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

献血者性别和睾酮对储存红细胞溶血倾向的影响

• 批准号: 10322992
• 负责人: Tamir Kanias
• 金额: $ 38.52万
• 依托单位: VITALANT
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2023-09-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322992
• 关键词: Affect; Age; American; Androgens; Anions; Apoptosis; Attenuated; Binding; Biological; Biological Factors; Biology; Blood Banks; Blood Circulation; Blood Donations; Blood Transfusion; Blood donor; Candidate Disease Gene; Cell Differentiation process; Cell Line; Cell Survival; Cell physiology; Cellular Structures; Cellular biology; Characteristics; Code; Cryopreservation; Databases; Disease; Erythrocyte Transfusion; Erythrocytes; Erythroid Cells; Erythropoiesis; Exhibits; Gene Expression; Genes; Genetic; Genetic Databases; Genetic Predisposition to Disease; Genomics; Goals; Guidelines; Hemolysis; Human; Human Genome; Knock-out; MAP Kinase Gene; Maps; Mediating; Mediator of activation protein; Medical; Membrane; Modeling; Molecular; Monitor; Mouse Strains; Mus; National Heart, Lung, and Blood Institute; Orchiectomy; Osmotic Shocks; Outcome; Ovariectomy; Oxidative Stress; Patients; Pharmaceutical Preparations; Phosphotransferases; Polycythemia; Population Heterogeneity; Population Study; Predisposition; Race; Recovery; Risk; Risk Assessment; Safety; Sex Differences; Signal Pathway; Signal Transduction; Stress; Technology; Testing; Testosterone; Therapeutic; Transfusion; clinically significant; erythroid differentiation; ethnic diversity; experimental study; gene network; genome wide association study; improved; individual patient; innovation; insight; male; men; mouse genome; p38 MAPK Signaling Pathway; p38 Mitogen Activated Protein Kinase; protein expression; resilience; response; sex; targeted treatment; testosterone replacement therapy; transfusion medicine

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.

项目摘要 献血者是一个遗传多样性的人群，具有许多生物学变量，包括性别，种族， 可能影响红细胞（RBC）储存和输血结果的年龄。我们展示了一种性别 红细胞溶血倾向的二分法，其中来自人或小鼠的雄性红细胞表现出增强的溶血倾向。 对冷藏、渗透压休克和氧化应激的易感性。我们研究的一个关键发现是， 睾丸切除术或睾酮替代疗法（TRT）可显著调节小鼠红细胞倾向， 储存和输血后溶血。这一发现提醒我们， 献血者中的TRT。最近，替吉奥疗法被认为是最被滥用的疗法之一。 美国的医疗实践，以及缺乏明确的指导方针和TRT红细胞输注的风险评估 患者认为这些捐献者符合捐献条件，但疑似红细胞增多症的情况除外。我们的初步 对25种小鼠品系的全基因组关联研究表明，易感性存在性别差异 溶血涉及p38 MAPK周围基因网络，其可在溶血过程中增强溶血反应， 压力，并可由睾酮激活。这些观察为我们的总体假设提供了依据， 睾酮调节红系细胞分化和生物学，导致RBC中的性别差异 在溶血胁迫（包括冷藏）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体性别和睾酮对输血的影响 结果，并绘制出下游p38 MAPK信号通路，调节RBC结构，功能， 以及在储存和输注过程中的完整性。在目标1中，我们将定义TRT在献血者中对RBC的影响。 储存和使用人-小鼠输注模型的输注后回收。在目标2中，我们将确定 睾酮和p38 MAPK之间的分子相互作用，并确定p38 MAPK信号传导枢纽 影响红细胞功能和储存存活率。睾酮对p38 MAPK生物学的影响（Aim 2A） 将使用人红系细胞系在不同水平的红系细胞分化下定义;在成熟红系细胞中， 睾酮治疗前和治疗后120天TRT患者的RBC;以及小鼠的RBC 表达减弱的p38 MAPK活性（B6. Cg-Mapk 14 tm 1.1Dvb/J）。在所有实验中，我们将监测 p38 MAPK下游蛋白表达的变化，包括MK2、HSP 27和阴离子交换剂r-1。接下来， 我们将使用小鼠和供体（NHLBI RBC组学）全基因组关联（GWA）性别数据库， 溶血的差异，以增强发现与p38 MAPK相关的信号中枢，并受 性该项目的结果可能会通过评估风险来推动输血医学领域的发展 与TRT献血者输血相关;提供了对TRT机制的新见解， 介导RBC储存稳定性的性别差异;并确定可以靶向的信号中枢 在治疗上减少输血实践中溶血的风险。

项目成果

Sex-specific genetic modifiers identified susceptibility of cold stored red blood cells to osmotic hemolysis.

• DOI: 10.1186/s12864-022-08461-4
• 发表时间: 2022-03-23
• 期刊: BMC genomics
• 影响因子: 4.4
• 作者: Fang F;Hazegh K;Mast AE;Triulzi DJ;Spencer BR;Gladwin MT;Busch MP;Kanias T;Page GP
• 通讯作者: Page GP

Relationships between endogenous and exogenous testosterone and cardiovascular disease in men.

男性内源性和外源睾丸激素与心血管疾病之间的关系。

• DOI: 10.1007/s11154-022-09752-7
• 发表时间: 2022-12
• 期刊: REVIEWS IN ENDOCRINE & METABOLIC DISORDERS
• 影响因子: 8.2
• 作者: Thirumalai, Arthi;Anawalt, Bradley D.
• 通讯作者: Anawalt, Bradley D.

Toxic masculinity in red blood cell units? Testosterone therapy in blood donors revisited.

• DOI: 10.1111/trf.16658
• 发表时间: 2021-11
• 期刊: Transfusion
• 影响因子: 2.9
• 作者: Hazegh K;Anawalt BD;Dumont LJ;Kanias T
• 通讯作者: Kanias T

Erythrocyte mitogen-activated protein kinases mediate hemolytic events under osmotic and oxidative stress and in hemolytic diseases.

• DOI: 10.1016/j.cellsig.2022.110450
• 发表时间: 2022-11
• 期刊: CELLULAR SIGNALLING
• 影响因子: 4.8
• 作者: Hazegh, Kelsey;Fang, Fang;Kelly, Kathleen;Sinchar, Derek;Wang, Ling;Zuchelkowski, Benjamin E;Ufelle, Alexander C;Esparza, Orlando;Davizon-Castillo, Pavel;Page, Grier P;Kanias, Tamir
• 通讯作者: Kanias, Tamir

The prevalence and demographic determinants of blood donors receiving testosterone replacement therapy at a large USA blood service organization.

• DOI: 10.1111/trf.15754
• 发表时间: 2020-05
• 期刊: Transfusion
• 影响因子: 2.9
• 作者: Hazegh K;Bravo MD;Kamel H;Dumont L;Kanias T
• 通讯作者: Kanias T