused lipid peroxidation and alterations in mitochondrial function (i.e., mitochondrial membrane potential, permeability transition, redox potential), mitochondrial morphology (i.e., mitochondrial distribution and perimeter), sarcomere organization, intracellular [Ca2+] homeostasis, and eventually cell death. All these dysfunctions were abolished by MAO inhibition. Of note, in vivo MAO inhibition prevented chamber dilation and cardiac dysfunction in doxorubicin-treated mice.
Innovation and Conclusion:
This study demonstrates that the severe oxidative stress induced by doxorubicin requires the involvement of MAOs, which modulate mitochondrial ROS generation. MAO inhibition provides evidence that mitochondrial ROS formation is causally linked to all disorders caused by doxorubicin in vitro and in vivo. Based upon these results, MAO inhibition represents a novel therapeutic approach for doxorubicin cardiomyopathy.
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References
1.
AliMK, EwerMS, GibbsHR, SwaffordJ, and GraffKL. Late doxorubicin-associated cardiotoxicity in children. The possible role of intercurrent viral infection. Cancer, 74: 182–188, 1994.
2.
AntonucciS, MulveyJF, BurgerN, Di SanteM, HallAR, HinchyEC, CaldwellST, GruszczykAV, DeshwalS, HartleyRC, KaludercicN, MurphyMP, Di LisaF, and KriegT. Selective mitochondrial superoxide generation in vivo is cardioprotective through hormesis. Free Radic Biol Med, 134: 678–687, 2019.
3.
BassaniJW, BassaniRA, and BersDM. Twitch-dependent SR Ca accumulation and release in rabbit ventricular myocytes. Am J Physiol, 265: C533–C540, 1993.
BindaC, Newton-VinsonP, HubalekF, EdmondsonDE, and MatteviA. Structure of human monoamine oxidase B, a drug target for the treatment of neurological disorders. Nat Struct Biol, 9: 22–26, 2002.
8.
Cadeddu DessalviC, PepeA, PennaC, GimelliA, MadonnaR, MeleD, MonteI, NovoG, NugaraC, ZitoC, MoslehiJJ, de BoerRA, LyonAR, TocchettiCG, and MercuroG. Sex differences in anthracycline-induced cardiotoxicity: the benefits of estrogens. Heart Fail Rev, 24: 915–925, 2019.
9.
CappettaD, De AngelisA, SapioL, PreziosoL, IllianoM, QuainiF, RossiF, BerrinoL, NaviglioS, and UrbanekK. Oxidative stress and cellular response to doxorubicin: a common factor in the complex milieu of anthracycline cardiotoxicity. Oxid Med Cell Longev, 2017: 1521020, 2017.
10.
CardinaleD, ColomboA, LamantiaG, ColomboN, CivelliM, De GiacomiG, RubinoM, VegliaF, FiorentiniC, and CipollaCM. Anthracycline-induced cardiomyopathy: clinical relevance and response to pharmacologic therapy. J Am Coll Cardiol, 55: 213–220, 2010.
11.
ChandelNS.Mitochondria as signaling organelles. BMC Biol, 12: 34, 2014.
12.
ChenMH, ColanSD, and DillerL. Cardiovascular disease: cause of morbidity and mortality in adult survivors of childhood cancers. Circ Res, 108: 619–628, 2011.
13.
CiscatoF, FiladiR, MasgrasI, PizziM, MarinO, DamianoN, PizzoP, GoriA, FrezzatoF, ChiaraF, TrentinL, BernardiP, and RasolaA.Hexokinase 2 displacement from mitochondria-associated membranes prompts Ca(2+) -dependent death of cancer cells. EMBO Rep. e49117, 2020. [Epub ahead of print]; DOI: 10.15252/embr.201949117.
14.
CiscatoF, SciacovelliM, VillanoG, TuratoC, BernardiP, RasolaA, and PontissoP. SERPINB3 protects from oxidative damage by chemotherapeutics through inhibition of mitochondrial respiratory complex I. Oncotarget, 5: 2418–2427, 2014.
15.
DengS, KrugerA, KleschyovAL, KalinowskiL, DaiberA, and WojnowskiL. Gp91phox-containing NAD(P)H oxidase increases superoxide formation by doxorubicin and NADPH. Free Radic Biol Med, 42: 466–473, 2007.
