Effect of blood donor sex and testosterone on predisposition to hemolysis in stored red blood cells

项目摘要 献血者是一个遗传多样性的群体，有许多生物变量，包括性别、种族和 年龄可能会影响红细胞(RBC)的储存和输血结果。我们已经演示了性爱 红细胞溶血易感性的二分性，人或鼠的雄性红细胞表现出增强的 对冷藏、渗透性休克和氧化应激的敏感性。我们研究的一个重要发现是 小鼠睾丸切除或睾酮替代疗法(TRT)显著调节红细胞易感性 储存中和输血后的溶血。这一发现提醒我们注意与 献血者中的TRT。睾丸激素疗法最近被认为是最被滥用的疗法之一。 美国的医疗实践，以及通过TRT输注红细胞缺乏明确的指南和风险评估 患者认为这些捐赠者有资格捐献，但疑似红细胞增多症的病例除外。我们的预赛 对25个小鼠品系的全基因组关联研究表明，性别差异对易感性的影响 溶血涉及p38MAPK周围的基因网络，这可能会在 应激，并可被睾丸素激活。这些观察结果告诉我们最重要的假设是 睾酮调节红系细胞分化和生物学导致红细胞性别差异 包括冷藏在内的溶血应激条件下的特性和激酶活性。在目前的提案中，我们 使用创新的人类和小鼠研究来评估供者性别和睾酮对输血的影响 结果并绘制出调节RBC结构、功能的下游p38 MAPK信号通路 以及储存和输血过程中的完整性。在目标1中，我们将定义献血者TRT对红细胞的影响 使用人到小鼠输血模型的储存和输血后恢复。在目标2中，我们将确定 睾酮与p38MAPK的分子相互作用及p38MAPK信号中枢的确定 会影响红细胞的功能和在储存中的存活率。睾酮对p38MAPK生物学的影响(目标2A) 将使用人类红系细胞系在不同水平的红系细胞分化中进行定义；成熟时 TRT患者在睾酮治疗前和治疗后120天的红细胞；以及小鼠的红细胞 表达减弱的p38MAPK活性(B6.Cg-Mapk14tm1.1Dvb/J)。在所有实验中，我们将监测 P38MAPK下游包括MK2、HSP27和阴离子交换蛋白R-1的蛋白表达变化。下一首, 我们将使用小鼠和捐赠者(NHLBI RBC-OMICS)基因组范围关联(GWA)的性别数据库 溶血差异促进发现与p38 MAPK相关并受其调控的信号枢纽 性。这个项目的成果可能会通过评估风险来推动输血医学领域的发展。 与输注TRT捐献者的血液有关；为 调节红细胞储存稳定性的性别差异；并确定可以作为靶点的信号枢纽 在治疗方面，以减少输血实践中的溶血风险。

Project Summary Blood donors are a genetically diverse population with numerous biological variables including sex, race, and age that may affect red blood cell (RBC) storage and transfusion outcomes. We have demonstrated a sex dichotomy in RBC predisposition to hemolysis, for which male RBCs from humans or mice exhibit enhanced susceptibility to cold storage, osmotic shock, and oxidative stress. A key discovery of our studies is that orchiectomy or testosterone replacement therapy (TRT) in mice significantly modulates RBC predisposition to hemolysis in storage and after transfusion. This finding has alerted us to the potential risks associated with TRT in blood donors. Testosterone therapies have been recently identified as of one of the most overused medical practices in the US, and the lack of clear guidelines and risk assessment of RBC transfusion from TRT patients deem such donors eligible for donation, except for cases of suspected polycythemia. Our preliminary genome-wide association studies in 25 mouse strains suggested that sex differences in predisposition to hemolysis involve gene networks surrounding p38 MAPK, which may intensify hemolytic response during stress, and can be activated by testosterone. These observations inform our overarching hypothesis that testosterone modulates erythroid cell differentiation and biology leading to sex differences in RBC characteristics and kinase activity under hemolytic stress including cold storage. In the current proposal, we use innovative human and mouse studies to evaluate the impact of donor sex and testosterone on transfusion outcomes and to map out down-stream p38 MAPK signaling pathways that modulate RBC structure, function and integrity during storage and transfusion. In Aim 1, we will define the impact of TRT in blood donors on RBC storage and post transfusion recovery using a human to mouse transfusion model. In Aim 2, we will determine the molecular interactions between testosterone and p38 MAPK, and identify the p38 MAPK signaling hubs that impact RBC function and survival in storage. The impact of testosterone on p38 MAPK biology (Aim 2A) will be defined at different levels of erythroid cell differentiation using human erythroid cell lines; in mature RBCs from TRT patients before and 120 days after testosterone treatments; and in RBCs from mice expressing attenuated p38 MAPK activity (B6.Cg-Mapk14tm1.1Dvb/J). In all experiments, we will monitor for changes in protein expression downstream of p38 MAPK including MK2, HSP 27 and anion exchangr-1. Next, we will use mouse and donor (NHLBI RBC-Omics) genome-wide association (GWA) databases of sex differences in hemolysis to enhance discovery of signaling hubs associated with p38 MAPK and regulated by sex. Outcomes from this project are likely to advance the field of transfusion medicine by assessing the risks associated with transfusion of blood from TRT donors; providing new insights into the mechanisms that mediate sex differences in RBC storage stability; and identifying signaling hubs that can be targeted therapeutically to reduce the risk of hemolysis in transfusion practices.