Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis

GND
1229199403
ORCID
0000-0002-5296-3955
Affiliation
Department of Internal Medicine I, Division of Cardiology, University Hospital Jena
Kretzschmar, Tom;
GND
1139058487
ORCID
0000-0003-0346-9181
Affiliation
Department of Internal Medicine I, Division of Cardiology, University Hospital Jena
Bekhite, Mohamed M.;
GND
1229198784
Affiliation
Department of Internal Medicine I, Division of Cardiology, University Hospital Jena
Wu, Jasmine M. F.;
GND
130141534
Affiliation
Department of Internal Medicine I, Division of Cardiology, University Hospital Jena
Haase, Daniela;
GND
130040762
ORCID
0000-0002-0744-9834
Affiliation
Department of Internal Medicine I, Division of Cardiology, University Hospital Jena, 07747 Jena, Germany;(T.K.);(M.M.B.);(J.M.F.W.);(D.H.);(M.F.);(M.F.)
Förster, Martin;
GND
1217345760
Affiliation
Department of Anesthesiology and Intensive Care Medicine, University Hospital Jena
Müller, Tina;
GND
12341461X
Affiliation
Center of Electron Microscopy, University Hospital Jena
Nietzsche, Sandor;
GND
1230908374
Affiliation
Center of Electron Microscopy, University Hospital Jena
Westermann, Martin;
GND
133832325 008 piz
ORCID
0000-0001-6543-4684
Affiliation
Department of Internal Medicine I, Division of Cardiology, University Hospital Jena
Franz, Marcus;
GND
122793684
ORCID
0000-0001-6650-7849
Affiliation
Department of Anesthesiology and Intensive Care Medicine, University Hospital Jena, 07745 Jena, Germany;(T.M.);(M.H.G.)
Gräler, Markus H.;
GND
121635244
ORCID
0000-0001-9442-7141
Affiliation
Department of Internal Medicine I, Division of Cardiology, University Hospital Jena
Schulze, P. Christian

Doxorubicin (Dox) is a chemotherapeutic agent with cardiotoxicity associated with profibrotic effects. Dox increases ceramide levels with pro-inflammatory effects, cell death, and fibrosis. The purpose of our study was to identify the underlying ceramide signaling pathways. We aimed to characterize the downstream effects on cell survival, metabolism, and fibrosis. Human fibroblasts (hFSF) were treated with 0.7 µM of Dox or transgenically overexpressed ceramide synthase 2 (FLAG-CerS2). Furthermore, cells were pre-treated with MitoTempo (MT) (2 h, 20 µM) or Fumonisin B1 (FuB) (4 h, 100 µM). Protein expression was measured by Western blot or immunofluorescence (IF). Ceramide levels were determined with mass spectroscopy (MS). Visualizations were conducted using laser scanning microscopy (LSM) or electron microscopy. Mitochondrial activity was measured using seahorse analysis. Dox and CerS2 overexpression increased CerS2 protein expression. Coherently, ceramides were elevated with the highest peak for C24:0. Ceramide- induced mitochondrial ROS production was reduced with MT or FuB preincubation. Mitochondrial homeostasis was reduced and accompanied by reduced ATP production. Our data show that the increase in pro-inflammatory ceramides is an essential contributor to Dox side-effects. The accumulation of ceramides resulted in a lipotoxic shift and subsequently mitochondrial structural and functional damage, which was partially reversible following inhibition of ceramide synthesis.

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