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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
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.
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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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