Elamipretide protects H9c2 Rat Cardiomyoblasts against Doxorubicin-Induced Disruption of Mitochondrial Quality Control by Restoration of Fusion and Fission Balance

Authors

  • Marwa Mahmoud College of Pharmacy, Mustansiriyah University
  • Inaam S. Arif Department of Pharmacology and Toxicology, Collage of pharmacy, Mustansiriyah University, Iraq, Baghdad
  • Muthanna I. AL-Ezzi Department of Pharmacology and Toxicology, College of Pharmacy, Mustansiriyah University, Iraq

DOI:

https://doi.org/10.31351/vol33iss2pp49-57

Keywords:

doxorubicin, elamipretide, MFN2, DRP1, H9c2 cell line

Abstract

Doxorubicin (DOX) has been used to treat malignant diseases for over 40 years. The main constraint to its clinical application is dose-dependent cardiotoxicity which is regarded as a major matter of concern that limits its medical usefulness. Mitochondrial dysfunction is considered the chief contributor to DOX-induced cardiotoxicity and it involves disruption of mitochondrial quality control, mainly impaired fusion, and enhanced fission processes. Compounds that specifically target the mitochondria and restore fusion and fission balance are considered a promising tool to protect or treat cardiomyopathy and heart failure and thus could be investigated as a novel strategy to alleviate DOX-induced cardiac toxicity, one of which is elamipretide (ELAM). In this study, MTT assay revealed that DOX induces a significant reduction in H9c2 cell viability which is both time and dose-dependent whereas ELAM has no significant effect on the viability of the relevant cells at most of the concentrations used. Additionally, western blot analysis showed a significant reduction in the expression of fusion protein MFN2 in the DOX-treated group compared to the control (p***< 0.001) whereas the fission protein DRP1 was significantly upregulated in DOX-treated cells compared to the control (p**< 0.01) and normalization of both proteins was achieved when 10 µM ELAM introduced 48-hour prior to DOX therapy. In conclusion, ELAM could exert an interesting cardioprotective role against DOX-induced cardiotoxicity by restoration of mitochondrial fusion and fission balance.

 

Author Biographies

  • Inaam S. Arif, Department of Pharmacology and Toxicology, Collage of pharmacy, Mustansiriyah University, Iraq, Baghdad

     

     

  • Muthanna I. AL-Ezzi, Department of Pharmacology and Toxicology, College of Pharmacy, Mustansiriyah University, Iraq

     

     

References

Varela-López A, Battino M, Navarro-Hortal MD, Giampieri F, Forbes-Hernández TY, Romero-Márquez JM, et al. An update on the mechanisms related to cell death and toxicity of doxorubicin and the protective role of nutrients. Food Chem Toxicol. 2019 Dec;134:110834.

Cheng D, Tu W, Chen L, Wang H, Wang Q, Liu H, et al. MSCs enhances the protective effects of valsartan on attenuating the doxorubicin-induced myocardial injury via AngII/NOX/ROS/MAPK signaling pathway. Aging. 2021 Sep 29;13(18):22556–70.

He L, Liu F, Li J. Mitochondrial Sirtuins and Doxorubicin-induced Cardiotoxicity. Cardiovasc Toxicol. 2021 Mar;21(3):179–91.

Huang KM, Thomas MZ, Magdy T, Eisenmann ED, Uddin ME, DiGiacomo DF, et al. Targeting OCT3 attenuates doxorubicin-induced cardiac injury. Proc Natl Acad Sci [Internet]. 2021 Feb 2 [cited 2022 Feb 24];118(5). Available from: https://www.pnas.org/content/118/5/e2020168118

Kadowaki H, Akazawa H, Ishida J, Komuro I. Cancer Therapeutics-Related Cardiac Dysfunction ― Insights From Bench and Bedside of Onco-Cardiology ―. Circ J. 2020 Aug 25;84(9):1446–53.

