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Unlike nucleated cells where multiple hallmarks characterize aging, proteostasis is expected to play a main role in anucleate RBCs, including proteasomal protein degradation. Although proteasomal activity declines during aging in vitro, its role in generating downstream alterations and post-transfusion clearance remains unclear. We hypothesized that proteasome inhibition accelerates RBC aging in vitro, particularly following re-exposure to physiological conditions. We evaluated the impact of proteasome inhibition (i.e., epoxomicin) on RBC quality during storage and physiological restoration in vitro. Additionally, young and old RBC subpopulations were compared. Proteasome inhibition during storage depleted ATP and altered RBC morphology without immediate oxidative damage. Physiological restoration of proteasome-inhibited RBCs caused accelerated ATP depletion, massive protein aggregation, reduced deformability, hemolysis, and phosphatidylserine exposure, particularly in long-stored RBCs. Strikingly, RBCs aged in vivo also exhibited low proteasomal activity and behaved similarly to stored RBCs following physiological restoration. In conclusion, proteasomal dysfunction is a key hallmark of RBC aging and senescence, driving molecular and cellular modifications that mark RBCs for clearance in vivo. Therefore, enhancing proteasomal function could improve RBC storage quality and transfusion efficacy.<\/jats:p>","DOI":"10.3390\/antiox15091146","type":"journal-article","created":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T16:43:16Z","timestamp":1788972196000},"page":"1146","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Proteasome Safeguards Red Blood Cell Integrity During Storage and After Transfusion"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5520-7346","authenticated-orcid":false,"given":"Sandy","family":"Peltier","sequence":"first","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, INSERM, EFS, BIGR U1134, Team BioTiGR, 75015 Paris, France"},{"name":"Universit\u00e9 Paris Cit\u00e9, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, INSERM, 75015 Paris, France"},{"name":"Initiatives IdEx Globule Rouge d\u2019Excellence (InIdex GR-Ex), Universit\u00e9 Paris Cit\u00e9, 75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"given":"Th\u00e9o","family":"Michel","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interr\u00e9gionale CRS, 1066 Epalinges, Switzerland"},{"name":"Center for Research and Innovation in Clinical Pharmaceutical Sciences, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Fanny","family":"Mialane","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, INSERM, EFS, BIGR U1134, Team BioTiGR, 75015 Paris, France"},{"name":"Initiatives IdEx Globule Rouge d\u2019Excellence (InIdex GR-Ex), Universit\u00e9 Paris Cit\u00e9, 75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4094-022X","authenticated-orcid":false,"given":"Micka\u00ebl","family":"Marin","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, INSERM, EFS, BIGR U1134, Team BioTiGR, 75015 Paris, France"},{"name":"Initiatives IdEx Globule Rouge d\u2019Excellence (InIdex GR-Ex), Universit\u00e9 Paris Cit\u00e9, 75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Micha\u00ebl","family":"Dussiot","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, INSERM, 75015 Paris, France"},{"name":"Initiatives IdEx Globule Rouge d\u2019Excellence (InIdex GR-Ex), Universit\u00e9 Paris Cit\u00e9, 75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"C\u00e9line","family":"Rodriguez","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interr\u00e9gionale CRS, 1066 Epalinges, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Monika","family":"Dzieciatkowska","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, University of Colorado Denver, Anschutz Medical Campus, Aurora, CO 80045, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8969-8166","authenticated-orcid":false,"given":"Marie","family":"Tamagne","sequence":"additional","affiliation":[{"name":"EFS Ile de France\u2014Institut Mondor, INSERM, Universit\u00e9 Paris-Est Cr\u00e9teil Val de Marne, Eq. 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