Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R level
Abstract Insulin resistance impairs the cellular insulin response, and often precedes metabolic disorders, like type 2 diabetes, impacting an increasing number of people globally. Understanding the molecular mechanisms in hepatic insulin resistance is essential for early preventive treatments. To el...
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Nature Portfolio
2024-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-77817-5 |
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| author | Sarah Hyllekvist Jørgensen Kristina Bennet Emdal Anna-Kathrine Pedersen Lene Nygaard Axelsen Helene Faustrup Kildegaard Damien Demozay Thomas Åskov Pedersen Mads Grønborg Rita Slaaby Peter Kresten Nielsen Jesper Velgaard Olsen |
| author_facet | Sarah Hyllekvist Jørgensen Kristina Bennet Emdal Anna-Kathrine Pedersen Lene Nygaard Axelsen Helene Faustrup Kildegaard Damien Demozay Thomas Åskov Pedersen Mads Grønborg Rita Slaaby Peter Kresten Nielsen Jesper Velgaard Olsen |
| author_sort | Sarah Hyllekvist Jørgensen |
| collection | DOAJ |
| description | Abstract Insulin resistance impairs the cellular insulin response, and often precedes metabolic disorders, like type 2 diabetes, impacting an increasing number of people globally. Understanding the molecular mechanisms in hepatic insulin resistance is essential for early preventive treatments. To elucidate changes in insulin signal transduction associated with hepatocellular resistance, we employed a multi-layered mass spectrometry-based proteomics approach focused on insulin receptor (IR) signaling at the interactome, phosphoproteome, and proteome levels in a long-term hyperinsulinemia-induced insulin-resistant HepG2 cell line with a knockout of the insulin-like growth factor 1 receptor (IGF1R KO). The analysis revealed insulin-stimulated recruitment of the PI3K complex in both insulin-sensitive and -resistant cells. Phosphoproteomics showed attenuated signaling via the metabolic PI3K-AKT pathway but sustained extracellular signal-regulated kinase (ERK) activity in insulin-resistant cells. At the proteome level, the ephrin type-A receptor 2 (EphA2) showed an insulin-induced increase in expression, which occurred through the ERK signaling pathway and was concordantly independent of insulin resistance. Induction of EphA2 by insulin was confirmed in additional cell lines and observed uniquely in cells with high IR-to-IGF1R ratio. The multi-layered proteomics dataset provided insights into insulin signaling, serving as a resource to generate and test hypotheses, leading to an improved understanding of insulin resistance. |
| format | Article |
| id | doaj-art-78e6b6a506d841aca705b82fea36c55f |
| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-78e6b6a506d841aca705b82fea36c55f2025-08-20T02:22:24ZengNature PortfolioScientific Reports2045-23222024-11-0114112010.1038/s41598-024-77817-5Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R levelSarah Hyllekvist Jørgensen0Kristina Bennet Emdal1Anna-Kathrine Pedersen2Lene Nygaard Axelsen3Helene Faustrup Kildegaard4Damien Demozay5Thomas Åskov Pedersen6Mads Grønborg7Rita Slaaby8Peter Kresten Nielsen9Jesper Velgaard Olsen10Proteomics Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenProteomics Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenProteomics Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenGlobal Translation, Novo Nordisk A/SGlobal Nucleic Acid Therapies, Novo Nordisk A/SGlobal Drug Discovery, Novo Nordisk A/SGlobal Drug Discovery, Novo Nordisk A/SGlobal Translation, Novo Nordisk A/SGlobal Drug Discovery, Novo Nordisk A/SGlobal Research Technologies, Novo Nordisk A/SProteomics Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of CopenhagenAbstract Insulin resistance impairs the cellular insulin response, and often precedes metabolic disorders, like type 2 diabetes, impacting an increasing number of people globally. Understanding the molecular mechanisms in hepatic insulin resistance is essential for early preventive treatments. To elucidate changes in insulin signal transduction associated with hepatocellular resistance, we employed a multi-layered mass spectrometry-based proteomics approach focused on insulin receptor (IR) signaling at the interactome, phosphoproteome, and proteome levels in a long-term hyperinsulinemia-induced insulin-resistant HepG2 cell line with a knockout of the insulin-like growth factor 1 receptor (IGF1R KO). The analysis revealed insulin-stimulated recruitment of the PI3K complex in both insulin-sensitive and -resistant cells. Phosphoproteomics showed attenuated signaling via the metabolic PI3K-AKT pathway but sustained extracellular signal-regulated kinase (ERK) activity in insulin-resistant cells. At the proteome level, the ephrin type-A receptor 2 (EphA2) showed an insulin-induced increase in expression, which occurred through the ERK signaling pathway and was concordantly independent of insulin resistance. Induction of EphA2 by insulin was confirmed in additional cell lines and observed uniquely in cells with high IR-to-IGF1R ratio. The multi-layered proteomics dataset provided insights into insulin signaling, serving as a resource to generate and test hypotheses, leading to an improved understanding of insulin resistance.https://doi.org/10.1038/s41598-024-77817-5 |
| spellingShingle | Sarah Hyllekvist Jørgensen Kristina Bennet Emdal Anna-Kathrine Pedersen Lene Nygaard Axelsen Helene Faustrup Kildegaard Damien Demozay Thomas Åskov Pedersen Mads Grønborg Rita Slaaby Peter Kresten Nielsen Jesper Velgaard Olsen Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R level Scientific Reports |
| title | Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R level |
| title_full | Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R level |
| title_fullStr | Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R level |
| title_full_unstemmed | Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R level |
| title_short | Multi-layered proteomics identifies insulin-induced upregulation of the EphA2 receptor via the ERK pathway which is dependent on low IGF1R level |
| title_sort | multi layered proteomics identifies insulin induced upregulation of the epha2 receptor via the erk pathway which is dependent on low igf1r level |
| url | https://doi.org/10.1038/s41598-024-77817-5 |
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