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The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes

Population data have shown an association between higher circulating branched-chain amino acids (BCAA) and the severity of insulin resistance in people with diabetes. While several studies have assessed BCAA metabolism as a potential target for regulation, less attention has been paid to the role of...

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Autores principales: Rivera, Caroline N., Smith, Carly E., Draper, Lillian V., Ochoa, Gabriela E., Watne, Rachel M., Wommack, Andrew J., Vaughan, Roger A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10303577/
https://www.ncbi.nlm.nih.gov/pubmed/37367923
http://dx.doi.org/10.3390/metabo13060766
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author Rivera, Caroline N.
Smith, Carly E.
Draper, Lillian V.
Ochoa, Gabriela E.
Watne, Rachel M.
Wommack, Andrew J.
Vaughan, Roger A.
author_facet Rivera, Caroline N.
Smith, Carly E.
Draper, Lillian V.
Ochoa, Gabriela E.
Watne, Rachel M.
Wommack, Andrew J.
Vaughan, Roger A.
author_sort Rivera, Caroline N.
collection PubMed
description Population data have shown an association between higher circulating branched-chain amino acids (BCAA) and the severity of insulin resistance in people with diabetes. While several studies have assessed BCAA metabolism as a potential target for regulation, less attention has been paid to the role of L-type amino acid transporter 1 (LAT1), the primary transporter of BCAA in skeletal muscle. The aim of this study was to assess the impact of JPH203 (JPH), a LAT1 inhibitor, on myotube metabolism in both insulin-sensitive and insulin-resistant myotubes. C2C12 myotubes were treated with or without 1 μM or 2 μM JPH for 24 h with or without insulin resistance. Western blot and qRT-PCR were used to assess protein content and gene expression, respectively. Mitochondrial and glycolytic metabolism were measured via Seahorse Assay, and fluorescent staining was used to measure mitochondrial content. BCAA media content was quantified using liquid chromatography–mass spectrometry. JPH at 1 μM (but not 2 μM) increased mitochondrial metabolism and content without inducing changes in mRNA expression of transcripts associated with mitochondrial biogenesis or mitochondrial dynamics. Along with increased mitochondrial function, 1μM treatment also reduced extracellular leucine and valine. JPH at 2 μM reduced pAkt signaling and increased extracellular accumulation of isoleucine without inducing changes in BCAA metabolic genes. Collectively, JPH may increase mitochondrial function independent of the mitochondrial biogenic transcription pathway; however, high doses may reduce insulin signaling.
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spelling pubmed-103035772023-06-29 The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes Rivera, Caroline N. Smith, Carly E. Draper, Lillian V. Ochoa, Gabriela E. Watne, Rachel M. Wommack, Andrew J. Vaughan, Roger A. Metabolites Article Population data have shown an association between higher circulating branched-chain amino acids (BCAA) and the severity of insulin resistance in people with diabetes. While several studies have assessed BCAA metabolism as a potential target for regulation, less attention has been paid to the role of L-type amino acid transporter 1 (LAT1), the primary transporter of BCAA in skeletal muscle. The aim of this study was to assess the impact of JPH203 (JPH), a LAT1 inhibitor, on myotube metabolism in both insulin-sensitive and insulin-resistant myotubes. C2C12 myotubes were treated with or without 1 μM or 2 μM JPH for 24 h with or without insulin resistance. Western blot and qRT-PCR were used to assess protein content and gene expression, respectively. Mitochondrial and glycolytic metabolism were measured via Seahorse Assay, and fluorescent staining was used to measure mitochondrial content. BCAA media content was quantified using liquid chromatography–mass spectrometry. JPH at 1 μM (but not 2 μM) increased mitochondrial metabolism and content without inducing changes in mRNA expression of transcripts associated with mitochondrial biogenesis or mitochondrial dynamics. Along with increased mitochondrial function, 1μM treatment also reduced extracellular leucine and valine. JPH at 2 μM reduced pAkt signaling and increased extracellular accumulation of isoleucine without inducing changes in BCAA metabolic genes. Collectively, JPH may increase mitochondrial function independent of the mitochondrial biogenic transcription pathway; however, high doses may reduce insulin signaling. MDPI 2023-06-19 /pmc/articles/PMC10303577/ /pubmed/37367923 http://dx.doi.org/10.3390/metabo13060766 Text en © 2023 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
Rivera, Caroline N.
Smith, Carly E.
Draper, Lillian V.
Ochoa, Gabriela E.
Watne, Rachel M.
Wommack, Andrew J.
Vaughan, Roger A.
The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes
title The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes
title_full The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes
title_fullStr The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes
title_full_unstemmed The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes
title_short The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes
title_sort selective lat1 inhibitor jph203 enhances mitochondrial metabolism and content in insulin-sensitive and insulin-resistant c2c12 myotubes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10303577/
https://www.ncbi.nlm.nih.gov/pubmed/37367923
http://dx.doi.org/10.3390/metabo13060766
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