Nemkov Travis, Yoshida Tatsuro, Nikulina Maria, D'Alessandro Angelo
Omix Technologies Inc., Denver, CO, United States.
Department of Biochemistry and Molecular Genetics, University of Colorado Denver - Anschutz Medical Campus, Aurora, CO, United States.
Front Physiol. 2022 Mar 14;13:833242. doi: 10.3389/fphys.2022.833242. eCollection 2022.
Red blood cell transfusion is a life-saving intervention, and storage is a logistic necessity to make ~110 million units available for transfusion every year worldwide. However, storage in the blood bank is associated with a progressive metabolic decline, which correlates with the accumulation of morphological lesions, increased intra- and extra-vascular hemolysis upon transfusion, and altered oxygen binding/off-loading kinetics. Prior to storage, red blood cells are suspended in nutrient formulations known as additive solutions to prolong cellular viability. Despite a thorough expansion of knowledge regarding red blood cell biology over the past few decades, only a single new additive solution has been approved by the Food and Drug Administration this century, owing in part to the limited capacity for development of novel formulations. As a proof of principle, we leveraged a novel high-throughput metabolomics technology as a platform for rapid data-driven development and screening of novel additive solutions for blood storage under both normoxic and hypoxic conditions. To this end, we obtained leukocyte-filtered red blood cells (RBCs) and stored them under normoxic or hypoxic conditions in 96 well plates (containing polyvinylchloride plasticized with diethylhexylphthalate to concentrations comparable to full size storage units) in the presence of an additive solution supplemented with six different compounds. To inform this data-driven strategy, we relied on previously identified metabolic markers of the RBC storage lesion that associates with measures of hemolysis and post-transfusion recovery, which are the FDA gold standards to predict stored blood quality, as well as and metabolic predictors of oxygen binding/off-loading parameters. Direct quantitation of these predictors of RBC storage quality were used here-along with detailed pathway analysis of central energy and redox metabolism-as a decision-making tool to screen novel additive formulations in a multiplexed fashion. Candidate supplements are shown here that boost-specific pathways. These metabolic effects are only in part dependent on the SO storage conditions. Through this platform, we anticipate testing thousands of novel additives and combinations thereof in the upcoming months.
红细胞输血是一种挽救生命的干预措施，而储存是一项后勤必需工作，以便每年在全球范围内提供约1.1亿单位的可用于输血的红细胞。然而，血库中的储存与红细胞代谢的逐渐衰退有关，这与形态学损伤的积累、输血时血管内和血管外溶血的增加以及氧结合/释放动力学的改变相关。在储存之前，红细胞悬浮于称为添加剂溶液的营养配方中以延长细胞活力。尽管在过去几十年中对红细胞生物学的认识有了全面扩展，但本世纪美国食品药品监督管理局仅批准了一种新的添加剂溶液，部分原因是新型配方的开发能力有限。作为原理验证，我们利用一种新型高通量代谢组学技术作为平台，用于在常氧和低氧条件下快速进行数据驱动的新型血液储存添加剂溶液的开发和筛选。为此，我们获取了白细胞滤除的红细胞，并将其在常氧或低氧条件下保存在96孔板中（孔板含有邻苯二甲酸二己酯增塑的聚氯乙烯，其浓度与全尺寸储存单元相当），孔板中含有添加了六种不同化合物的添加剂溶液。为了为这种数据驱动策略提供信息，我们依赖于先前确定的与溶血和输血后恢复指标相关的红细胞储存损伤的代谢标志物，这些指标是美国食品药品监督管理局预测储存血液质量的金标准，以及氧结合/释放参数的代谢预测指标。此处使用这些红细胞储存质量预测指标的直接定量分析以及中心能量和氧化还原代谢的详细途径分析，作为以多重方式筛选新型添加剂配方的决策工具。此处展示了能促进特定途径的候选补充剂。这些代谢效应仅部分依赖于储存条件。通过这个平台，我们预计在未来几个月内测试数千种新型添加剂及其组合。
