Peltier Sandy, Marin Mickaël, Dzieciatkowska Monika, Dussiot Michaël, Roy Micaela Kalani, Bruce Johanna, Leblanc Louise, Hadjou Youcef, Georgeault Sonia, Fricot Aurélie, Roussel Camille, Stephenson Daniel, Casimir Madeleine, Sissoko Abdoulaye, Paye François, Dokmak Safi, Ndour Papa Alioune, Roingeard Philippe, Gautier Emilie-Fleur, Spitalnik Steven L, Hermine Olivier, Buffet Pierre A, D'Alessandro Angelo, Amireault Pascal
Université Paris Cité, INSERM, BIGR, Paris, France.
Université Paris Cité, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, INSERM, Paris, France.
J Clin Invest. 2025 Mar 11;135(9). doi: 10.1172/JCI183099. eCollection 2025 May 1.
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.
尽管冷藏储存会减缓自愿献血者红细胞的新陈代谢（这在输血医学中至关重要），但在这个体外过程中细胞衰老仍会发生。储存诱导的微红细胞（SMEs）是形态改变的衰老红细胞，在储存过程中积累，并在输血后从循环中清除。然而，触发清除这一红细胞亚群的分子和细胞改变仍有待确定。使用一种染色方案对长期储存的SMEs（即CFSEhi）和形态正常的红细胞（CFSElo）进行分类，对这些体外老化细胞进行了表征。代谢组学分析确定了CFSEhi红细胞中能量、脂质修复和抗氧化代谢物的消耗。通过氧化还原蛋白质组学，不可逆的蛋白质氧化主要影响CFSEhi红细胞。通过蛋白质组学，96种蛋白质（大多属于蛋白质稳态家族）重新定位到CFSEhi红细胞膜上。CFSEhi红细胞表现出蛋白酶体活性和变形性降低；磷脂酰丝氨酸暴露、渗透脆性和内皮细胞黏附增加；并在体外人脾灌注过程中从循环中清除。相反，长期储存的CFSElo红细胞的分子、细胞和循环特性与短期储存的红细胞相似。CFSEhi红细胞在形态和代谢上发生改变，具有不可逆氧化和膜重新定位的蛋白质，并表现出蛋白酶体活性降低。储存期间的体外老化选择性地改变了这些靶向清除的储存诱导衰老红细胞的代谢和蛋白质稳态。
