title: Loss of Mrp1 Potentiates Doxorubicin-Induced Cytotoxicity in Neonatal Mouse Cardiomyocytes and Cardiac Fibroblasts.
abstract: Doxorubicin (DOX) induces dose-dependent cardiotoxicity in part due to its ability to induce oxidative stress. We showed that loss of multidrug resistance-associated protein 1 (Abcc1/Mrp1) potentiates DOX-induced cardiac dysfunction in mice in vivo Here, we characterized DOX toxicity in cultured cardiomyocytes (CM) and cardiac fibroblasts (CF) derived from C57BL wild type (WT) and Mrp1 null (Mrp1-/-) neonatal mice. CM accumulated more intracellular DOX relative to CF but this accumulation did not differ between genotypes. Following DOX (0.3-4 μM), Mrp1-/- CM, and CF, especially CM, showed a greater decrease in viability and increased apoptosis and DNA damage, demonstrated by higher caspase 3 cleavage, poly (ADP-ribose) polymerase 1 (PARP) cleavage and phosphorylated histone H2AX (γH2AX) levels versus WT cells. Saline- and DOX-treated Mrp1-/- cells had significantly higher intracellular GSH and GSSG compared with WT cells (P < .05), but the redox potential (Eh) of the GSH/GSSG pool did not differ between genotypes in CM and CF, indicating that Mrp1-/- cells maintain this major redox couple. DOX increased expression of the rate-limiting GSH synthesis enzyme glutamate-cysteine ligase catalytic (GCLc) and regulatory subunits (GCLm) to a significantly greater extent in Mrp1-/- versus WT cells, suggesting adaptive responses to oxidative stress in Mrp1-/- cells that were inadequate to afford protection. Expression of extracellular superoxide dismutase (ECSOD/SOD3) was lower (P < .05) in Mrp1-/- versus WT CM treated with saline (62% ± 8% of WT) or DOX (43% ± 12% of WT). Thus, Mrp1 protects CM in particular and CF against DOX-induced toxicity, potentially by regulating extracellular redox states.
authors.affiliation: *Department of Toxicology and Cancer Biology, College of Medicine, University of Kentucky, Lexington, Kentucky 40536;
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: *Department of Toxicology and Cancer Biology, College of Medicine, University of Kentucky, Lexington, Kentucky 40536;
authors.forename: Daret
authors.initials: D
authors.lastname: St Clair
authors.affiliation: Department of Chemistry, University of Kentucky, Lexington, Kentucky 40506.
authors.forename: Allan
authors.initials: A
authors.lastname: Butterfield
authors.affiliation: *Department of Toxicology and Cancer Biology, College of Medicine, University of Kentucky, Lexington, Kentucky 40536; maryv@email.uky.edu.
authors.forename: Mary
authors.initials: M
authors.lastname: Vore
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D066126:Cardiotoxicity; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000633:toxicity*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005978:Glutathione / Q000378:metabolism; D006331:Heart Diseases / Q000139:chemically induced* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006657:Histones / Q000378:metabolism; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000172:deficiency* / Q000235:genetics; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013997:Time Factors
chemical_list: D000903:Antibiotics, Antineoplastic; D053148:Caspase 3; D004317:Doxorubicin; D005978:Glutathione; D006657:Histones; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D000071137:Poly (ADP-Ribose) Polymerase-1; D013482:Superoxide Dismutase; C499090:Sod3 protein, mouse; C499321:Parp1 protein, mouse; C505409:Casp3 protein, mouse; C421109:multidrug resistance-associated protein 1; C475961:gamma-H2AX protein, mouse
keywords: ECSOD; Mrp1; doxorubicin; glutathione; glutathione disulfide
references.citation: J Clin Invest. 1996 Sep 1;98(5):1253-60
references.pmid: 8787689
references.citation: Cardiovasc Res. 2005 Jan 1;65(1):40-51
references.pmid: 15621032
references.citation: Free Radic Res. 2008 Aug;42(8):689-706
references.pmid: 18671159
references.citation: Circulation. 2001 Apr 10;103(14):1893-8
references.pmid: 11294809
references.citation: Biochemistry. 2006 Jun 27;45(25):7889-96
references.pmid: 16784241
references.citation: FASEB J. 1998 Apr;12(6):479-86
references.pmid: 9535220
references.citation: PLoS One. 2011;6(11):e28134
references.pmid: 22132230
references.citation: Am J Pathol. 1996 Apr;148(4):1237-47
references.pmid: 8644864
references.citation: J Clin Invest. 2011 Oct;121(10 ):3924-31
references.pmid: 21881209
references.citation: Cardiovasc Res. 2002 Aug 1;55(2):239-49
references.pmid: 12123763
references.citation: Annu Rev Pharmacol Toxicol. 2014;54:95-117
