Monitoring Mitochondrial Morphology and Respiration in Doxorubicin-Induced Cardiomyopathy

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2022 Jun 30

PMID 35771444

Authors

Chowdhury S Abdullah

Richa Aishwarya

Mahboob Morshed

Naznin Sultana Remex

Sumitra Miriyala

Manikandan Panchatcharam

Md Shenuarin Bhuiyan

Affiliations

Soon will be listed here.

Abstract

Doxorubicin (DOX)-induced cardiomyopathy constitutes dose-dependent cardiac toxicity, culminating in fatal heart failure progression. Cardiac toxicity limits effective and subsequent use of DOX in chemotherapy regimens in pediatric, adult, and recurrent cancer patients. DOX-induced profound alterations in mitochondrial morphology, dynamics, and defects in mitochondrial energy metabolism in the heart comprise key stressors in DOX-induced cardiotoxicity. Hence, the discovery of novel molecular targets and therapeutics to mitigate DOX-induced mitochondrial dysfunctions are imperative. Herein, we provided two laboratory protocols to monitor DOX-induced alterations in mitochondrial morphology and respiration in isolated primary neonatal rat cardiomyocytes. Neonatal rat cardiomyocytes are extensively used to monitor signaling mechanisms regulating cardiomyopathy in vitro. Therefore, these protocols will help researchers study the effects of novel pharmacological and genetic manipulations against DOX-induced alterations in mitochondrial morphology and energy metabolism in cardiomyocytes.

Citing Articles

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References

Davies K, Doroshow J, Hochstein P . Mitochondrial NADH dehydrogenase-catalyzed oxygen radical production by adriamycin, and the relative inactivity of 5-iminodaunorubicin. FEBS Lett. 1983; 153(1):227-30. DOI: 10.1016/0014-5793(83)80153-7. View

Kremer L, van der Pal H, Offringa M, van Dalen E, VOUTE P . Frequency and risk factors of subclinical cardiotoxicity after anthracycline therapy in children: a systematic review. Ann Oncol. 2002; 13(6):819-29. DOI: 10.1093/annonc/mdf167. View

Alam S, Abdullah C, Aishwarya R, Morshed M, Nitu S, Miriyala S . Dysfunctional Mitochondrial Dynamic and Oxidative Phosphorylation Precedes Cardiac Dysfunction in R120G-αB-Crystallin-Induced Desmin-Related Cardiomyopathy. J Am Heart Assoc. 2020; 9(23):e017195. PMC: 7763772. DOI: 10.1161/JAHA.120.017195. View

Lankelma J, Dekker H, Luque F, Luykx S, Hoekman K, van der Valk P . Doxorubicin gradients in human breast cancer. Clin Cancer Res. 1999; 5(7):1703-7. View

Doenst T, Nguyen T, Abel E . Cardiac metabolism in heart failure: implications beyond ATP production. Circ Res. 2013; 113(6):709-24. PMC: 3896379. DOI: 10.1161/CIRCRESAHA.113.300376. View

Sutherland F, Hearse D . The isolated blood and perfusion fluid perfused heart. Pharmacol Res. 2000; 41(6):613-27. DOI: 10.1006/phrs.1999.0653. View

Davies K, Doroshow J . Redox cycling of anthracyclines by cardiac mitochondria. I. Anthracycline radical formation by NADH dehydrogenase. J Biol Chem. 1986; 261(7):3060-7. View

Swain S, Whaley F, Ewer M . Congestive heart failure in patients treated with doxorubicin: a retrospective analysis of three trials. Cancer. 2003; 97(11):2869-79. DOI: 10.1002/cncr.11407. View

Serrano J, Palmeira C, Kuehl D, Wallace K . Cardioselective and cumulative oxidation of mitochondrial DNA following subchronic doxorubicin administration. Biochim Biophys Acta. 1999; 1411(1):201-5. DOI: 10.1016/s0005-2728(99)00011-0. View

10.

Steinherz L, Steinherz P . Delayed cardiac toxicity from anthracycline therapy. Pediatrician. 1991; 18(1):49-52. View

11.

Karukstis K, Thompson E, Whiles J, Rosenfeld R . Deciphering the fluorescence signature of daunomycin and doxorubicin. Biophys Chem. 1998; 73(3):249-63. DOI: 10.1016/s0301-4622(98)00150-1. View

12.

Alam S, Abdullah C, Aishwarya R, Miriyala S, Panchatcharam M, Peretik J . Aberrant Mitochondrial Fission Is Maladaptive in Desmin Mutation-Induced Cardiac Proteotoxicity. J Am Heart Assoc. 2018; 7(14). PMC: 6064863. DOI: 10.1161/JAHA.118.009289. View

13.

Sledge G, Neuberg D, Bernardo P, Ingle J, Martino S, Rowinsky E . Phase III trial of doxorubicin, paclitaxel, and the combination of doxorubicin and paclitaxel as front-line chemotherapy for metastatic breast cancer: an intergroup trial (E1193). J Clin Oncol. 2003; 21(4):588-92. DOI: 10.1200/JCO.2003.08.013. View

14.

Tachibana S, Chen C, Zhang O, Schurr S, Hill C, Li R . Analyzing Oxygen Consumption Rate in Primary Cultured Mouse Neonatal Cardiomyocytes Using an Extracellular Flux Analyzer. J Vis Exp. 2019; (144). PMC: 7605210. DOI: 10.3791/59052. View

15.

Berthiaume J, Wallace K . Persistent alterations to the gene expression profile of the heart subsequent to chronic Doxorubicin treatment. Cardiovasc Toxicol. 2007; 7(3):178-91. DOI: 10.1007/s12012-007-0026-0. View

16.

Vercesi A, Castilho R, Kowaltowski A, de Oliveira H, de Souza-Pinto N, Figueira T . Mitochondrial calcium transport and the redox nature of the calcium-induced membrane permeability transition. Free Radic Biol Med. 2018; 129:1-24. DOI: 10.1016/j.freeradbiomed.2018.08.034. View

17.

Song M, Mihara K, Chen Y, Scorrano L, Dorn 2nd G . Mitochondrial fission and fusion factors reciprocally orchestrate mitophagic culling in mouse hearts and cultured fibroblasts. Cell Metab. 2015; 21(2):273-286. PMC: 4318753. DOI: 10.1016/j.cmet.2014.12.011. View

18.

Duray P, Cuono C, Madri J . Demonstration of cutaneous doxorubicin extravasation by rhodamine-filtered fluorescence microscopy. J Surg Oncol. 1986; 31(1):21-5. DOI: 10.1002/jso.2930310104. View

19.

Dhingra R, Margulets V, Chowdhury S, Thliveris J, Jassal D, Fernyhough P . Bnip3 mediates doxorubicin-induced cardiac myocyte necrosis and mortality through changes in mitochondrial signaling. Proc Natl Acad Sci U S A. 2014; 111(51):E5537-44. PMC: 4280597. DOI: 10.1073/pnas.1414665111. View

20.

Bonadonna G, Monfardini S, De Lena M, Fossati-Bellani F . Clinical evaluation of adriamycin, a new antitumour antibiotic. Br Med J. 1969; 3(5669):503-6. PMC: 1984262. DOI: 10.1136/bmj.3.5669.503. View