Chan DC. Mitochondrial Dynamics and Its Involvement in Disease. Annu Rev Pathol Mech Dis. 2020 Jan 24;15(1):235–59.

Bisaccia G. Mitochondrial Dysfunction and Heart Disease: Critical Appraisal of an Overlooked Association. 2021. 2021;22(614):19.

Haileselassie B, Mukherjee R, Joshi AU, Napier BA, Massis LM, Ostberg NP, et al. Drp1/Fis1 interaction mediates mitochondrial dysfunction in septic cardiomyopathy. J Mol Cell Cardiol. 2019 May;130:160–9.

Wang J, Toan S, Zhou H. Mitochondrial quality control in cardiac microvascular ischemia-reperfusion injury: New insights into the mechanisms and therapeutic potentials. Pharmacol Res. 2020 Jun;156:104771.

Whitley BN, Engelhart EA, Hoppins S. Mitochondrial dynamics and their potential as a therapeutic target. Mitochondrion. 2019 Nov;49:269–83.

Tilokani L, Nagashima S, Paupe V, Prudent J. Mitochondrial dynamics: overview of molecular mechanisms. Garone C, Minczuk M, editors. Essays Biochem. 2018 Jul 20;62(3):341–60.

Hernandez‐Resendiz S, Prunier F, Girao H, Dorn G, Hausenloy DJ, EU‐CARDIOPROTECTION COST Action (CA16225). Targeting mitochondrial fusion and fission proteins for cardioprotection. J Cell Mol Med. 2020 Jun;24(12):6571–85.

Givvimani S, Pushpakumar S, Veeranki S, Tyagi SC. Dysregulation of Mfn2 and Drp-1 proteins in heart failure. Can J Physiol Pharmacol. 2014 Jul;92(7):583–91.

Jin J yu, Wei X xiang, Zhi X ling, Wang X hong, Meng D. Drp1-dependent mitochondrial fission in cardiovascular disease. Acta Pharmacol Sin. 2021 May;42(5):655–64.

Zemirli N, Morel E, Molino D. Mitochondrial Dynamics in Basal and Stressful Conditions. Int J Mol Sci. 2018 Feb 13;19(2):564.

Osataphan N. Effects of doxorubicin-induced cardiotoxicity on cardiac mitochondrial dynamics and mitochondrial function: Insights for future interventions. 2019. 2019;24.

Yin Y, Shen H. Advances in Cardiotoxicity Induced by Altered Mitochondrial Dynamics and Mitophagy. Front Cardiovasc Med [Internet]. 2021 [cited 2022 Mar 9];8. Available from: https://www.frontiersin.org/article/10.3389/fcvm.2021.739095

Du J, Hang P, Pan Y, Feng B, Zheng Y, Chen T, et al. Inhibition of miR-23a attenuates doxorubicin-induced mitochondria-dependent cardiomyocyte apoptosis by targeting the PGC-1α/Drp1 pathway. Toxicol Appl Pharmacol. 2019 Apr;369:73–81.

Huang J, Wu R, Chen L, Yang Z, Yan D, Li M. Understanding Anthracycline Cardiotoxicity From Mitochondrial Aspect. Front Pharmacol [Internet]. 2022 [cited 2022 Mar 2];13. Available from: https://www.frontiersin.org/article/10.3389/fphar.2022.811406

Szeto HH. First-in-class cardiolipin-protective compound as a therapeutic agent to restore mitochondrial bioenergetics. Br J Pharmacol. 2014;171(8):2029–50.

Peoples JN, Saraf A, Ghazal N, Pham TT, Kwong JQ. Mitochondrial dysfunction and oxidative stress in heart disease. Exp Mol Med. 2019 Dec;51(12):1–13.

Chiao YA, Zhang H, Sweetwyne M, Whitson J, Ting YS, Basisty N, et al. Late-life restoration of mitochondrial function reverses cardiac dysfunction in old mice. eLife. 2020 Jul 10;9:e55513.

