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 7

A proposed model to illustrate the main alterations affecting each RBC subset during storage.

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A proposed model to illustrate the main alterations affecting each RBC s...
The main results of the comparative molecular and cellular characterizations (black boxes) of long-stored CFSElo (light green, discocytes) and long-stored CFSEhi (dark green, echinocytes III, spheroechinocytes, and spherocytes) RBCs. In long-stored CFSElo RBCs (left), functional energy and redox metabolism sustains effective lipid-repair and proteostasis functions, limiting protein aggregation, membrane relocation, and vesiculation. Functional molecular properties contribute to maintaining normal cellular properties (e.g., morphology, deformability, endothelial cell adhesion) and, thus, their ability to circulate. In long-stored CFSEhi RBCs (right), decreased energy metabolism (e.g., ATP, PP, GAPD) is unable to fuel the redox system effectively, leading to decreased GSH and increased GSSG levels, culminating in dysfunction in lipid repair and proteostasis. The accumulation of irreversibly oxidized proteins could also lead to decreased proteasome function. The combined effect of oxidized protein accumulation and decreased proteasome degradation could fuel a negative feedback loop that further inhibits the redox system, thereby favoring production of toxic protein aggregates, protein relocation to the RBC membrane, and, ultimately, vesiculation. Vesiculation alters morphology, leading to decreased deformability, increased endothelial cell adhesion, and splenic retention. Mild (light blue boxes) and marked alterations are shown (pink boxes); black and red lines represent normal and negative interactions, respectively. PP, pentose phosphate, GADP, DL-GAPDH; GSH, reduced glutathione; GSSG, oxidized glutathione; Acyl-C, acyl-carnitines; Reversible/Irreversible Ox, reversible/irreversible oxidation.