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

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...

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Autores principales: Kretzschmar, Tom, Bekhite, Mohamed M., Wu, Jasmine M. F., Haase, Daniela, Förster, Martin, Müller, Tina, Nietzsche, Sandor, Westermann, Martin, Franz, Marcus, Gräler, Markus H., Schulze, P. Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8584314/
https://www.ncbi.nlm.nih.gov/pubmed/34769283
http://dx.doi.org/10.3390/ijms222111852
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author Kretzschmar, Tom
Bekhite, Mohamed M.
Wu, Jasmine M. F.
Haase, Daniela
Förster, Martin
Müller, Tina
Nietzsche, Sandor
Westermann, Martin
Franz, Marcus
Gräler, Markus H.
Schulze, P. Christian
author_facet Kretzschmar, Tom
Bekhite, Mohamed M.
Wu, Jasmine M. F.
Haase, Daniela
Förster, Martin
Müller, Tina
Nietzsche, Sandor
Westermann, Martin
Franz, Marcus
Gräler, Markus H.
Schulze, P. Christian
author_sort Kretzschmar, Tom
collection PubMed
description 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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spelling pubmed-85843142021-11-12 Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis Kretzschmar, Tom Bekhite, Mohamed M. Wu, Jasmine M. F. Haase, Daniela Förster, Martin Müller, Tina Nietzsche, Sandor Westermann, Martin Franz, Marcus Gräler, Markus H. Schulze, P. Christian Int J Mol Sci Article 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. MDPI 2021-11-01 /pmc/articles/PMC8584314/ /pubmed/34769283 http://dx.doi.org/10.3390/ijms222111852 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kretzschmar, Tom
Bekhite, Mohamed M.
Wu, Jasmine M. F.
Haase, Daniela
Förster, Martin
Müller, Tina
Nietzsche, Sandor
Westermann, Martin
Franz, Marcus
Gräler, Markus H.
Schulze, P. Christian
Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis
title Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis
title_full Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis
title_fullStr Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis
title_full_unstemmed Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis
title_short Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis
title_sort long-chain and very long-chain ceramides mediate doxorubicin-induced toxicity and fibrosis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8584314/
https://www.ncbi.nlm.nih.gov/pubmed/34769283
http://dx.doi.org/10.3390/ijms222111852
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