Wasmus C, Dudek J. Metabolic Alterations Caused by Defective Cardiolipin Remodeling in Inherited Cardiomyopathies. Life. 2020 Nov 11;10(11):277.

Sabbah HN. Targeting the Mitochondria in Heart Failure: A Translational Perspective. JACC Basic Transl Sci. 2020 Jan 1;5(1):88–106.

Mitchell W, Ng EA, Tamucci JD, Boyd K, Sathappa M, Coscia A, et al. Molecular Mechanism of Action of Mitochondrial Therapeutic SS-31 (Elamipretide): Membrane Interactions and Effects on Surface Electrostatics [Internet]. bioRxiv; 2019 [cited 2022 Mar 14]. p. 735001. Available from: https://www.biorxiv.org/content/10.1101/735001v1

Nickel A, Kohlhaas M, Maack C. Mitochondrial reactive oxygen species production and elimination. J Mol Cell Cardiol. 2014 Aug;73:26–33.

sabbah hani. Chronic Therapy With Elamipretide (MTP-131), a Novel Mitochondria-Targeting Peptide, Improves Left Ventricular and Mitochondrial Function in Dogs With Advanced Heart Failure. 2016.

Li L, Li J, Wang Q, Zhao X, Yang D, Niu L, et al. Shenmai Injection Protects Against Doxorubicin-Induced Cardiotoxicity via Maintaining Mitochondrial Homeostasis. Front Pharmacol [Internet]. 2020 [cited 2023 Jan 6];11. Available from: https://www.frontiersin.org/articles/10.3389/fphar.2020.00815

Sacks B, Onal H, Martorana R, Sehgal A, Harvey A, Wastella C, et al. Mitochondrial targeted antioxidants, mitoquinone and SKQ1, not vitamin C, mitigate doxorubicin-induced damage in H9c2 myoblast: pretreatment vs. co-treatment. BMC Pharmacol Toxicol. 2021 Dec;22(1):49.

Sun MY, Ma DS, Zhao S, Wang L, Ma CY, Bai Y. Salidroside mitigates hypoxia/reoxygenation injury by alleviating endoplasmic reticulum stress‑induced apoptosis in H9c2 cardiomyocytes. Mol Med Rep. 2018 Oct 1;18(4):3760–8.

Zhang Z, Qin X, Wang Z, Li Y, Chen F, Chen R, et al. Oxymatrine pretreatment protects H9c2 cardiomyocytes from hypoxia/reoxygenation injury by modulating the PI3K/Akt pathway. Exp Ther Med. 2021 Jun 1;21(6):1–12.

Zhou F, Li S, Yang J, Ding J, He C, Teng L. In-vitro cardiovascular protective activity of a new achillinoside from Achillea alpina. Rev Bras Farmacogn. 2019 Jul;29(4):445–8.

Kowalczyk A, Czerniawska Piątkowska E. Antioxidant effect of Elamipretide on bull’s sperm cells during freezing/thawing process. Andrology. 2021 Jul;9(4):1275–81.

Checa J, Aran JM. Reactive Oxygen Species: Drivers of Physiological and Pathological Processes. J Inflamm Res. 2020 Dec 2;13:1057–73.

Catanzaro MP, Weiner A, Kaminaris A, Li C, Cai F, Zhao F, et al. Doxorubicin-induced cardiomyocyte death is mediated by unchecked mitochondrial fission and mitophagy. FASEB J. 2019;33(10):11096–108.

Ding M, Shi R, Cheng S, Li M, De D, Liu C, et al. Mfn2-mediated mitochondrial fusion alleviates doxorubicin-induced cardiotoxicity with enhancing its anticancer activity through metabolic switch. Redox Biol. 2022 Jun 1;52:102311.

Tang H, Tao A, Song J, Liu Q, Wang H, Rui T. Doxorubicin-induced cardiomyocyte apoptosis: Role of mitofusin 2. Int J Biochem Cell Biol. 2017 Jul 1;88:55–9.

