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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MDPI AG
2023-06-01
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| Series: | Metabolites |
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| Online Access: | https://www.mdpi.com/2218-1989/13/6/766 |
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| author | Caroline N. Rivera Carly E. Smith Lillian V. Draper Gabriela E. Ochoa Rachel M. Watne Andrew J. Wommack Roger A. Vaughan |
| author_facet | Caroline N. Rivera Carly E. Smith Lillian V. Draper Gabriela E. Ochoa Rachel M. Watne Andrew J. Wommack Roger A. Vaughan |
| author_sort | Caroline N. Rivera |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-11T02:10:24Z |
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| id | doaj.art-6c78959572fb4c7f8df9d6202467c745 |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-11T02:10:24Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-6c78959572fb4c7f8df9d6202467c7452023-11-18T11:35:23ZengMDPI AGMetabolites2218-19892023-06-0113676610.3390/metabo13060766The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 MyotubesCaroline N. Rivera0Carly E. Smith1Lillian V. Draper2Gabriela E. Ochoa3Rachel M. Watne4Andrew J. Wommack5Roger A. Vaughan6Department of Exercise Science, High Point University, High Point, NC 27268, USADepartment of Exercise Science, High Point University, High Point, NC 27268, USADepartment of Exercise Science, High Point University, High Point, NC 27268, USADepartment of Exercise Science, High Point University, High Point, NC 27268, USADepartment of Chemistry, High Point University, High Point, NC 27268, USADepartment of Chemistry, High Point University, High Point, NC 27268, USADepartment of Exercise Science, High Point University, High Point, NC 27268, USAPopulation 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.https://www.mdpi.com/2218-1989/13/6/766leucineisoleucinevalinepAkt/Aktskeletal muscleinsulin resistance |
| spellingShingle | Caroline N. Rivera Carly E. Smith Lillian V. Draper Gabriela E. Ochoa Rachel M. Watne Andrew J. Wommack Roger A. Vaughan The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes Metabolites leucine isoleucine valine pAkt/Akt skeletal muscle insulin resistance |
| 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 | leucine isoleucine valine pAkt/Akt skeletal muscle insulin resistance |
| url | https://www.mdpi.com/2218-1989/13/6/766 |
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