Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate
Abstract Background Extracellular vesicles (EVs) are important in the intercellular communication of the central nervous system, and their release is increased during neuroinflammation. Our previous data demonstrated an increased release of EVs during HIV-1 infection and immune activation in glial c...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2018-03-01
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| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12974-018-1120-x |
| _version_ | 1818319887350628352 |
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| author | Beiqing Wu Jianhui Liu Runze Zhao Yuju Li Justin Peer Alexander L. Braun Lixia Zhao Yi Wang Zenghan Tong Yunlong Huang Jialin C. Zheng |
| author_facet | Beiqing Wu Jianhui Liu Runze Zhao Yuju Li Justin Peer Alexander L. Braun Lixia Zhao Yi Wang Zenghan Tong Yunlong Huang Jialin C. Zheng |
| author_sort | Beiqing Wu |
| collection | DOAJ |
| description | Abstract Background Extracellular vesicles (EVs) are important in the intercellular communication of the central nervous system, and their release is increased during neuroinflammation. Our previous data demonstrated an increased release of EVs during HIV-1 infection and immune activation in glial cells. However, the molecular mechanism by which infection and inflammation increase EV release remains unknown. In the current study, we investigated the role of glutaminase 1 (GLS1)-mediated glutaminolysis and the production of a key metabolic intermediate α-ketoglutarate on EV release. Methods Human monocyte-derived macrophage primary cultures and a BV2 microglia cell line were used to represent the innate immune cells in the CNS. Transmission electron microscopy, nanoparticle tracking analysis, and Western blots were used to determine the EV regulation. GLS1 overexpression was performed using an adenovirus vector in vitro and transgenic mouse models in vivo. Data were evaluated statistically by ANOVA, followed by the Bonferroni post-test for paired observations. Results Our data revealed an increased release of EVs in GLS1-overexpressing HeLa cells. In HIV-1-infected macrophages and immune-activated microglia BV2 cells, treatment with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl)ethyl sulfide (BPTES) or CB839, two specific GLS inhibitors, significantly decreased EV release, suggesting a critical role of GLS1 in EV release. Furthermore, addition of α-ketoglutarate or ceramide rescued EV release during BPTES treatment, implicating α-ketoglutarate and ceramide as critical downstream effectors for GLS inhibitors. These findings were further corroborated with the investigation of brain tissues in GLS1-transgenic mice. The EV levels were significantly higher in GLS1 transgenic mice than those in control mice, suggesting that GLS1 increases EV release in vivo. Conclusions These findings suggest that GLS1-mediated glutaminolysis and its downstream production of α-ketoglutarate are essential in regulating EV release during HIV-1 infection and immune activation. These new mechanistic regulations may help understand how glutamine metabolism shapes EV biogenesis and release during neuroinflammation. |
| first_indexed | 2024-12-13T10:16:15Z |
| format | Article |
| id | doaj.art-3bf19a80dc62422fa1ad2a009938abbf |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-12-13T10:16:15Z |
| publishDate | 2018-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-3bf19a80dc62422fa1ad2a009938abbf2022-12-21T23:51:17ZengBMCJournal of Neuroinflammation1742-20942018-03-0115111410.1186/s12974-018-1120-xGlutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarateBeiqing Wu0Jianhui Liu1Runze Zhao2Yuju Li3Justin Peer4Alexander L. Braun5Lixia Zhao6Yi Wang7Zenghan Tong8Yunlong Huang9Jialin C. Zheng10Center for Translational Neurodegeneration and Regenerative Therapy, Shanghai Tenth People’s Hospital affiliated to Tongji University School of MedicineDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical CenterDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical CenterCenter for Translational Neurodegeneration and Regenerative Therapy, Shanghai Tenth People’s Hospital affiliated to Tongji University School of MedicineDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical CenterDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical CenterDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical CenterDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical CenterDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical CenterCenter for Translational Neurodegeneration and Regenerative Therapy, Shanghai Tenth People’s Hospital affiliated to Tongji University School of MedicineCenter for Translational Neurodegeneration and Regenerative Therapy, Shanghai Tenth People’s Hospital affiliated to Tongji University School of MedicineAbstract Background Extracellular vesicles (EVs) are important in the intercellular communication of the central nervous system, and their release is increased during neuroinflammation. Our previous data demonstrated an increased release of EVs during HIV-1 infection and immune activation in glial cells. However, the molecular mechanism by which infection and inflammation increase EV release remains unknown. In the current study, we investigated the role of glutaminase 1 (GLS1)-mediated glutaminolysis and the production of a key metabolic intermediate α-ketoglutarate on EV release. Methods Human monocyte-derived macrophage primary cultures and a BV2 microglia cell line were used to represent the innate immune cells in the CNS. Transmission electron microscopy, nanoparticle tracking analysis, and Western blots were used to determine the EV regulation. GLS1 overexpression was performed using an adenovirus vector in vitro and transgenic mouse models in vivo. Data were evaluated statistically by ANOVA, followed by the Bonferroni post-test for paired observations. Results Our data revealed an increased release of EVs in GLS1-overexpressing HeLa cells. In HIV-1-infected macrophages and immune-activated microglia BV2 cells, treatment with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl)ethyl sulfide (BPTES) or CB839, two specific GLS inhibitors, significantly decreased EV release, suggesting a critical role of GLS1 in EV release. Furthermore, addition of α-ketoglutarate or ceramide rescued EV release during BPTES treatment, implicating α-ketoglutarate and ceramide as critical downstream effectors for GLS inhibitors. These findings were further corroborated with the investigation of brain tissues in GLS1-transgenic mice. The EV levels were significantly higher in GLS1 transgenic mice than those in control mice, suggesting that GLS1 increases EV release in vivo. Conclusions These findings suggest that GLS1-mediated glutaminolysis and its downstream production of α-ketoglutarate are essential in regulating EV release during HIV-1 infection and immune activation. These new mechanistic regulations may help understand how glutamine metabolism shapes EV biogenesis and release during neuroinflammation.http://link.springer.com/article/10.1186/s12974-018-1120-xGlutamine metabolismα-KetoglutarateExtracellular vesiclesHIV-1Inflammation |
| spellingShingle | Beiqing Wu Jianhui Liu Runze Zhao Yuju Li Justin Peer Alexander L. Braun Lixia Zhao Yi Wang Zenghan Tong Yunlong Huang Jialin C. Zheng Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate Journal of Neuroinflammation Glutamine metabolism α-Ketoglutarate Extracellular vesicles HIV-1 Inflammation |
| title | Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate |
| title_full | Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate |
| title_fullStr | Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate |
| title_full_unstemmed | Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate |
| title_short | Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate |
| title_sort | glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha ketoglutarate |
| topic | Glutamine metabolism α-Ketoglutarate Extracellular vesicles HIV-1 Inflammation |
| url | http://link.springer.com/article/10.1186/s12974-018-1120-x |
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