Emery JM, Ortiz RM. Mitofusin 2: A link between mitochondrial function and substrate metabolism? Mitochondrion. 2021 Nov 1;61:125–37.

Carrasco R, Castillo RL, Gormaz JG, Carrillo M, Thavendiranathan P. Role of Oxidative Stress in the Mechanisms of Anthracycline-Induced Cardiotoxicity: Effects of Preventive Strategies. Oxid Med Cell Longev. 2021 Jan 27;2021:e8863789.

Najafi M, Hooshangi Shayesteh MR, Mortezaee K, Farhood B, Haghi-Aminjan H. The role of melatonin on doxorubicin-induced cardiotoxicity: A systematic review. Life Sci. 2020 Jan 15;241:117173.

Bu X, Wu D, Lu X, Yang L, Xu X, Wang J, et al. Role of SIRT1/PGC-1α in mitochondrial oxidative stress in autistic spectrum disorder. Neuropsychiatr Dis Treat. 2017 Jun 23;13:1633–45.

Wang(a) J, Zhang J, Xiao M, Wang S, Wang(b) J, Guo Y, et al. Molecular mechanisms of doxorubicin-induced cardiotoxicity: novel roles of sirtuin 1-mediated signaling pathways. Cell Mol Life Sci. 2021 Apr 1;78(7):3105–25.

Clayton ZS, Hutton DA, Mahoney SA, Seals DR. Anthracycline chemotherapy-mediated vascular dysfunction as a model of accelerated vascular aging. Aging Cancer. 2021;2(1–2):45–69.

Khayatan D, Razavi SM, Arab ZN, Khanahmadi M, Momtaz S, Butler AE, et al. Regulatory Effects of Statins on SIRT1 and Other Sirtuins in Cardiovascular Diseases. Life. 2022 May;12(5):760.

Zhao H, Li H, Hao S, Chen J, Wu J, Song C, et al. Peptide SS-31 upregulates frataxin expression and improves the quality of mitochondria: implications in the treatment of Friedreich ataxia. Sci Rep. 2017 Aug 29;7(1):9840.

Yang Y, Yang J, Yu Q, Gao Y, Zheng Y, Han L, et al. Regulation of yak longissimus lumborum energy metabolism and tenderness by the AMPK/SIRT1 signaling pathways during postmortem storage. PLOS ONE. 2022 Nov 28;17(11):e0277410.

Song S, Chu L, Liang H, Chen J, Liang J, Huang Z, et al. Protective Effects of Dioscin Against Doxorubicin-Induced Hepatotoxicity Via Regulation of Sirt1/FOXO1/NF-κb Signal. Front Pharmacol [Internet]. 2019 [cited 2023 Jan 20];10. Available from: https://www.frontiersin.org/articles/10.3389/fphar.2019.01030

sabbah. Abnormalities of Mitochondrial Dynamics in the Failing Heart: Normalization Following Long-Term Therapy with Elamipretide. 2018.

Zheng M, Bai Y, Sun X, Fu R, Liu L, Liu M, et al. Resveratrol Reestablishes Mitochondrial Quality Control in Myocardial Ischemia/Reperfusion Injury through Sirt1/Sirt3-Mfn2-Parkin-PGC-1α Pathway. Molecules. 2022 Jan;27(17):5545.

Kai J, Yang X, Wang Z, Wang F, Jia Y, Wang S, et al. Oroxylin a promotes PGC-1α/Mfn2 signaling to attenuate hepatocyte pyroptosis via blocking mitochondrial ROS in alcoholic liver disease. Free Radic Biol Med. 2020 Jun 1;153:89–102.

Rius-Pérez S, Torres-Cuevas I, Millán I, Ortega ÁL, Pérez S. PGC-1α, Inflammation, and Oxidative Stress: An Integrative View in Metabolism. Oxid Med Cell Longev. 2020 Mar 9;2020:e1452696.

