Proteostasis and metabolic dysfunction characterize a subset of storage-induced senescent erythrocytes targeted for posttransfusion clearance
Sandy Peltier, Mickaël Marin, Monika Dzieciatkowska, Michaël Dussiot, Micaela Kalani Roy, Johanna Bruce, Louise Leblanc, Youcef Hadjou, Sonia Georgeault, Aurélie Fricot, Camille Roussel, Daniel Stephenson, Madeleine Casimir, Abdoulaye Sissoko, François Paye, Safi Dokmak, Papa Alioune Ndour, Philippe Roingeard, Emilie-Fleur Gautier, Steven L. Spitalnik, Olivier Hermine, Pierre A. Buffet, Angelo D’Alessandro, Pascal Amireault
Sandy Peltier, Mickaël Marin, Monika Dzieciatkowska, Michaël Dussiot, Micaela Kalani Roy, Johanna Bruce, Louise Leblanc, Youcef Hadjou, Sonia Georgeault, Aurélie Fricot, Camille Roussel, Daniel Stephenson, Madeleine Casimir, Abdoulaye Sissoko, François Paye, Safi Dokmak, Papa Alioune Ndour, Philippe Roingeard, Emilie-Fleur Gautier, Steven L. Spitalnik, Olivier Hermine, Pierre A. Buffet, Angelo D’Alessandro, Pascal Amireault
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Research Article Cell biology Hematology

Proteostasis and metabolic dysfunction characterize a subset of storage-induced senescent erythrocytes targeted for posttransfusion clearance

Abstract

Although refrigerated storage slows the metabolism of volunteer donor RBCs, which is essential in transfusion medicine, cellular aging still occurs throughout this in vitro process. Storage-induced microerythrocytes (SMEs) are morphologically altered senescent RBCs that accumulate during storage and are cleared from circulation following transfusion. However, the molecular and cellular alterations that trigger clearance of this RBC subset remain to be identified. Using a staining protocol that sorts long-stored SMEs (i.e., CFSEhi) and morphologically normal RBCs (CFSElo), these in vitro aged cells were characterized. Metabolomics analysis identified depletion of energy, lipid-repair, and antioxidant metabolites in CFSEhi RBCs. By redox proteomics, irreversible protein oxidation primarily affected CFSEhi RBCs. By proteomics, 96 proteins, mostly in the proteostasis family, had relocated to CFSEhi RBC membranes. CFSEhi RBCs exhibited decreased proteasome activity and deformability; increased phosphatidylserine exposure, osmotic fragility, and endothelial cell adherence; and were cleared from the circulation during human spleen perfusion ex vivo. Conversely, molecular, cellular, and circulatory properties of long-stored CFSElo RBCs resembled those of short-stored RBCs. CFSEhi RBCs are morphologically and metabolically altered, have irreversibly oxidized and membrane-relocated proteins, and exhibit decreased proteasome activity. In vitro aging during storage selectively alters metabolism and proteostasis in these storage-induced senescent RBCs targeted for clearance.

Authors

Sandy Peltier, Mickaël Marin, Monika Dzieciatkowska, Michaël Dussiot, Micaela Kalani Roy, Johanna Bruce, Louise Leblanc, Youcef Hadjou, Sonia Georgeault, Aurélie Fricot, Camille Roussel, Daniel Stephenson, Madeleine Casimir, Abdoulaye Sissoko, François Paye, Safi Dokmak, Papa Alioune Ndour, Philippe Roingeard, Emilie-Fleur Gautier, Steven L. Spitalnik, Olivier Hermine, Pierre A. Buffet, Angelo D’Alessandro, Pascal Amireault

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Figure 4

Proteomics identifies subset-specific membrane relocation of specific proteins in long-stored CFSEhi RBCs, notably affecting proteins in the proteostasis family.

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Proteomics identifies subset-specific membrane relocation of specific pr...
(A) PCA of proteomics data obtained from membrane preparations isolated from flow-sorted short-stored CFSElo (SS-CFSElo, in red), long-stored CFSElo (LS-CFSElo, light green), and long-stored CFSEhi (LS-CFSEhi, dark green) RBCs. (B) Hierarchical clustering analysis of the 96 proteins that significantly differ between the 3 groups following Pearson’s clusterization and a FDR correction without data imputation (proteins detected in at least 70% of samples were considered). A z-score scale calculated from copy number/cell shows each protein level, while lack of detection is represented by a gray area. (C) Interaction network analysis for proteins increased (full filled circle) and decreased (white, transparent, inner circle) in membrane preparations of long-stored CFSEhi RBCs, as compared with long-stored CFSElo RBCs. Gray lines represent physical interactions between proteins. This network was realized using Cytoscape StringApp 3.9. Colored dotted lines represent the main protein families identified (relative proportion within the 96 significant proteins), and colored circles represent functional groups within a family.