Impaired release of antiadhesive ATP from stored RBCs: a novel transfusion lesion

储存的红细胞中抗粘附 ATP 的释放受损：一种新的输血损伤

基本信息

批准号：
8285703
负责人：
TIMOTHY J MCMAHON
金额：
$ 23.55万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8285703
关键词：
Adhesions Adhesives Anemia Benefits and Risks Biochemical Biological Assay Biological Preservation Blood Blood Vessels Blood flow Cardiac Surgery procedures Cell Adhesion Cell surface Cessation of life Clinical Clinical Trials Critical Illness Data Disease Effectiveness Elements Endothelial Cells Endothelium Equilibrium Erythrocyte Transfusion Erythrocytes Exposure to Frequencies Functional disorder Goals Health Human Iloprost Impairment In Vitro Infusion procedures Integrins Lesion Link Lung Malaria Mediating Mediator of activation protein Medicine Modeling Morbidity - disease rate Mus Outcome Oxygen Patients Phosphoric Acid Esters Prostaglandins I Regulation Rejuvenation Respiratory Failure Risk Role S-Nitrosothiols Signal Transduction Signaling Molecule Testing Transfusion Translations Vasodilator Agents Vasomotor Work adhesion receptor analog design diabetic improved in vitro Assay in vivo innovation intercellular cell adhesion molecule mouse model novel patient population prevent respiratory sickling uptake

项目摘要

DESCRIPTION (provided by applicant): Impaired release of anti-adhesive ATP from stored RBCs: a novel transfusion lesion Transfusion of red blood cells (RBCs) is a frequently administered and costly therapy. Yet in some patient populations - the critically ill, for example clinical outcomes after transfusion are disappointing, and may worsen as a function of the duration of RBC storage. Biochemical and functional changes in banked RBCs are well established, but few or none are credibly linked to the adverse clinical outcomes in transfusion recipients. We have demonstrated that storage progressively impairs the ability of human RBC to release ATP, and that this impairment promotes the adhesion of RBCs to endothelial cells (ECs) in vitro, and sequestration of RBCs in the lung in vivo, with worsened oxygenation in a novel model of transfusion in mice. This novel proadhesive RBC "storage lesion" is mediated by the RBC-surface adhesion receptor ICAM-4 (LW) and endothelial ¿v¿3 integrin, and is prevented by co-infusion of an authentic ATP analog. We hypothesize that the storage- induced deficiency in ATP release from transfused human RBCs contributes to pathophysiology in the host through the locally injurious sequelae of endothelial adhesion of RBCs, compromising O2 uptake and delivery. We will test this hypothesis by accomplishing these Specific Aims: 1) Determine the mechanism by which released ATP inhibits RBC adhesion to endothelial cells. We will extend our novel findings that pharmacological inhibition of panx1 (that blocks RBC ATP release) also promotes RBC adhesion by studying the ability of panx1-deficient RBCs to export ATP and their propensity to adhere to endothelium. We will also determine the mechanism of anti-adhesive effects of RBC-derived ATP by testing the influence of specific purinergic antagonists and the role of ATP-sensitive activation of the relevant adhesion receptors. 2) Determine the effectiveness of strategies to a) replete ATP in stored RBCs and b) promote or mimic ATP-release in vitro and upon transfusion in vivo. We will achieve this aim in endothelial cell adhesion assays in vitro and in the murine human-RBC transfusion model we have recently developed. The influence of post-storage ATP repletion ("rejuvenation") of RBCs will be determined in vitro and in vivo, and the influence of co-infusion of ATP analogs will also be determined. These novel studies are expected to accelerate our nascent understanding of how the adhesion of normal RBCs is modulated by the release of ATP, a novel function likely to have broader significance, as in the increased endothelial adhesion of sickle, diabetic and malaria-infected RBCs. The innovative approach of examining the adhesion of both human (and mouse) RBCs in a murine model is expected to accelerate the translation of our findings toward clinical investigation in transfusion medicine. The results of these studies are expected to inform the rational design of strategies to improve the risk-benefit balance for RBC transfusion in critically ill and other anemic patients. PUBLIC HEALTH RELEVANCE: Transfusion of red blood cells (RBCs) for anemia fails to provide a benefit in many patient groups, and may raise the risk in some of death and other problems such as respiratory failure. This project focuses on storage- induced declines in the RBC's ability to release the signaling molecule ATP, and the functional consequences of these progressive changes. In particular, we investigate the link between impaired ATP export by stored RBCs and the harmful tendency for RBCs to stick to blood vessels rather than flowing through smoothly in order to pick up oxygen in the lungs and deliver it to the body. We then investigate whether these deficiencies can be corrected in human RBCs, and in transfused mice.

描述（由适用提供）：从储存的RBC中释放抗粘合剂ATP的受损：红细胞（RBC）的新型输血病变输血（RBC）经常进行且昂贵的治疗。然而，在某些患者人群中 - 危重患病，例如输血后的临床结果令人失望，并且随着RBC储存持续时间的函数可能会更糟。银行RBC的生化和功能变化已建立得很好，但是很少或没有与输血接受者的不良临床结果相关。我们已经证明，存储逐渐损害了人类RBC释放ATP的能力，并且这种损害促进了RBC在体外对内皮细胞（ECS）的粘附，以及在体内肺中RBC的隔离，在小鼠中具有新颖的传输模型中，肺部有氧化剂。这种新型的原始RBC“储存病变”是由RBC表面粘附受体ICAM-4（LW）和内皮emential»v¿3整合素介导的，并通过对真实的ATP Analog共感染来阻止。我们假设，储存引起的ATP释放中的ATP缺乏通过局部损害RBC的内皮粘附的后遗症促进了宿主的病理生理，从而损害了O2的摄取和递送。我们将通过完成这些特定目的来检验这一假设：1）确定释放ATP抑制RBC对内皮细胞的粘附的机制。我们将扩展我们的新发现，即PANX1的药理学抑制（阻止RBC ATP释放）还通过研究Panx1缺乏率RBC出口ATP的能力及其遵守内皮的承诺，从而促进了RBC的粘附。我们还将通过测试特定的嘌呤能拮抗剂的影响以及相关粘附受体对ATP敏感激活的作用来确定RBC衍生ATP的抗粘附作用的机制。 2）确定策略对a）替换储存的RBC中的ATP的有效性，b）促进或模仿体外输血后的ATP释放。我们将在体外和最近开发的鼠类RBC输血模型中实现这一目标。 RBC的存储后ATP复制（“复兴”）的影响将在体外和体内确定，并且还将确定ATP类似物共连接的影响。预计这些新的研究将加速我们对正常RBC粘附的敏捷理解，即通过释放ATP的释放来调节，ATP的释放是一种新型功能，可能具有更广泛的显着性，因为在镰状，糖尿病和疟疾感染的RBC的内皮粘合度增加中。在鼠模型中检查人（和小鼠）RBC的粘附的创新方法预计将加速我们对输血医学临床投资的发现的转化。这些研究的结果预计将为策略的合理设计提供了改善RBC的风险效益平衡的策略。病和其他贫血患者。公共卫生相关性：红细胞（RBC）对贫血的输血无法为许多患者群体提供好处，并且可能会增加某些死亡和其他问题（例如呼吸衰竭）的风险。该项目的重点是RBC释放信号分子ATP的能力以及这些渐进性变化的功能后果的关注。特别是，我们研究了储存的RBC的ATP导出受损与RBC粘在血管上的有害趋势之间的联系，而不是平稳地流过血管，以便在肺部捡起氧气并将其输送到体内。然后，我们研究是否可以在人类RBC和输血小鼠中纠正这些缺陷。