Wang J, Lin X, Zhao N, Dong G, Wu W, Huang K, et al. Effects of Mitochondrial Dynamics in the Pathophysiology of Obesity. Front Biosci-Landmark. 2022 Mar 18;27(3):107.

Schirone L, D’Ambrosio L, Forte M, Genovese R, Schiavon S, Spinosa G, et al. Mitochondria and Doxorubicin-Induced Cardiomyopathy: A Complex Interplay. Cells. 2022 Jan;11(13):2000.

Liang X, Wang S, Wang L, Ceylan AF, Ren J, Zhang Y. Mitophagy inhibitor liensinine suppresses doxorubicin-induced cardiotoxicity through inhibition of Drp1-mediated maladaptive mitochondrial fission. Pharmacol Res. 2020 Jul 1;157:104846.

Lu YT, Li LZ, Yang YL, Yin X, Liu Q, Zhang L, et al. Succinate induces aberrant mitochondrial fission in cardiomyocytes through GPR91 signaling. Cell Death Dis. 2018 Jun 4;9(6):1–14.

Huang CY, Chen JY, Kuo CH, Pai PY, Ho TJ, Chen TS, et al. Mitochondrial ROS-induced ERK1/2 activation and HSF2-mediated AT1R upregulation are required for doxorubicin-induced cardiotoxicity. J Cell Physiol. 2018;233(1):463–75.

Sirangelo I, Sapio L, Ragone A, Naviglio S, Iannuzzi C, Barone D, et al. Vanillin Prevents Doxorubicin-Induced Apoptosis and Oxidative Stress in Rat H9c2 Cardiomyocytes. Nutrients. 2020 Aug;12(8):2317.

Li W, He W, Xia P, Sun W, Shi M, Zhou Y, et al. Total Extracts of Abelmoschus manihot L. Attenuates Adriamycin-Induced Renal Tubule Injury via Suppression of ROS-ERK1/2-Mediated NLRP3 Inflammasome Activation. Front Pharmacol [Internet]. 2019 [cited 2023 Jan 16];10. Available from: https://www.frontiersin.org/articles/10.3389/fphar.2019.00567

Wang Y, Han Z, Xu Z, Zhang J. Protective Effect of Optic Atrophy 1 on Cardiomyocyte Oxidative Stress: Roles of Mitophagy, Mitochondrial Fission, and MAPK/ERK Signaling. Oxid Med Cell Longev. 2021 Jun 8;2021:e3726885.

Whitson JA, Martín-Pérez M, Zhang T, Gaffrey MJ, Merrihew GE, Huang E, et al. Elamipretide (SS-31) treatment attenuates age-associated post-translational modifications of heart proteins. GeroScience [Internet]. 2021 Sep 4 [cited 2021 Sep 9]; Available from: https://doi.org/10.1007/s11357-021-00447-6

Montalvo RN, Doerr V, Min K, Szeto HH, Smuder AJ. Doxorubicin-induced oxidative stress differentially regulates proteolytic signaling in cardiac and skeletal muscle. Am J Physiol-Regul Integr Comp Physiol. 2020 Feb 1;318(2):R227–33.

Kim SR, Erin A, Zhang X, Lerman A, Lerman LO. Mitochondrial protection partly mitigates kidney cellular senescence in swine atherosclerotic renal artery stenosis. Cell Physiol Biochem Int J Exp Cell Physiol Biochem Pharmacol. 2019;52(3):617–32.

Sabbah HN. Barth syndrome cardiomyopathy: targeting the mitochondria with elamipretide. Heart Fail Rev. 2021 Mar 1;26(2):237–53.

Kashatus JA, Nascimento A, Myers LJ, Sher A, Byrne FL, Hoehn KL, et al. Erk2 Phosphorylation of Drp1 Promotes Mitochondrial Fission and MAPK-Driven Tumor Growth. Mol Cell. 2015 Feb 5;57(3):537–51.

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Published

2024-06-28