Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effect
The small-molecule inhibitor of phosphoglycerate dehydrogenase, NCT-503, reduces incorporation of glucose-derived carbons into serine in vitro. Here we describe an off-target effect of NCT-503 in neuroblastoma cell lines expressing divergent phosphoglycerate dehydrogenase (PHGDH) levels and single-c...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2021-01-01
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| Series: | Journal of Enzyme Inhibition and Medicinal Chemistry |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/14756366.2021.1935917 |
| _version_ | 1798016100402724864 |
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| author | Birte Arlt Guido Mastrobuoni Jasmin Wuenschel Kathy Astrahantseff Angelika Eggert Stefan Kempa Hedwig E. Deubzer |
| author_facet | Birte Arlt Guido Mastrobuoni Jasmin Wuenschel Kathy Astrahantseff Angelika Eggert Stefan Kempa Hedwig E. Deubzer |
| author_sort | Birte Arlt |
| collection | DOAJ |
| description | The small-molecule inhibitor of phosphoglycerate dehydrogenase, NCT-503, reduces incorporation of glucose-derived carbons into serine in vitro. Here we describe an off-target effect of NCT-503 in neuroblastoma cell lines expressing divergent phosphoglycerate dehydrogenase (PHGDH) levels and single-cell clones with CRISPR-Cas9-directed PHGDH knockout or their respective wildtype controls. NCT-503 treatment strongly reduced synthesis of glucose-derived citrate in all cell models investigated compared to the inactive drug control and independent of PHGDH expression level. Incorporation of glucose-derived carbons entering the TCA cycle via pyruvate carboxylase was enhanced by NCT-503 treatment. The activity of citrate synthase was not altered by NCT-503 treatment. We also detected no change in the thermal stabilisation of citrate synthase in cellular thermal shift assays from NCT-503-treated cells. Thus, the direct cause of the observed off-target effect remains enigmatic. Our findings highlight off-target potential within a metabolic assessment of carbon usage in cells treated with the small-molecule inhibitor, NCT-503. |
| first_indexed | 2024-04-11T15:44:16Z |
| format | Article |
| id | doaj.art-0116dd5b878942ba90fb3c088f812553 |
| institution | Directory Open Access Journal |
| issn | 1475-6366 1475-6374 |
| language | English |
| last_indexed | 2024-04-11T15:44:16Z |
| publishDate | 2021-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Journal of Enzyme Inhibition and Medicinal Chemistry |
| spelling | doaj.art-0116dd5b878942ba90fb3c088f8125532022-12-22T04:15:37ZengTaylor & Francis GroupJournal of Enzyme Inhibition and Medicinal Chemistry1475-63661475-63742021-01-013611282128910.1080/14756366.2021.19359171935917Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effectBirte Arlt0Guido Mastrobuoni1Jasmin Wuenschel2Kathy Astrahantseff3Angelika Eggert4Stefan Kempa5Hedwig E. Deubzer6Department of Pediatric Hematology and Oncology, Charité – Universitätsmedizin BerlinIntegrative Proteomics and Metabolomics, Berlin Institute for Medical Systems Biology at the Max-Delbrück Center for Molecular Medicine in the Helmholtz AssociationDepartment of Pediatric Hematology and Oncology, Charité – Universitätsmedizin BerlinDepartment of Pediatric Hematology and Oncology, Charité – Universitätsmedizin BerlinDepartment of Pediatric Hematology and Oncology, Charité – Universitätsmedizin BerlinBerlin Institute of Health (BIH)Department of Pediatric Hematology and Oncology, Charité – Universitätsmedizin BerlinThe small-molecule inhibitor of phosphoglycerate dehydrogenase, NCT-503, reduces incorporation of glucose-derived carbons into serine in vitro. Here we describe an off-target effect of NCT-503 in neuroblastoma cell lines expressing divergent phosphoglycerate dehydrogenase (PHGDH) levels and single-cell clones with CRISPR-Cas9-directed PHGDH knockout or their respective wildtype controls. NCT-503 treatment strongly reduced synthesis of glucose-derived citrate in all cell models investigated compared to the inactive drug control and independent of PHGDH expression level. Incorporation of glucose-derived carbons entering the TCA cycle via pyruvate carboxylase was enhanced by NCT-503 treatment. The activity of citrate synthase was not altered by NCT-503 treatment. We also detected no change in the thermal stabilisation of citrate synthase in cellular thermal shift assays from NCT-503-treated cells. Thus, the direct cause of the observed off-target effect remains enigmatic. Our findings highlight off-target potential within a metabolic assessment of carbon usage in cells treated with the small-molecule inhibitor, NCT-503.http://dx.doi.org/10.1080/14756366.2021.1935917cancer cell metabolismde novo serine synthesis pathwaycitrate synthasepulsed stable isotope-resolved metabolomicsthermal shift assay |
| spellingShingle | Birte Arlt Guido Mastrobuoni Jasmin Wuenschel Kathy Astrahantseff Angelika Eggert Stefan Kempa Hedwig E. Deubzer Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effect Journal of Enzyme Inhibition and Medicinal Chemistry cancer cell metabolism de novo serine synthesis pathway citrate synthase pulsed stable isotope-resolved metabolomics thermal shift assay |
| title | Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effect |
| title_full | Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effect |
| title_fullStr | Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effect |
| title_full_unstemmed | Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effect |
| title_short | Inhibiting PHGDH with NCT-503 reroutes glucose-derived carbons into the TCA cycle, independently of its on-target effect |
| title_sort | inhibiting phgdh with nct 503 reroutes glucose derived carbons into the tca cycle independently of its on target effect |
| topic | cancer cell metabolism de novo serine synthesis pathway citrate synthase pulsed stable isotope-resolved metabolomics thermal shift assay |
| url | http://dx.doi.org/10.1080/14756366.2021.1935917 |
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