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Using cultured canine cardiac slices to model the autophagic flux with doxorubicin
Chemotherapy-induced impairment of autophagy is implicated in cardiac toxicity induced by anti-cancer drugs. Imperfect translation from rodent models and lack of in vitro models of toxicity has limited investigation of autophagic flux dysregulation, preventing design of novel cardioprotective strate...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10019679/ https://www.ncbi.nlm.nih.gov/pubmed/36928870 http://dx.doi.org/10.1371/journal.pone.0282859 |
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author | Boukhalfa, Asma Robinson, Sally R. Meola, Dawn M. Robinson, Nicholas A. Ling, Lauren A. LaMastro, Joey N. Upshaw, Jenica N. Pulakat, Lakshmi Jaffe, Iris Z. London, Cheryl A. Chen, Howard H. Yang, Vicky K. |
author_facet | Boukhalfa, Asma Robinson, Sally R. Meola, Dawn M. Robinson, Nicholas A. Ling, Lauren A. LaMastro, Joey N. Upshaw, Jenica N. Pulakat, Lakshmi Jaffe, Iris Z. London, Cheryl A. Chen, Howard H. Yang, Vicky K. |
author_sort | Boukhalfa, Asma |
collection | PubMed |
description | Chemotherapy-induced impairment of autophagy is implicated in cardiac toxicity induced by anti-cancer drugs. Imperfect translation from rodent models and lack of in vitro models of toxicity has limited investigation of autophagic flux dysregulation, preventing design of novel cardioprotective strategies based on autophagy control. Development of an adult heart tissue culture technique from a translational model will improve investigation of cardiac toxicity. We aimed to optimize a canine cardiac slice culture system for exploration of cancer therapy impact on intact cardiac tissue, creating a translatable model that maintains autophagy in culture and is amenable to autophagy modulation. Canine cardiac tissue slices (350 μm) were generated from left ventricular free wall collected from euthanized client-owned dogs (n = 7) free of cardiovascular disease at the Foster Hospital for Small Animals at Tufts University. Cell viability and apoptosis were quantified with MTT assay and TUNEL staining. Cardiac slices were challenged with doxorubicin and an autophagy activator (rapamycin) or inhibitor (chloroquine). Autophagic flux components (LC3, p62) were quantified by western blot. Cardiac slices retained high cell viability for >7 days in culture and basal levels of autophagic markers remained unchanged. Doxorubicin treatment resulted in perturbation of the autophagic flux and cell death, while rapamycin co-treatment restored normal autophagic flux and maintained cell survival. We developed an adult canine cardiac slice culture system appropriate for studying the effects of autophagic flux that may be applicable to drug toxicity evaluations. |
format | Online Article Text |
id | pubmed-10019679 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-100196792023-03-17 Using cultured canine cardiac slices to model the autophagic flux with doxorubicin Boukhalfa, Asma Robinson, Sally R. Meola, Dawn M. Robinson, Nicholas A. Ling, Lauren A. LaMastro, Joey N. Upshaw, Jenica N. Pulakat, Lakshmi Jaffe, Iris Z. London, Cheryl A. Chen, Howard H. Yang, Vicky K. PLoS One Research Article Chemotherapy-induced impairment of autophagy is implicated in cardiac toxicity induced by anti-cancer drugs. Imperfect translation from rodent models and lack of in vitro models of toxicity has limited investigation of autophagic flux dysregulation, preventing design of novel cardioprotective strategies based on autophagy control. Development of an adult heart tissue culture technique from a translational model will improve investigation of cardiac toxicity. We aimed to optimize a canine cardiac slice culture system for exploration of cancer therapy impact on intact cardiac tissue, creating a translatable model that maintains autophagy in culture and is amenable to autophagy modulation. Canine cardiac tissue slices (350 μm) were generated from left ventricular free wall collected from euthanized client-owned dogs (n = 7) free of cardiovascular disease at the Foster Hospital for Small Animals at Tufts University. Cell viability and apoptosis were quantified with MTT assay and TUNEL staining. Cardiac slices were challenged with doxorubicin and an autophagy activator (rapamycin) or inhibitor (chloroquine). Autophagic flux components (LC3, p62) were quantified by western blot. Cardiac slices retained high cell viability for >7 days in culture and basal levels of autophagic markers remained unchanged. Doxorubicin treatment resulted in perturbation of the autophagic flux and cell death, while rapamycin co-treatment restored normal autophagic flux and maintained cell survival. We developed an adult canine cardiac slice culture system appropriate for studying the effects of autophagic flux that may be applicable to drug toxicity evaluations. Public Library of Science 2023-03-16 /pmc/articles/PMC10019679/ /pubmed/36928870 http://dx.doi.org/10.1371/journal.pone.0282859 Text en © 2023 Boukhalfa et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Boukhalfa, Asma Robinson, Sally R. Meola, Dawn M. Robinson, Nicholas A. Ling, Lauren A. LaMastro, Joey N. Upshaw, Jenica N. Pulakat, Lakshmi Jaffe, Iris Z. London, Cheryl A. Chen, Howard H. Yang, Vicky K. Using cultured canine cardiac slices to model the autophagic flux with doxorubicin |
title | Using cultured canine cardiac slices to model the autophagic flux with doxorubicin |
title_full | Using cultured canine cardiac slices to model the autophagic flux with doxorubicin |
title_fullStr | Using cultured canine cardiac slices to model the autophagic flux with doxorubicin |
title_full_unstemmed | Using cultured canine cardiac slices to model the autophagic flux with doxorubicin |
title_short | Using cultured canine cardiac slices to model the autophagic flux with doxorubicin |
title_sort | using cultured canine cardiac slices to model the autophagic flux with doxorubicin |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10019679/ https://www.ncbi.nlm.nih.gov/pubmed/36928870 http://dx.doi.org/10.1371/journal.pone.0282859 |
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