Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune Response
Hypoxia is a feature of neurodegenerative diseases, and can both directly and indirectly impact on neuronal function through modulation of glial function. Astrocytes play a key role in regulating homeostasis within the central nervous system, and mediate hypoxia-induced changes in response to reduce...
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MDPI AG
2020-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/21/8028 |
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| author | Scott P. Allen Rajpinder Singh Seehra Paul R. Heath Benjamin P. C. Hall Jessica Bates Claire J. Garwood Martyna M. Matuszyk Stephen B. Wharton Julie E. Simpson |
| author_facet | Scott P. Allen Rajpinder Singh Seehra Paul R. Heath Benjamin P. C. Hall Jessica Bates Claire J. Garwood Martyna M. Matuszyk Stephen B. Wharton Julie E. Simpson |
| author_sort | Scott P. Allen |
| collection | DOAJ |
| description | Hypoxia is a feature of neurodegenerative diseases, and can both directly and indirectly impact on neuronal function through modulation of glial function. Astrocytes play a key role in regulating homeostasis within the central nervous system, and mediate hypoxia-induced changes in response to reduced oxygen availability. The current study performed a detailed characterization of hypoxia-induced changes in the transcriptomic profile of astrocytes in vitro. Human astrocytes were cultured under normoxic (5% CO<sub>2</sub>, 95% air) or hypoxic conditions (1% O<sub>2</sub>, 5% CO<sub>2</sub>, 94% N<sub>2</sub>) for 24 h, and the gene expression profile assessed by microarray analysis. In response to hypoxia 4904 genes were significantly differentially expressed (1306 upregulated and 3598 downregulated, FC ≥ 2 and <i>p</i> ≤ 0.05). Analysis of the significant differentially expressed transcripts identified an increase in immune response pathways, and dysregulation of signalling pathways, including HIF-1 (<i>p</i> = 0.002), and metabolism, including glycolysis (<i>p</i> = 0.006). To assess whether the hypoxia-induced metabolic gene changes observed affected metabolism at a functional level, both the glycolytic and mitochondrial flux were measured using an XF bioanalyser. In support of the transcriptomic data, under physiological conditions hypoxia significantly reduced mitochondrial respiratory flux (<i>p</i> = 0.0001) but increased basal glycolytic flux (<i>p</i> = 0.0313). However, when metabolically stressed, hypoxia reduced mitochondrial spare respiratory capacity (<i>p</i> = 0.0485) and both glycolytic capacity (<i>p</i> = 0.0001) and glycolytic reserve (<i>p</i> < 0.0001). In summary, the current findings detail hypoxia-induced changes in the astrocyte transcriptome in vitro, identifying potential targets for modifying the astrocyte response to reduced oxygen availability in pathological conditions associated with ischaemia/hypoxia, including manipulation of mitochondrial function, metabolism, and the immune response. |
| first_indexed | 2024-03-10T15:16:48Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T15:16:48Z |
| publishDate | 2020-10-01 |
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| spelling | doaj.art-58ac9fb6b2bb4dc3bb18f7fcdc0fb23d2023-11-20T18:51:42ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-10-012121802810.3390/ijms21218028Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune ResponseScott P. Allen0Rajpinder Singh Seehra1Paul R. Heath2Benjamin P. C. Hall3Jessica Bates4Claire J. Garwood5Martyna M. Matuszyk6Stephen B. Wharton7Julie E. Simpson8Sheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKSheffield Institute for Translational Neuroscience, 385a Glossop Road, University of Sheffield, Sheffield S10 2HQ, UKHypoxia is a feature of neurodegenerative diseases, and can both directly and indirectly impact on neuronal function through modulation of glial function. Astrocytes play a key role in regulating homeostasis within the central nervous system, and mediate hypoxia-induced changes in response to reduced oxygen availability. The current study performed a detailed characterization of hypoxia-induced changes in the transcriptomic profile of astrocytes in vitro. Human astrocytes were cultured under normoxic (5% CO<sub>2</sub>, 95% air) or hypoxic conditions (1% O<sub>2</sub>, 5% CO<sub>2</sub>, 94% N<sub>2</sub>) for 24 h, and the gene expression profile assessed by microarray analysis. In response to hypoxia 4904 genes were significantly differentially expressed (1306 upregulated and 3598 downregulated, FC ≥ 2 and <i>p</i> ≤ 0.05). Analysis of the significant differentially expressed transcripts identified an increase in immune response pathways, and dysregulation of signalling pathways, including HIF-1 (<i>p</i> = 0.002), and metabolism, including glycolysis (<i>p</i> = 0.006). To assess whether the hypoxia-induced metabolic gene changes observed affected metabolism at a functional level, both the glycolytic and mitochondrial flux were measured using an XF bioanalyser. In support of the transcriptomic data, under physiological conditions hypoxia significantly reduced mitochondrial respiratory flux (<i>p</i> = 0.0001) but increased basal glycolytic flux (<i>p</i> = 0.0313). However, when metabolically stressed, hypoxia reduced mitochondrial spare respiratory capacity (<i>p</i> = 0.0485) and both glycolytic capacity (<i>p</i> = 0.0001) and glycolytic reserve (<i>p</i> < 0.0001). In summary, the current findings detail hypoxia-induced changes in the astrocyte transcriptome in vitro, identifying potential targets for modifying the astrocyte response to reduced oxygen availability in pathological conditions associated with ischaemia/hypoxia, including manipulation of mitochondrial function, metabolism, and the immune response.https://www.mdpi.com/1422-0067/21/21/8028astrocytehypoxiaimmune responsemetabolismmitochondrial dysfunctiontranscriptome |
| spellingShingle | Scott P. Allen Rajpinder Singh Seehra Paul R. Heath Benjamin P. C. Hall Jessica Bates Claire J. Garwood Martyna M. Matuszyk Stephen B. Wharton Julie E. Simpson Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune Response International Journal of Molecular Sciences astrocyte hypoxia immune response metabolism mitochondrial dysfunction transcriptome |
| title | Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune Response |
| title_full | Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune Response |
| title_fullStr | Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune Response |
| title_full_unstemmed | Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune Response |
| title_short | Transcriptomic Analysis of Human Astrocytes In Vitro Reveals Hypoxia-Induced Mitochondrial Dysfunction, Modulation of Metabolism, and Dysregulation of the Immune Response |
| title_sort | transcriptomic analysis of human astrocytes in vitro reveals hypoxia induced mitochondrial dysfunction modulation of metabolism and dysregulation of the immune response |
| topic | astrocyte hypoxia immune response metabolism mitochondrial dysfunction transcriptome |
| url | https://www.mdpi.com/1422-0067/21/21/8028 |
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