Project 1: Identification and validation of biomarkers for RBC metabolic aging

输注红细胞（RBC）是美国医院最常见的治疗程序， 有效预防贫血患者的发病和死亡。然而，储存的红细胞单位 逐渐发展称为“RBC储存病变”的功能缺陷，可能影响临床 结果。供体特异性因素也可能决定储存红细胞的输血效果。因为 由于储存时间和/或供体因素，输注有缺陷的红细胞单位可能会产生不利影响， 每年影响成千上万的患者，应该制定方法来筛选这些单位， 血液供应代谢组学是鉴定与代谢相关的生物标志物的有力方法。 以及储存的RBC中的功能变化。为了支持我们的目标，以确定代谢生物标志物， 通过供体检测，我们现在已经证明，来自> 90%供体的AS 1-RBC显示出典型的变化， 在42天的储存过程中，数百种代谢物（“典型代谢型”），而其余的代谢物 供体具有明显不同的代谢型。有趣的是，没有单一的非典型代谢型：所有 迄今为止，我们所测试的具有非典型代谢型的供体彼此显著不同。因此，在本发明中， 我们认为，这些不同的代谢型区分了一小部分储存的红细胞， 代谢/功能异常值，并且在其RBC中鉴定的代谢改变可用于 开发生物标志物来检测输血后具有不同生存或功效特征的红细胞。在 在拟议的研究中，我们将利用来自已知储存不良供体的红细胞来识别代谢 这些生物标志物与储存期间体外功能障碍的几种测量相关。这些 然后，生物标志物将用于筛选随机捐赠者，其中一些人也将接受体内检测。 输血后的RBC存活率，以测试候选生物标志物（最初鉴定为 从已知的存储差的供体）可以依次识别随机选择的具有存储差的供体 特色这项工作的成功完成将导致一系列红细胞的鉴定 预测血液储存期间RBC功能改变的生物标志物，可进一步验证 临床使用，并可作为一个起点，剖析机制的基础储存不良， 捐献的RBC的特征。

Transfusion of red blood cells (RBCs), the most common therapeutic procedure in US hospitals, is effective at preventing morbidity and mortality in anemic patients. However, stored RBC units progressively develop functional defects known as "RBC storage lesions" that may affect clinical outcomes. Donor-specific factors may also determine the transfusion efficacy of stored RBCs. Because transfusion of RBC units that are defective, due to storage time and/or donor factors, may adversely affect thousands of patients annually, approaches should be developed to screen these units from the blood supply. Metabolomics is a powerful approach to identify biomarkers that correlate with metabolic and functional changes in stored RBCs. In support of our objective to identify metabolic biomarkers for donor testing, we have now demonstrated that AS1-RBCs from > 90% of donors show typical changes in hundreds of metabolites during 42 day storage (the "typical metabotype), whereas the remaining donors have markedly divergent metabotypes. Intriguingly, there is not single atypical metabotype: all donors we have tested to date with atypical metabotypes differed significantly from one another. Thus, we propose that these divergent metabotypes differentiate a small percentage of stored RBCs as metabolic/functional outliers, and that metabolic alterations identified in their RBCs can be used to develop biomarkers to detect RBCs with different survival or efficacy characteristics after transfusion. In the proposed studies, we will utilize RBCs from known-poor storing donors to identify metabolic biomarkers that correlate with several measures of in vitro dysfunction during storage. These biomarkers will then be used to screen random donors, some of whom will also be tested for in vivo RBC survival after transfusion to test whether the candidate biomarkers (which were initially identified from known poor-storing donors) can in turn identify randomly selected donors with poor-storage characteristics. Successful completion of this work will result in identification of a series of RBC biomarkers that predict altered RBC function during blood storage, that can be further validated for clinical use, and that can be used as a starting point to dissect mechanisms underlying poor-storage characteristics of donated RBCs.