Oxylipins_Perfusate

Oxilipidomic assessment of perfusate samples taken during 48-hour human kidney perfusion belonging to the project "Normothermic human kidney preservation drives iron accumulation and ferroptosis"<b>Abstract</b><br>Ex vivo normothermic machine perfusion has been proposed to protect deceased donor kidneys. However, its benefits remain ambiguous. We postulate that the use of red blood cells (RBCs) and associated secondary hemolysis may in fact cause renal injury, offsetting potential advantages. During 48-hour normothermic perfusion of seven human donor kidneys, we observed progressive hemolysis, leading to iron accumulation in perfusate, tissue, and urine. Untargeted lipidomic profiling revealed significant increases in oxidized phospholipid species in perfused kidneys, pointing towards iron-dependent cell death known as ferroptosis. Next, in twelve additional perfusions, we assessed strategies to mitigate hemolysis-driven injury. Dialysis-based free hemoglobin removal reduced lipid peroxidation, but a ferroptosis gene signature persisted. In contrast, cell-free perfusion at subnormothermia negated iron accumulation, the ferroptosis gene signature, phospholipid peroxidation, and acute kidney injury. Our findings highlight the pathological role of hemolysis and iron on the kidney, urging restraint in the clinical application of RBC-based kidney perfusion.

本数据集源自"常温人体肾脏保存可诱导铁积累与铁死亡（Normothermic human kidney preservation drives iron accumulation and ferroptosis）"项目，包含7例人体供肾在48小时常温机器灌注（normothermic machine perfusion）过程中采集的灌注液样本的氧化脂组学（oxilipidomic）评估数据。<br><b>摘要</b><br>离体常温机器灌注曾被提议用于保护遗体供肾，但其临床获益仍不明确。我们提出假设：灌注液中使用红细胞（red blood cells, RBCs）及其继发的溶血（hemolysis）反应反而可能造成肾损伤，抵消其潜在优势。在7例人体供肾的48小时常温灌注过程中，我们观察到进行性溶血反应，导致灌注液、肾组织及尿液中出现铁积累。非靶向脂质组学分析（untargeted lipidomic profiling）显示，灌注后的肾脏中氧化磷脂类物质显著升高，这指向一种铁依赖的细胞死亡形式——铁死亡（ferroptosis）。随后，在另外12例灌注实验中，我们评估了缓解溶血介导损伤的策略：基于透析的游离血红蛋白清除手段可减轻脂质过氧化（lipid peroxidation），但铁死亡基因特征仍持续存在。与之相反，亚常温（subnormothermia）无细胞灌注（cell-free perfusion）可完全消除铁积累、铁死亡基因特征、磷脂过氧化及急性肾损伤（acute kidney injury）。我们的研究结果揭示了溶血与铁对肾脏的病理作用，呼吁临床中谨慎使用基于红细胞的肾脏灌注方案。