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Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity
Nephrotoxicity is a major cause of kidney disease and failure in drug development, but understanding of cellular mechanisms is limited, highlighting the need for better experimental models and methodological approaches. Most nephrotoxins damage the proximal tubule (PT), causing functional impairment...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9840465/ https://www.ncbi.nlm.nih.gov/pubmed/36654930 http://dx.doi.org/10.1093/function/zqac065 |
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author | Pearson, Adam Haenni, Dominik Bouitbir, Jamal Hunt, Matthew Payne, Brendan A I Sachdeva, Ashwin Hung, Rachel K Y Post, Frank A Connolly, John Nlandu-Khodo, Stellor Jankovic, Nevena Bugarski, Milica Hall, Andrew M |
author_facet | Pearson, Adam Haenni, Dominik Bouitbir, Jamal Hunt, Matthew Payne, Brendan A I Sachdeva, Ashwin Hung, Rachel K Y Post, Frank A Connolly, John Nlandu-Khodo, Stellor Jankovic, Nevena Bugarski, Milica Hall, Andrew M |
author_sort | Pearson, Adam |
collection | PubMed |
description | Nephrotoxicity is a major cause of kidney disease and failure in drug development, but understanding of cellular mechanisms is limited, highlighting the need for better experimental models and methodological approaches. Most nephrotoxins damage the proximal tubule (PT), causing functional impairment of solute reabsorption and systemic metabolic complications. The antiviral drug tenofovir disoproxil fumarate (TDF) is an archetypal nephrotoxin, inducing mitochondrial abnormalities and urinary solute wasting, for reasons that were previously unclear. Here, we developed an automated, high-throughput imaging pipeline to screen the effects of TDF on solute transport and mitochondrial morphology in human-derived RPTEC/TERT1 cells, and leveraged this to generate realistic models of functional toxicity. By applying multiparametric metabolic profiling—including oxygen consumption measurements, metabolomics, and transcriptomics—we elucidated a highly robust molecular fingerprint of TDF exposure. Crucially, we identified that the active metabolite inhibits complex V (ATP synthase), and that TDF treatment causes rapid, dose-dependent loss of complex V activity and expression. Moreover, we found evidence of complex V suppression in kidney biopsies from humans with TDF toxicity. Thus, we demonstrate an effective and convenient experimental approach to screen for disease relevant functional defects in kidney cells in vitro, and reveal a new paradigm for understanding the pathogenesis of a substantial cause of nephrotoxicity. |
format | Online Article Text |
id | pubmed-9840465 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-98404652023-01-17 Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity Pearson, Adam Haenni, Dominik Bouitbir, Jamal Hunt, Matthew Payne, Brendan A I Sachdeva, Ashwin Hung, Rachel K Y Post, Frank A Connolly, John Nlandu-Khodo, Stellor Jankovic, Nevena Bugarski, Milica Hall, Andrew M Function (Oxf) Research Article Nephrotoxicity is a major cause of kidney disease and failure in drug development, but understanding of cellular mechanisms is limited, highlighting the need for better experimental models and methodological approaches. Most nephrotoxins damage the proximal tubule (PT), causing functional impairment of solute reabsorption and systemic metabolic complications. The antiviral drug tenofovir disoproxil fumarate (TDF) is an archetypal nephrotoxin, inducing mitochondrial abnormalities and urinary solute wasting, for reasons that were previously unclear. Here, we developed an automated, high-throughput imaging pipeline to screen the effects of TDF on solute transport and mitochondrial morphology in human-derived RPTEC/TERT1 cells, and leveraged this to generate realistic models of functional toxicity. By applying multiparametric metabolic profiling—including oxygen consumption measurements, metabolomics, and transcriptomics—we elucidated a highly robust molecular fingerprint of TDF exposure. Crucially, we identified that the active metabolite inhibits complex V (ATP synthase), and that TDF treatment causes rapid, dose-dependent loss of complex V activity and expression. Moreover, we found evidence of complex V suppression in kidney biopsies from humans with TDF toxicity. Thus, we demonstrate an effective and convenient experimental approach to screen for disease relevant functional defects in kidney cells in vitro, and reveal a new paradigm for understanding the pathogenesis of a substantial cause of nephrotoxicity. Oxford University Press 2022-12-24 /pmc/articles/PMC9840465/ /pubmed/36654930 http://dx.doi.org/10.1093/function/zqac065 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of American Physiological Society. 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 reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Pearson, Adam Haenni, Dominik Bouitbir, Jamal Hunt, Matthew Payne, Brendan A I Sachdeva, Ashwin Hung, Rachel K Y Post, Frank A Connolly, John Nlandu-Khodo, Stellor Jankovic, Nevena Bugarski, Milica Hall, Andrew M Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity |
title | Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity |
title_full | Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity |
title_fullStr | Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity |
title_full_unstemmed | Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity |
title_short | Integration of High-Throughput Imaging and Multiparametric Metabolic Profiling Reveals a Mitochondrial Mechanism of Tenofovir Toxicity |
title_sort | integration of high-throughput imaging and multiparametric metabolic profiling reveals a mitochondrial mechanism of tenofovir toxicity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9840465/ https://www.ncbi.nlm.nih.gov/pubmed/36654930 http://dx.doi.org/10.1093/function/zqac065 |
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