Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells
Niemann Pick Disease Type C (NP-C), a rare neurogenetic disease with no known cure, is caused by mutations in the cholesterol trafficking protein NPC1. Brain microvascular endothelial cells (BMEC) are thought to play a critical role in the pathogenesis of several neurodegenerative diseases; however,...
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| Language: | English |
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Elsevier
2025-01-01
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| Series: | Neurobiology of Disease |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996124003711 |
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| author | Bilal Moiz Matthew Walls Viviana Alpizar Vargas Anirudh Addepalli Callie Weber Andrew Li Ganesh Sriram Alisa Morss Clyne |
| author_facet | Bilal Moiz Matthew Walls Viviana Alpizar Vargas Anirudh Addepalli Callie Weber Andrew Li Ganesh Sriram Alisa Morss Clyne |
| author_sort | Bilal Moiz |
| collection | DOAJ |
| description | Niemann Pick Disease Type C (NP-C), a rare neurogenetic disease with no known cure, is caused by mutations in the cholesterol trafficking protein NPC1. Brain microvascular endothelial cells (BMEC) are thought to play a critical role in the pathogenesis of several neurodegenerative diseases; however, little is known about how these cells are altered in NP-C. In this study, we investigated how NPC1 inhibition perturbs BMEC metabolism in human induced pluripotent stem cell-derived BMEC (hiBMEC). We incorporated extracellular metabolite and isotope labeling data into an instationary metabolic flux analysis (INST-MFA) model to estimate intracellular metabolic fluxes. We found that NPC1 inhibition significantly increased glycolysis and pentose phosphate pathway flux while decreasing mitochondrial metabolism. These changes may have been driven by gene expression changes due to increased cholesterol biosynthesis, in addition to mitochondrial cholesterol accumulation. We corroborated these findings in primary BMEC, an alternative in vitro human brain endothelial model. Finally, we found that co-treatment with hydroxypropyl-β cyclodextrin (HPβCD) partially restored metabolic phenotype in U18666A-treated BMECs, suggesting that this drug may have therapeutic effects on the brain endothelium in NP-C. Together, our data highlight the importance of NPC1 in BMEC metabolism and implicate brain endothelial dysfunction in NP-C pathogenesis. |
| format | Article |
| id | doaj-art-b4efa673ac9e4a228f22564a74bde80c |
| institution | Kabale University |
| issn | 1095-953X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj-art-b4efa673ac9e4a228f22564a74bde80c2025-01-07T04:17:09ZengElsevierNeurobiology of Disease1095-953X2025-01-01204106769Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cellsBilal Moiz0Matthew Walls1Viviana Alpizar Vargas2Anirudh Addepalli3Callie Weber4Andrew Li5Ganesh Sriram6Alisa Morss Clyne7Department of Bioengineering, University of Maryland, College Park, MD 20742, United States of AmericaDepartment of Bioengineering, University of Maryland, College Park, MD 20742, United States of AmericaDepartment of Bioengineering, University of Maryland, College Park, MD 20742, United States of AmericaDepartment of Bioengineering, University of Maryland, College Park, MD 20742, United States of AmericaDepartment of Bioengineering, University of Maryland, College Park, MD 20742, United States of AmericaDepartment of Bioengineering, University of Maryland, College Park, MD 20742, United States of AmericaDepartment of Chemical and Biochemical Engineering, University of Maryland, College Park, MD 20742, United States of AmericaDepartment of Bioengineering, University of Maryland, College Park, MD 20742, United States of America; Corresponding author at: 8278 Paint Branch Dr., College Park, MD 20742, United States of America.Niemann Pick Disease Type C (NP-C), a rare neurogenetic disease with no known cure, is caused by mutations in the cholesterol trafficking protein NPC1. Brain microvascular endothelial cells (BMEC) are thought to play a critical role in the pathogenesis of several neurodegenerative diseases; however, little is known about how these cells are altered in NP-C. In this study, we investigated how NPC1 inhibition perturbs BMEC metabolism in human induced pluripotent stem cell-derived BMEC (hiBMEC). We incorporated extracellular metabolite and isotope labeling data into an instationary metabolic flux analysis (INST-MFA) model to estimate intracellular metabolic fluxes. We found that NPC1 inhibition significantly increased glycolysis and pentose phosphate pathway flux while decreasing mitochondrial metabolism. These changes may have been driven by gene expression changes due to increased cholesterol biosynthesis, in addition to mitochondrial cholesterol accumulation. We corroborated these findings in primary BMEC, an alternative in vitro human brain endothelial model. Finally, we found that co-treatment with hydroxypropyl-β cyclodextrin (HPβCD) partially restored metabolic phenotype in U18666A-treated BMECs, suggesting that this drug may have therapeutic effects on the brain endothelium in NP-C. Together, our data highlight the importance of NPC1 in BMEC metabolism and implicate brain endothelial dysfunction in NP-C pathogenesis.http://www.sciencedirect.com/science/article/pii/S0969996124003711Niemann-pick disease type CNPC1MetabolismMetabolic flux analysisSystems biologyGlycolysis |
| spellingShingle | Bilal Moiz Matthew Walls Viviana Alpizar Vargas Anirudh Addepalli Callie Weber Andrew Li Ganesh Sriram Alisa Morss Clyne Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells Neurobiology of Disease Niemann-pick disease type C NPC1 Metabolism Metabolic flux analysis Systems biology Glycolysis |
| title | Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells |
| title_full | Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells |
| title_fullStr | Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells |
| title_full_unstemmed | Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells |
| title_short | Instationary metabolic flux analysis reveals that NPC1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells |
| title_sort | instationary metabolic flux analysis reveals that npc1 inhibition increases glycolysis and decreases mitochondrial metabolism in brain microvascular endothelial cells |
| topic | Niemann-pick disease type C NPC1 Metabolism Metabolic flux analysis Systems biology Glycolysis |
| url | http://www.sciencedirect.com/science/article/pii/S0969996124003711 |
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