16.
DeshwalS, AntonucciS, KaludercicN, and Di LisaF. Measurement of mitochondrial ROS formation. Methods Mol Biol, 1782: 403–418, 2018.
17.
DeshwalS, Di SanteM, Di LisaF, and KaludercicN. Emerging role of monoamine oxidase as a therapeutic target for cardiovascular disease. Curr Opin Pharmacol, 33: 64–69, 2017.
18.
DeshwalS, ForkinkM, HuCH, BuonincontriG, AntonucciS, Di SanteM, MurphyMP, PaolocciN, Mochly-RosenD, KriegT, Di LisaF, and KaludercicN. Monoamine oxidase-dependent endoplasmic reticulum-mitochondria dysfunction and mast cell degranulation lead to adverse cardiac remodeling in diabetes. Cell Death Differ, 25: 1671–1685, 2018.
19.
Di LisaF, MenaboR, CantonM, BarileM, and BernardiP. Opening of the mitochondrial permeability transition pore causes depletion of mitochondrial and cytosolic NAD+ and is a causative event in the death of myocytes in postischemic reperfusion of the heart. J Biol Chem, 276: 2571–2575, 2001.
EschenhagenT, ForceT, EwerMS, de KeulenaerGW, SuterTM, AnkerSD, AvkiranM, de AzambujaE, BalligandJL, BrutsaertDL, CondorelliG, HansenA, HeymansS, HillJA, HirschE, Hilfiker-KleinerD, JanssensS, de JongS, NeubauerG, PieskeB, PonikowskiP, PirmohamedM, RauchhausM, SawyerD, SugdenPH, WojtaJ, ZannadF, and ShahAM. Cardiovascular side effects of cancer therapies: a position statement from the Heart Failure Association of the European Society of Cardiology. Eur J Heart Fail, 13: 1–10, 2011.
22.
FajardoG, ZhaoM, PowersJ, and BernsteinD. Differential cardiotoxic/cardioprotective effects of beta-adrenergic receptor subtypes in myocytes and fibroblasts in doxorubicin cardiomyopathy. J Mol Cell Cardiol, 40: 375–383, 2006.
23.
FerransVJ, ClarkJR, ZhangJ, YuZX, and HermanEH. Pathogenesis and prevention of doxorubicin cardiomyopathy. Tsitologiia, 39: 928–937, 1997.
24.
FortmannSP, BurdaBU, SengerCA, LinJS, and WhitlockEP. Vitamin and mineral supplements in the primary prevention of cardiovascular disease and cancer: an updated systematic evidence review for the U.S. Preventive Services Task Force. Ann Intern Med, 159: 824–834, 2013.
25.
GeisbergCA and SawyerDB. Mechanisms of anthracycline cardiotoxicity and strategies to decrease cardiac damage. Curr Hypertens Rep, 12: 404–410, 2010.
26.
GlytsouC, CalvoE, CogliatiS, MehrotraA, AnastasiaI, RigoniG, RaimondiA, ShintaniN, LoureiroM, VazquezJ, PellegriniL, EnriquezJA, ScorranoL, and SorianoME. Optic atrophy 1 is epistatic to the core MICOS component MIC60 in mitochondrial cristae shape control. Cell Rep, 17: 3024–3034, 2016.
27.
GordonRR, WuM, HuangCY, HarrisWP, SimHG, LucasJM, ColemanI, HiganoCS, GulatiR, TrueLD, VessellaR, LangePH, GarzottoM, BeerTM, and NelsonPS. Chemotherapy-induced monoamine oxidase expression in prostate carcinoma functions as a cytoprotective resistance enzyme and associates with clinical outcomes. PLoS One, 9: e104271, 2014.
28.
GutscherM, PauleauAL, MartyL, BrachT, WabnitzGH, SamstagY, MeyerAJ, and DickTP. Real-time imaging of the intracellular glutathione redox potential. Nat Methods, 5: 553–559, 2008.
29.
