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
View: Text | PDF
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

×

Figure 6

Storage lesions occurring during aging in vitro are concentrated in the long-stored CFSEhi RBC subset; these are preferentially cleared during ex vivo human spleen perfusion.

Options: View larger image (or click on image) Download as PowerPoint
Storage lesions occurring during aging in vitro are concentrated in the ...
(A) Retention rate by microsphiltration, (B) dynamic RBC adhesion to endothelial cells, (C) RBC surface PS exposure quantified by lactadherin staining, (D) osmotic fragility determined by measuring the NaCl concentration required to induce 50% hemolysis, (E) intracellular ATP levels, normalized to hemoglobin content, measured with CFSE-stained unsorted long-stored RBCs (LS-unsorted) RBCs and on flow-sorted LS-CFSElo and LS-CFSEhi RBCs. In A–E, data are represented as individual points with mean ± SEM of ≥6 RBC concentrates). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by Friedman’s 1-way ANOVA followed by Dunn’s multiple comparison test. In C, CTV-stained RBCs were analyzed to allow lactadherin-FITC staining. (F) Kinetics (mean ± SEM of 4 independent perfusions) of the normalized circulating concentrations of stained short-stored (SS), long-stored (LS), LS-CFSElo, and LS-CFSEhi RBC subsets during ex vivo perfusion of human spleen. In F, a 2-way ANOVA followed by Šidák’s multiple comparison test was performed. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and ####P < 0.0001 by a 2-way ANOVA followed by Šidák’s multiple comparison test. Different symbols are used to show statistical significance: asterisks are used for comparisons of SS vs. LS and pound signs are used for comparisons of LS-CFSElo vs. LS-CFSEhi. No statistically significant differences were seen when comparing SS-CFSElo vs. LS-CFSElo.