title: Doxorubicin impairs cardiomyocyte viability by suppressing transcription factor EB expression and disrupting autophagy.
abstract: Doxorubicin (DOX) is an effective anti-cancer agent. However, DOX treatment increases patient susceptibility to dilated cardiomyopathy. DOX predisposes cardiomyocytes to insult by suppressing mitochondrial energy metabolism, altering calcium flux, and disrupting proteolysis and proteostasis. Prior studies have assessed the role of macroautophagy in DOX cardiotoxicity; however, limited studies have examined whether DOX mediates cardiac injury through dysfunctions in inter- and/or intra-lysosomal signaling events. Lysosomal signaling and function is governed by transcription factor EB (TFEB). In the present study, we hypothesized that DOX caused myocyte injury by impairing lysosomal function and signaling through negative regulation of TFEB. Indeed, we found that DOX repressed cellular TFEB expression, which was associated with impaired cathepsin proteolytic activity across in vivo, ex vivo, and in vitro models of DOX cardiotoxicity. Furthermore, we observed that loss of TFEB was associated with reduction in macroautophagy protein expression, inhibition of autophagic flux, impairments in lysosomal cathepsin B activity, and activation of cell death. Restoration and/or activation of TFEB in DOX-treated cardiomyocytes prevented DOX-induced suppression of cathepsin B activity, reduced DOX-mediated reactive oxygen species (ROS) overproduction, attenuated activation of caspase-3, and improved cellular viability. Collectively, loss of TFEB inhibits lysosomal autophagy, rendering cardiomyocytes susceptible to DOX-induced proteotoxicity and injury. Our data reveal a novel mechanism wherein DOX primes cardiomyocytes for cell death by depleting cellular TFEB.
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS, Canada Dalhousie Medicine New Brunswick, Saint John, NB, Canada.
authors.forename: Jordan J
authors.initials: JJ
authors.lastname: Bartlett
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS, Canada Dalhousie Medicine New Brunswick, Saint John, NB, Canada.
authors.forename: Purvi C
authors.initials: PC
authors.lastname: Trivedi
authors.affiliation: College of Pharmacy and Department of Medicine (Cardiology), Dalhousie University, Halifax, NS, Canada.
authors.forename: Pollen
authors.initials: P
authors.lastname: Yeung
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS, Canada Dalhousie Medicine New Brunswick, Saint John, NB, Canada.
authors.forename: Petra C
authors.initials: PC
authors.lastname: Kienesberger
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS, Canada Dalhousie Medicine New Brunswick, Saint John, NB, Canada.
authors.forename: Thomas
authors.initials: T
authors.lastname: Pulinilkunnil
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001343:Autophagy / Q000187:drug effects*; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004317:Doxorubicin / Q000494:pharmacology*; D015151:Immunoblotting; D020287:In Situ Nick-End Labeling; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D059748:Proteolysis / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D017382:Reactive Oxygen Species; C574124:TFEB protein, rat; D004317:Doxorubicin
keywords: TFEB; autophagy; cardiomyocytes; doxorubicin; heart failure; lysosomes