HaoJ, LiWW, DuH, ZhaoZF, LiuF, LuJC, YangXC, and CuiW. Role of vitamin C in cardioprotection of ischemia/reperfusion injury by activation of mitochondrial KATP channel. Chem Pharm Bull (Tokyo), 64: 548–557, 2016.
30.
HegstadAC, AntonsenOH, and YtrehusK. Low concentrations of hydrogen peroxide improve post-ischaemic metabolic and functional recovery in isolated perfused rat hearts. J Mol Cell Cardiol, 29: 2779–2787, 1997.
IkedaS, MatsushimaS, OkabeK, IkedaM, IshikitaA, TadokoroT, EnzanN, YamamotoT, SadaM, DeguchiH, MorimotoS, IdeT, and TsutsuiH. Blockade of L-type Ca(2+) channel attenuates doxorubicin-induced cardiomyopathy via suppression of CaMKII-NF-kappaB pathway. Sci Rep, 9: 9850, 2019.
33.
JainD.Cardiotoxicity of doxorubicin and other anthracycline derivatives. J Nucl Cardiol, 7: 53–62, 2000.
34.
KaludercicN, CarpiA, NagayamaT, SivakumaranV, ZhuG, LaiEW, BedjaD, De MarioA, ChenK, GabrielsonKL, LindseyML, PacakK, TakimotoE, ShihJC, KassDA, Di LisaF, and PaolocciN. Monoamine oxidase B prompts mitochondrial and cardiac dysfunction in pressure overloaded hearts. Antioxid Redox Signal, 20: 267–280, 2014.
35.
KaludercicN, Mialet-PerezJ, PaolocciN, PariniA, and Di LisaF. Monoamine oxidases as sources of oxidants in the heart. J Mol Cell Cardiol, 73: 34–42, 2014.
36.
KaludercicN, TakimotoE, NagayamaT, FengN, LaiEW, BedjaD, ChenK, GabrielsonKL, BlakelyRD, ShihJC, PacakK, KassDA, Di LisaF, and PaolocciN. Monoamine oxidase A-mediated enhanced catabolism of norepinephrine contributes to adverse remodeling and pump failure in hearts with pressure overload. Circ Res, 106: 193–202, 2010.
37.
KokoszkaJE, CoskunP, EspositoLA, and WallaceDC. Increased mitochondrial oxidative stress in the Sod2 (+/-) mouse results in the age-related decline of mitochondrial function culminating in increased apoptosis. Proc Natl Acad Sci U S A, 98: 2278–2283, 2001.
LiPC, SiddiqiIN, MottokA, LooEY, WuCH, CozenW, SteidlC, and ShihJC. Monoamine oxidase A is highly expressed in classical Hodgkin lymphoma. J Pathol, 243: 220–229, 2017.
40.
LimCC, ZuppingerC, GuoX, KusterGM, HelmesM, EppenbergerHM, SuterTM, LiaoR, and SawyerDB. Anthracyclines induce calpain-dependent titin proteolysis and necrosis in cardiomyocytes. J Biol Chem, 279: 8290–8299, 2004.
41.
LipshultzSE, ScullyRE, LipsitzSR, SallanSE, SilvermanLB, MillerTL, BarryEV, AsselinBL, AthaleU, ClavellLA, LarsenE, MoghrabiA, SamsonY, MichonB, SchorinMA, CohenHJ, NeubergDS, OravEJ, and ColanSD. Assessment of dexrazoxane as a cardioprotectant in doxorubicin-treated children with high-risk acute lymphoblastic leukaemia: long-term follow-up of a prospective, randomised, multicentre trial. Lancet Oncol, 11: 950–961, 2010.
42.
MeinersB, ShenoyC, and ZordokyBN. Clinical and preclinical evidence of sex-related differences in anthracycline-induced cardiotoxicity. Biol Sex Differ, 9: 38, 2018.
43.
MileiJ, BoverisA, LlesuyS, MolinaHA, StorinoR, OrtegaD, and MileiSE. Amelioration of adriamycin-induced cardiotoxicity in rabbits by prenylamine and vitamins A and E. Am Heart J, 111: 95–102, 1986.
44.
MinottiG, MennaP, SalvatorelliE, CairoG, and GianniL. Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev, 56: 185–229, 2004.
45.
