Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis
Reticuloendothelial macrophages engulf ∼0.2 trillion senescent erythrocytes daily in a process called erythrophagocytosis (EP). This critical mechanism preserves systemic heme-iron homeostasis by regulating red blood cell (RBC) catabolism and iron recycling. Although extensive work has demonstrated...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-04-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphys.2020.00396/full |
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| author | Alexis Catala Alexis Catala Lyla A. Youssef Julie A. Reisz Monika Dzieciatkowska Nicholas E. Powers Carlo Marchetti Matthew Karafin James C. Zimring Krystalyn E. Hudson Kirk C. Hansen Steven L. Spitalnik Angelo D’Alessandro Angelo D’Alessandro |
| author_facet | Alexis Catala Alexis Catala Lyla A. Youssef Julie A. Reisz Monika Dzieciatkowska Nicholas E. Powers Carlo Marchetti Matthew Karafin James C. Zimring Krystalyn E. Hudson Kirk C. Hansen Steven L. Spitalnik Angelo D’Alessandro Angelo D’Alessandro |
| author_sort | Alexis Catala |
| collection | DOAJ |
| description | Reticuloendothelial macrophages engulf ∼0.2 trillion senescent erythrocytes daily in a process called erythrophagocytosis (EP). This critical mechanism preserves systemic heme-iron homeostasis by regulating red blood cell (RBC) catabolism and iron recycling. Although extensive work has demonstrated the various effects on macrophage metabolic reprogramming by stimulation with proinflammatory cytokines, little is known about the impact of EP on the macrophage metabolome and proteome. Thus, we performed mass spectrometry-based metabolomics and proteomics analyses of mouse bone marrow-derived macrophages (BMDMs) before and after EP of IgG-coated RBCs. Further, metabolomics was performed on BMDMs incubated with free IgG to ensure that changes to macrophage metabolism were due to opsonized RBCs and not to free IgG binding. Uniformly labeled tracing experiments were conducted on BMDMs in the presence and absence of IgG-coated RBCs to assess the flux of glucose through the pentose phosphate pathway (PPP). In this study, we demonstrate that EP significantly alters amino acid and fatty acid metabolism, the Krebs cycle, OXPHOS, and arachidonate-linoleate metabolism. Increases in levels of amino acids, lipids and oxylipins, heme products, and RBC-derived proteins are noted in BMDMs following EP. Tracing experiments with U-13C6 glucose indicated a slower flux through glycolysis and enhanced PPP activation. Notably, we show that it is fueled by glucose derived from the macrophages themselves or from the extracellular media prior to EP, but not from opsonized RBCs. The PPP-derived NADPH can then fuel the oxidative burst, leading to the generation of reactive oxygen species necessary to promote digestion of phagocytosed RBC proteins via radical attack. Results were confirmed by redox proteomics experiments, demonstrating the oxidation of Cys152 and Cys94 of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and hemoglobin-β, respectively. Significant increases in early Krebs cycle and C5-branched dibasic acid metabolites (α-ketoglutarate and 2-hydroxyglutarate, respectively) indicate that EP promotes the dysregulation of mitochondrial metabolism. Lastly, EP stimulated aminolevulinic acid (ALA) synthase and arginase activity as indicated by significant accumulations of ALA and ornithine after IgG-mediated RBC ingestion. Importantly, EP-mediated metabolic reprogramming of BMDMs does not occur following exposure to IgG alone. In conclusion, we show that EP reprograms macrophage metabolism and modifies macrophage polarization. |
| first_indexed | 2024-12-11T21:24:55Z |
| format | Article |
| id | doaj.art-8ab447cfcb524cb6a70f041c83dc87ca |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-11T21:24:55Z |
| publishDate | 2020-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj.art-8ab447cfcb524cb6a70f041c83dc87ca2022-12-22T00:50:22ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2020-04-011110.3389/fphys.2020.00396538149Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following ErythrophagocytosisAlexis Catala0Alexis Catala1Lyla A. Youssef2Julie A. Reisz3Monika Dzieciatkowska4Nicholas E. Powers5Carlo Marchetti6Matthew Karafin7James C. Zimring8Krystalyn E. Hudson9Kirk C. Hansen10Steven L. Spitalnik11Angelo D’Alessandro12Angelo D’Alessandro13Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesProgram in Structural Biology and Biochemistry, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesDepartment of Microbiology and Immunology, Columbia University, New York, NY, United StatesDepartment of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesDepartment of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesDepartment of Medicine – Division of Infectious Diseases, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesDepartment of Medicine – Division of Infectious Diseases, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesMedical Sciences Institute, Blood Center of Wisconsin (Versiti), Milwaukee, WI, United StatesDepartment of Pathology, University of Virginia, Charlottesville, VA, United StatesDepartment of Pathology and Cell Biology, Columbia University, New York, NY, United StatesDepartment of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesDepartment of Pathology and Cell Biology, Columbia University, New York, NY, United StatesDepartment of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesDepartment of Medicine – Division of Hematology, University of Colorado Denver – Anschutz Medical Campus, Aurora, CO, United StatesReticuloendothelial macrophages engulf ∼0.2 trillion senescent erythrocytes daily in a process called erythrophagocytosis (EP). This critical mechanism preserves systemic heme-iron homeostasis by regulating red blood cell (RBC) catabolism and iron recycling. Although extensive work has demonstrated the various effects on macrophage metabolic reprogramming by stimulation with proinflammatory cytokines, little is known about the impact of EP on the macrophage metabolome and proteome. Thus, we performed mass spectrometry-based metabolomics and proteomics analyses of mouse bone marrow-derived macrophages (BMDMs) before and after EP of IgG-coated RBCs. Further, metabolomics was performed on BMDMs incubated with free IgG to ensure that changes to macrophage metabolism were due to opsonized RBCs and not to free IgG binding. Uniformly labeled tracing experiments were conducted on BMDMs in the presence and absence of IgG-coated RBCs to assess the flux of glucose through the pentose phosphate pathway (PPP). In this study, we demonstrate that EP significantly alters amino acid and fatty acid metabolism, the Krebs cycle, OXPHOS, and arachidonate-linoleate metabolism. Increases in levels of amino acids, lipids and oxylipins, heme products, and RBC-derived proteins are noted in BMDMs following EP. Tracing experiments with U-13C6 glucose indicated a slower flux through glycolysis and enhanced PPP activation. Notably, we show that it is fueled by glucose derived from the macrophages themselves or from the extracellular media prior to EP, but not from opsonized RBCs. The PPP-derived NADPH can then fuel the oxidative burst, leading to the generation of reactive oxygen species necessary to promote digestion of phagocytosed RBC proteins via radical attack. Results were confirmed by redox proteomics experiments, demonstrating the oxidation of Cys152 and Cys94 of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and hemoglobin-β, respectively. Significant increases in early Krebs cycle and C5-branched dibasic acid metabolites (α-ketoglutarate and 2-hydroxyglutarate, respectively) indicate that EP promotes the dysregulation of mitochondrial metabolism. Lastly, EP stimulated aminolevulinic acid (ALA) synthase and arginase activity as indicated by significant accumulations of ALA and ornithine after IgG-mediated RBC ingestion. Importantly, EP-mediated metabolic reprogramming of BMDMs does not occur following exposure to IgG alone. In conclusion, we show that EP reprograms macrophage metabolism and modifies macrophage polarization.https://www.frontiersin.org/article/10.3389/fphys.2020.00396/fullmacrophage metabolismomics technologiesbloodOXPHOSmitochondrial dysregulationpentose phosphate pathway |
| spellingShingle | Alexis Catala Alexis Catala Lyla A. Youssef Julie A. Reisz Monika Dzieciatkowska Nicholas E. Powers Carlo Marchetti Matthew Karafin James C. Zimring Krystalyn E. Hudson Kirk C. Hansen Steven L. Spitalnik Angelo D’Alessandro Angelo D’Alessandro Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis Frontiers in Physiology macrophage metabolism omics technologies blood OXPHOS mitochondrial dysregulation pentose phosphate pathway |
| title | Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis |
| title_full | Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis |
| title_fullStr | Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis |
| title_full_unstemmed | Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis |
| title_short | Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis |
| title_sort | metabolic reprogramming of mouse bone marrow derived macrophages following erythrophagocytosis |
| topic | macrophage metabolism omics technologies blood OXPHOS mitochondrial dysregulation pentose phosphate pathway |
| url | https://www.frontiersin.org/article/10.3389/fphys.2020.00396/full |
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