references.pmid: 24050699
references.citation: Basic Res Cardiol. 2005 Jul;100(4):337-45
references.pmid: 15822004
references.citation: J Clin Oncol. 2012 May 1;30(13):1422-8
references.pmid: 21900104
references.citation: Cell. 2007 Aug 10;130(3):427-39
references.pmid: 17693254
references.citation: Am J Physiol Cell Physiol. 2008 Oct;295(4):C849-68
references.pmid: 18684987
references.citation: Circulation. 1998 Oct 6;98(14):1438-48
references.pmid: 9760299
references.citation: J Pharmacol Exp Ther. 2015 Nov;355(2):272-9
references.pmid: 26354996
references.citation: J Biol Chem. 2007 May 11;282(19):14337-47
references.pmid: 17374608
references.citation: J Biol Chem. 1975 Feb 25;250(4):1422-6
references.pmid: 1112810
references.citation: J Am Heart Assoc. 2014 Jun 23;3(3):e000996
references.pmid: 24958779
references.citation: J Mol Cell Cardiol. 2012 Jun;52(6):1213-25
references.pmid: 22465037
references.citation: Histopathology. 2009 May;54(6):657-66
references.pmid: 19438741
references.citation: Neuroscience. 2004;129(2):349-60
references.pmid: 15501592
references.citation: PLoS One. 2012;7(3):e33420
references.pmid: 22442691
references.citation: J Biol Chem. 2007 Oct 19;282(42):30452-65
references.pmid: 17724027
references.citation: Circ Res. 2009 Dec 4;105(12):1164-76
references.pmid: 19959782
references.citation: Am J Physiol Gastrointest Liver Physiol. 2010 Aug;299(2):G523-30
references.pmid: 20466942
references.citation: Pharm Res. 2005 Jun;22(6):842-6
references.pmid: 15948026
references.citation: Biochim Biophys Acta. 2015 Jan;1852(1):53-60
references.pmid: 25446995
references.citation: J Clin Oncol. 2005 Dec 1;23(34):8597-605
references.pmid: 16314622
references.citation: Bioessays. 1998 Nov;20(11):931-40
references.pmid: 9872059
references.citation: Cancer Res. 1997 Dec 1;57(23):5238-42
references.pmid: 9393741
references.citation: Am J Physiol Heart Circ Physiol. 2015 Nov;309(9):H1453-67
references.pmid: 26386112
references.citation: Cell Biol Int. 2012 Jan;36(1):79-86
references.pmid: 21929509
references.citation: Nat Med. 1997 Nov;3(11):1275-9
references.pmid: 9359705
references.citation: J Biol Chem. 1993 Sep 15;268(26):19675-80
references.pmid: 8103521
references.citation: J Biol Chem. 2001 Jul 27;276(30):27846-54
references.pmid: 11375986
references.citation: J Pharmacol Exp Ther. 2015 Nov;355(2):280-7
references.pmid: 26354995
references.citation: Hypertension. 2010 Jun;55(6):1412-7
references.pmid: 20439823
references.citation: J Biol Chem. 2003 Mar 7;278(10):8058-64
references.pmid: 12502708
references.citation: Circulation. 2005 Dec 13;112(24):3754-62
references.pmid: 16330681
references.citation: Physiol Genomics. 2007 Dec 19;32(1):74-81
references.pmid: 17895394
references.citation: J Mol Cell Cardiol. 2009 Nov;47(5):730-42
references.pmid: 19695260
references.citation: J Biol Chem. 2012 Jan 2;287(1):607-18
references.pmid: 22084240
references.citation: J Mol Cell Cardiol. 1992 Jul;24(7):669-81
references.pmid: 1404407
references.citation: Toxicol In Vitro. 2010 Sep;24(6):1584-91
references.pmid: 20600803
references.citation: Br J Haematol. 2005 Feb;128(3):324-32
references.pmid: 15667534
references.citation: Biochem Pharmacol. 2009 Jan 1;77(1):76-85
references.pmid: 18840413
references.citation: Circ Res. 2005 Sep 30;97(7):637-44
references.pmid: 16123331
references.citation: Free Radic Res Commun. 1992;17(1):41-7
references.pmid: 1446846
references.citation: Mol Pharmacol. 1997 Sep;52(3):344-53
references.pmid: 9281595
references.citation: Free Radic Biol Med. 2006 Nov 1;41(9):1470-7
references.pmid: 17023274
references.citation: Antioxid Redox Signal. 2013 Mar 20;18(9):1114-27
references.pmid: 22938199
references.citation: Antioxid Redox Signal. 2011 Mar 15;14 (6):957-71
references.pmid: 21126175
references.citation: Basic Res Cardiol. 2012 Nov;107(6):305
references.pmid: 23099819
references.citation: Biochem Pharmacol. 2007 Jun 1;73(11):1727-37
references.pmid: 17359940
references.citation: J Biol Chem. 2003 Nov 21;278(47):46369-77
references.pmid: 12954617
references.citation: Toxicol Appl Pharmacol. 2004 Feb 15;195(1):12-22
references.pmid: 14962501
references.citation: FASEB J. 2012 Apr;26(4):1442-51
references.pmid: 22202674
references.citation: Apoptosis. 2014 Jan;19(1):117-34
references.pmid: 24146141
references.citation: Mol Aspects Med. 2009 Feb-Apr;30(1-2):86-98
references.pmid: 18812186
references.citation: Arch Immunol Ther Exp (Warsz). 2009 Nov-Dec;57(6):435-45
references.pmid: 19866340
references.citation: J Biol Chem. 2014 Nov 7;289(45):30880-8
references.pmid: 25281745