<sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA Activity
Disrupted endothelial metabolism is linked to endothelial dysfunction and cardiovascular disease. Targeted metabolic inhibitors are potential therapeutics; however, their systemic impact on endothelial metabolism remains unknown. In this study, we combined stable isotope labeling with <sup>13&...
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MDPI AG
2021-04-01
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| Online Access: | https://www.mdpi.com/2218-1989/11/4/226 |
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| author | Bilal Moiz Jonathan Garcia Sarah Basehore Angela Sun Andrew Li Surya Padmanabhan Kaitlyn Albus Cholsoon Jang Ganesh Sriram Alisa Morss Clyne |
| author_facet | Bilal Moiz Jonathan Garcia Sarah Basehore Angela Sun Andrew Li Surya Padmanabhan Kaitlyn Albus Cholsoon Jang Ganesh Sriram Alisa Morss Clyne |
| author_sort | Bilal Moiz |
| collection | DOAJ |
| description | Disrupted endothelial metabolism is linked to endothelial dysfunction and cardiovascular disease. Targeted metabolic inhibitors are potential therapeutics; however, their systemic impact on endothelial metabolism remains unknown. In this study, we combined stable isotope labeling with <sup>13</sup>C metabolic flux analysis (<sup>13</sup>C MFA) to determine how targeted inhibition of the polyol (fidarestat), pentose phosphate (DHEA), and hexosamine biosynthetic (azaserine) pathways alters endothelial metabolism. Glucose, glutamine, and a four-carbon input to the malate shuttle were important carbon sources in the baseline human umbilical vein endothelial cell (HUVEC) <sup>13</sup>C MFA model. We observed two to three times higher glutamine uptake in fidarestat and azaserine-treated cells. Fidarestat and DHEA-treated HUVEC showed decreased <sup>13</sup>C enrichment of glycolytic and TCA metabolites and amino acids. Azaserine-treated HUVEC primarily showed <sup>13</sup>C enrichment differences in UDP-GlcNAc. <sup>13</sup>C MFA estimated decreased pentose phosphate pathway flux and increased TCA activity with reversed malate shuttle direction in fidarestat and DHEA-treated HUVEC. In contrast, <sup>13</sup>C MFA estimated increases in both pentose phosphate pathway and TCA activity in azaserine-treated cells. These data show the potential importance of endothelial malate shuttle activity and suggest that inhibiting glycolytic side branch pathways can change the metabolic network, highlighting the need to study systemic metabolic therapeutic effects. |
| first_indexed | 2024-03-10T12:31:45Z |
| format | Article |
| id | doaj.art-43b11d1840e3462cbebeb1caab961d74 |
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| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-10T12:31:45Z |
| publishDate | 2021-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-43b11d1840e3462cbebeb1caab961d742023-11-21T14:35:11ZengMDPI AGMetabolites2218-19892021-04-0111422610.3390/metabo11040226<sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA ActivityBilal Moiz0Jonathan Garcia1Sarah Basehore2Angela Sun3Andrew Li4Surya Padmanabhan5Kaitlyn Albus6Cholsoon Jang7Ganesh Sriram8Alisa Morss Clyne9Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USASchool of Bioengineering, Science, and Heath Systems, Drexel University, Philadelphia, PA 19104, USASchool of Bioengineering, Science, and Heath Systems, Drexel University, Philadelphia, PA 19104, USAFischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USAFischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USAFischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USAFischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USADepartment of Biological Chemistry, Chao Family Comprehensive Cancer Center, University of California Irvine, Irvine, CA 92697, USADepartment of Chemical and Biomolecular Engineering, University of Maryland, College Park, MD 20742, USAFischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USADisrupted endothelial metabolism is linked to endothelial dysfunction and cardiovascular disease. Targeted metabolic inhibitors are potential therapeutics; however, their systemic impact on endothelial metabolism remains unknown. In this study, we combined stable isotope labeling with <sup>13</sup>C metabolic flux analysis (<sup>13</sup>C MFA) to determine how targeted inhibition of the polyol (fidarestat), pentose phosphate (DHEA), and hexosamine biosynthetic (azaserine) pathways alters endothelial metabolism. Glucose, glutamine, and a four-carbon input to the malate shuttle were important carbon sources in the baseline human umbilical vein endothelial cell (HUVEC) <sup>13</sup>C MFA model. We observed two to three times higher glutamine uptake in fidarestat and azaserine-treated cells. Fidarestat and DHEA-treated HUVEC showed decreased <sup>13</sup>C enrichment of glycolytic and TCA metabolites and amino acids. Azaserine-treated HUVEC primarily showed <sup>13</sup>C enrichment differences in UDP-GlcNAc. <sup>13</sup>C MFA estimated decreased pentose phosphate pathway flux and increased TCA activity with reversed malate shuttle direction in fidarestat and DHEA-treated HUVEC. In contrast, <sup>13</sup>C MFA estimated increases in both pentose phosphate pathway and TCA activity in azaserine-treated cells. These data show the potential importance of endothelial malate shuttle activity and suggest that inhibiting glycolytic side branch pathways can change the metabolic network, highlighting the need to study systemic metabolic therapeutic effects.https://www.mdpi.com/2218-1989/11/4/226metabolic flux analysisfluxomicsendothelial metabolismcardiovascular diseasepolyol pathwaypentose phosphate pathway |
| spellingShingle | Bilal Moiz Jonathan Garcia Sarah Basehore Angela Sun Andrew Li Surya Padmanabhan Kaitlyn Albus Cholsoon Jang Ganesh Sriram Alisa Morss Clyne <sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA Activity Metabolites metabolic flux analysis fluxomics endothelial metabolism cardiovascular disease polyol pathway pentose phosphate pathway |
| title | <sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA Activity |
| title_full | <sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA Activity |
| title_fullStr | <sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA Activity |
| title_full_unstemmed | <sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA Activity |
| title_short | <sup>13</sup>C Metabolic Flux Analysis Indicates Endothelial Cells Attenuate Metabolic Perturbations by Modulating TCA Activity |
| title_sort | sup 13 sup c metabolic flux analysis indicates endothelial cells attenuate metabolic perturbations by modulating tca activity |
| topic | metabolic flux analysis fluxomics endothelial metabolism cardiovascular disease polyol pathway pentose phosphate pathway |
| url | https://www.mdpi.com/2218-1989/11/4/226 |
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