MoulinM, PiquereauJ, MateoP, FortinD, Rucker-MartinC, GressetteM, LefebvreF, GresikovaM, SolgadiA, VekslerV, GarnierA, and Ventura-ClapierR. Sexual dimorphism of doxorubicin-mediated cardiotoxicity: potential role of energy metabolism remodeling. Circ Heart Fail, 8: 98–108, 2015.
46.
Munoz-CastanedaJR, MontillaP, MunozMC, BujalanceI, MuntaneJ, and TunezI. Effect of 17-beta-estradiol administration during adriamycin-induced cardiomyopathy in ovariectomized rat. Eur J Pharmacol, 523: 86–92, 2005.
OctaviaY, TocchettiCG, GabrielsonKL, JanssensS, CrijnsHJ, and MoensAL. Doxorubicin-induced cardiomyopathy: from molecular mechanisms to therapeutic strategies. J Mol Cell Cardiol, 52: 1213–1225, 2012.
49.
PalS, AhirM, and SilPC. Doxorubicin-induced neurotoxicity is attenuated by a 43-kD protein from the leaves of Cajanus indicus L. via NF-kappaB and mitochondria dependent pathways. Free Radic Res, 46: 785–798, 2012.
50.
PetronilliV, MiottoG, CantonM, BriniM, ColonnaR, BernardiP, and Di LisaF. Transient and long-lasting openings of the mitochondrial permeability transition pore can be monitored directly in intact cells by changes in mitochondrial calcein fluorescence. Biophys J, 76: 725–734, 1999.
51.
RiccioG, AntonucciS, CoppolaC, D'AvinoC, PiscopoG, FioreD, MaureaC, RussoM, ReaD, ArraC, CondorelliG, Di LisaF, TocchettiCG, De LorenzoC, and MaureaN. Ranolazine attenuates trastuzumab-induced heart dysfunction by modulating ROS production. Front Physiol, 9: 38, 2018.
52.
RistowM.Unraveling the truth about antioxidants: mitohormesis explains ROS-induced health benefits. Nat Med, 20: 709–711, 2014.
53.
SagCM, KohlerAC, AndersonME, BacksJ, and MaierLS. CaMKII-dependent SR Ca leak contributes to doxorubicin-induced impaired Ca handling in isolated cardiac myocytes. J Mol Cell Cardiol, 51: 749–759, 2011.
54.
SalvatorelliE, GuarnieriS, MennaP, LiberiG, CalafioreAM, MariggioMA, MordenteA, GianniL, and MinottiG. Defective one- or two-electron reduction of the anticancer anthracycline epirubicin in human heart. Relative importance of vesicular sequestration and impaired efficiency of electron addition. J Biol Chem, 281: 10990–11001, 2006.
55.
SchindelinJ, Arganda-CarrerasI, FriseE, KaynigV, LongairM, PietzschT, PreibischS, RuedenC, SaalfeldS, SchmidB, TinevezJY, WhiteDJ, HartensteinV, EliceiriK, TomancakP, and CardonaA. Fiji: an open-source platform for biological-image analysis. Nat Methods, 9: 676–682, 2012.
56.
SenaLA and ChandelNS. Physiological roles of mitochondrial reactive oxygen species. Mol Cell, 48: 158–167, 2012.
57.
SonSY, MaJ, KondouY, YoshimuraM, YamashitaE, and TsukiharaT. Structure of human monoamine oxidase A at 2.2-A resolution: the control of opening the entry for substrates/inhibitors. Proc Natl Acad Sci U S A, 105: 5739–5744, 2008.
58.
SoratoE, MenazzaS, ZulianA, SabatelliP, GualandiF, MerliniL, BonaldoP, CantonM, BernardiP, and Di LisaF. Monoamine oxidase inhibition prevents mitochondrial dysfunction and apoptosis in myoblasts from patients with collagen VI myopathies. Free Radic Biol Med, 75: 40–47, 2014.
59.
SpinazziM, CazzolaS, BortolozziM, BaraccaA, LoroE, CasarinA, SolainiG, SgarbiG, CasalenaG, CenacchiG, MalenaA, FrezzaC, CarraraF, AngeliniC, ScorranoL, SalviatiL, and VerganiL. A novel deletion in the GTPase domain of OPA1 causes defects in mitochondrial morphology and distribution, but not in function. Hum Mol Genet, 17: 3291–3302, 2008.
