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

• 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
• 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

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（由申请人提供）：储存的红细胞中抗粘附ATP释放受损：一种新型输血病变红细胞（RBC）的输血是一种经常使用且昂贵的治疗方法。然而，在一些患者群体中，例如重症患者，输血后的临床结果令人失望，并且可能随着RBC储存时间的延长而恶化。库存RBC的生化和功能变化已得到充分证实，但很少或没有与输血受者的不良临床结局相关。我们已经证明，存储逐渐损害人红细胞释放ATP的能力，这种损害促进红细胞的粘附内皮细胞（EC）在体外，和隔离的红细胞在肺中的体内，恶化的氧合在一个新的模型中的输血小鼠。这种新的促粘附红细胞"储存损伤"是由红细胞表面粘附受体ICAM-4（LW）和内皮细胞整合素3介导的，并通过共同输注一种真正的ATP类似物来预防。我们假设，储存诱导的输注人RBC ATP释放缺陷通过RBC内皮粘附的局部损伤性后遗症，损害O2摄取和递送，导致宿主的病理生理学。我们将通过实现以下具体目标来检验这一假设：1）确定释放的ATP抑制RBC粘附于内皮细胞的机制。我们将通过研究panx1缺陷型红细胞输出ATP的能力及其粘附于内皮的倾向来扩展我们的新发现，即panx1的药理学抑制（阻断RBC ATP释放）也促进RBC粘附。我们还将通过测试特异性嘌呤能拮抗剂的影响和相关粘附受体的ATP敏感性激活的作用来确定RBC衍生的ATP的抗粘附作用的机制。2)确定以下策略的有效性：a）在储存的RBC中补充ATP和B）促进或模拟体外和体内输血后ATP释放。我们将在体外内皮细胞粘附试验和我们最近开发的小鼠人红细胞输注模型中实现这一目标。将在体外和体内确定RBC的储存后ATP补充（"复原"）的影响，并且还将确定ATP类似物的共输注的影响。这些新的研究有望加速我们对正常红细胞的粘附如何通过ATP的释放来调节的初步理解，ATP是一种可能具有更广泛意义的新功能，如镰状、糖尿病和疟疾感染的红细胞的内皮粘附增加。在小鼠模型中检查人（和小鼠）RBC粘附的创新方法有望加速我们的研究结果向输血医学临床研究的转化。这些研究的结果有望为改善危重患者红细胞输注的风险-获益平衡策略的合理设计提供信息。 病患者和其他贫血患者。 公共卫生相关性：输注红细胞（RBC）治疗贫血在许多患者群体中无法提供益处，并且可能会增加一些死亡和其他问题（如呼吸衰竭）的风险。这个项目的重点是储存引起的红细胞释放信号分子ATP的能力下降，以及这些进行性变化的功能后果。特别是，我们研究了储存的红细胞受损的ATP输出与红细胞粘附在血管上而不是顺利流过血管以获取肺部氧气并将其输送到身体的有害趋势之间的联系。然后，我们研究这些缺陷是否可以在人红细胞和输血小鼠中得到纠正。