PXD017788

PXD017788 is an original dataset announced via ProteomeXchange.

Title	Mouse BMDMs and Erythrophagocytosis LCMSMS
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 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.
HostingRepository	PRIDE
AnnounceDate	2024-10-07
AnnouncementXML	Submission_2024-10-07_10:37:10.215.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD017788
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Alexis Catala
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	amidated residue; dehydrated residue; Cys->Dha; Oxidation; Dioxidation; Acetyl; Carbamidomethyl; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue; Gln->pyro-Glu; deamidated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2020-02-28 08:05:10	ID requested
⏵ 1	2024-10-07 10:37:11	announced

10.6019/PXD017788;

10.3389/fphys.2020.00396;

Catala A, Youssef LA, Reisz JA, Dzieciatkowska M, Powers NE, Marchetti C, Karafin M, Zimring JC, Hudson KE, Hansen KC, Spitalnik SL, D'Alessandro A, Metabolic Reprogramming of Mouse Bone Marrow Derived Macrophages Following Erythrophagocytosis. Front Physiol, 11():396(2020) [pubmed]

submitter keyword: Mouse, RBCs, BMDMs, LCMSMS

Angelo D'Alessandro
contact affiliation	University of Colorado Denver – Anschutz Medical Campus, Department of Biochemistry and Molecular Genetics, Aurora, CO, USA
contact email	angelo.dalessandro@cuanschutz.edu
lab head
Alexis Catala
contact affiliation	University of Colorado Denver-Anschutz Medical Campus
contact email	alexis.catala@cuanschutz.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/05/PXD017788

PRIDE project URI

• PRIDE
  1. PXD017788
     1. Label: PRIDE project
     2. Name: Mouse BMDMs and Erythrophagocytosis LCMSMS