60.
SteinbergJS, CohenAJ, WassermanAG, CohenP, and RossAM. Acute arrhythmogenicity of doxorubicin administration. Cancer, 60: 1213–1218, 1987.
61.
SteinbergSF.Oxidative stress and sarcomeric proteins. Circ Res, 112: 393–405, 2013.
62.
SterbaM, PopelovaO, VavrovaA, JirkovskyE, KovarikovaP, GerslV, and SimunekT. Oxidative stress, redox signaling, and metal chelation in anthracycline cardiotoxicity and pharmacological cardioprotection. Antioxid Redox Signal, 18: 899–929, 2013.
63.
SunA, ChengY, ZhangY, ZhangQ, WangS, TianS, ZouY, HuK, RenJ, and GeJ. Aldehyde dehydrogenase 2 ameliorates doxorubicin-induced myocardial dysfunction through detoxification of 4-HNE and suppression of autophagy. J Mol Cell Cardiol, 71: 92–104, 2014.
64.
TebbiCK, LondonWB, FriedmanD, VillalunaD, De AlarconPA, ConstineLS, MendenhallNP, SpostoR, ChauvenetA, and SchwartzCL. Dexrazoxane-associated risk for acute myeloid leukemia/myelodysplastic syndrome and other secondary malignancies in pediatric Hodgkin's disease. J Clin Oncol, 25: 493–500, 2007.
65.
ThornCF, OshiroC, MarshS, Hernandez-BoussardT, McLeodH, KleinTE, and AltmanRB. Doxorubicin pathways: pharmacodynamics and adverse effects. Pharmacogenet Genomics, 21: 440–446, 2011.
66.
TimolatiF, OttD, PentassugliaL, GiraudMN, PerriardJC, SuterTM, and ZuppingerC. Neuregulin-1 beta attenuates doxorubicin-induced alterations of excitation-contraction coupling and reduces oxidative stress in adult rat cardiomyocytes. J Mol Cell Cardiol, 41: 845–854, 2006.
67.
TonoloF, FoldaA, CesaroL, ScalconV, MarinO, FerroS, BindoliA, and RigobelloMP. Milk-derived bioactive peptides exhibit antioxidant activity through the Keap1-Nrf2 signaling pathway. J Funct Foods, 64: 2020.
68.
TonoloF, SandreM, FerroS, FoldaA, ScalconV, ScutariG, FellerE, MarinO, BindoliA, and RigobelloMP. Milk-derived bioactive peptides protect against oxidative stress in a Caco-2 cell model. Food Funct, 9: 1245–1253, 2018.
69.
WallaceKB, SardaoVA, and OliveiraPJ. Mitochondrial determinants of doxorubicin-induced cardiomyopathy. Circ Res, 126: 926–941, 2020.
70.
WangF, HeQ, SunY, DaiX, and YangXP. Female adult mouse cardiomyocytes are protected against oxidative stress. Hypertension, 55: 1172–1178, 2010.
71.
WuJB, ShaoC, LiX, LiQ, HuP, ShiC, LiY, ChenYT, YinF, LiaoCP, StilesBL, ZhauHE, ShihJC, and ChungLW. Monoamine oxidase A mediates prostate tumorigenesis and cancer metastasis. J Clin Invest, 124: 2891–2908, 2014.
72.
YagiK.Assay for blood plasma or serum. Methods Enzymol, 105: 328–331, 1984.
73.
ZhouS, StarkovA, FrobergMK, LeinoRL, and WallaceKB. Cumulative and irreversible cardiac mitochondrial dysfunction induced by doxorubicin. Cancer Res, 61: 771–777, 2001.
74.
ZouY, YaoA, ZhuW, KudohS, HiroiY, ShimoyamaM, UozumiH, KohmotoO, TakahashiT, ShibasakiF, NagaiR, YazakiY, and KomuroI. Isoproterenol activates extracellular signal-regulated protein kinases in cardiomyocytes through calcineurin. Circulation, 104: 102–108, 2001.
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