Role of RBC NO and ATP in Sickle Vasculopathy

红细胞 NO 和 ATP 在镰状血管病中的作用

• 批准号: 8392230
• 负责人: TIMOTHY J MCMAHON
• 金额: $ 37.37万
• 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: 2012-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8392230
• 关键词: Abnormal Red Blood Cell; Acute; Adhesions; Adhesives; Adverse effects; Affect; Anti-Inflammatory Agents; Anti-inflammatory; Binding; Biological Models; Biology; Blood Cells; Blood Transfusion; Blood Vessels; Blood flow; Cell Adhesion; Cell Communication; Cell-Cell Adhesion; Cells; Cellular biology; Chronic; Complex; Data; Development; Disease; Endothelium; Erythrocytes; Event; Failure; Functional disorder; Gases; Goals; Hypoxia; In Vitro; Individual; Inflammation; Inflammatory; Knowledge; Lead; Leukocytes; Life; Lung; Maintenance; Measurable; Mediating; Membrane; Modality; Morbidity - disease rate; Mus; Nitric Oxide; Normal tissue morphology; Organ; Pain; Pathology; Perfusion; Phenotype; Physiology; Procedures; Publishing; Pulmonology; Receptor Signaling; Regulation; Reporting; Research; Research Personnel; Respiratory physiology; Role; S-Nitrosothiols; Sickle Cell; Sickle Cell Anemia; Sickle Hemoglobin; Signal Pathway; Signal Transduction; Testing; Therapeutic; Tissues; Transfusion; Vascular Diseases; Vasodilation; Work; adhesion receptor; base; hemodynamics; improved; in vivo; innovation; leukocyte activation; meetings; mortality; mouse model; novel therapeutic intervention; prevent; response; restoration; sensor; sickling; vasoconstriction; Abnormal Red Blood Cell; Acute; Adhesions; Adhesives; Adverse effects; Affect; Anti-Inflammatory Agents; Anti-inflammatory; Binding; Biological Models; Biology; Blood Cells; Blood Transfusion; Blood Vessels; Blood flow; Cell Adhesion; Cell Communication; Cell-Cell Adhesion; Cells; Cellular biology; Chronic; Complex; Data; Development; Disease; Endothelium; Erythrocytes; Event; Failure; Functional disorder; Gases; Goals; Hypoxia; In Vitro; Individual; Inflammation; Inflammatory; Knowledge; Lead; Leukocytes; Life; Lung; Maintenance; Measurable; Mediating; Membrane; Modality; Morbidity - disease rate; Mus; Nitric Oxide; Normal tissue morphology; Organ; Pain; Pathology; Perfusion; Phenotype; Physiology; Procedures; Publishing; Pulmonology; Receptor Signaling; Regulation; Reporting; Research; Research Personnel; Respiratory physiology; Role; S-Nitrosothiols; Sickle Cell; Sickle Cell Anemia; Sickle Hemoglobin; Signal Pathway; Signal Transduction; Testing; Therapeutic; Tissues; Transfusion; Vascular Diseases; Vasodilation; Work; adhesion receptor; base; hemodynamics; improved; in vivo; innovation; leukocyte activation; meetings; mortality; mouse model; novel therapeutic intervention; prevent; response; restoration; sensor; sickling; vasoconstriction

DESCRIPTION (provided by applicant): Vasoconstriction, blood cell adhesion, and inflammation are each potentially devastating events in sickle cell disease (SCD). Sickle red cells (SS RBCs) demonstrate complex membrane and biologic abnormalities. Nitric oxide (NO) delivered by RBCs is both critical in the maintenance of vasodilation and a potent anti-inflammatory agent. ATP is also released by RBCs and signals increases in blood flow to meet O2 demand, typically by enhancing NO synthesis. While normal (AA) RBCs act as a hypoxia sensor by releasing both ATP and bioactive nitric oxide (NO), leading to NO-dependent vasodilation, SS RBCs are deficient in both content and ability to release both NO and ATP. The ability of SS RBCs to adhere to the endothelium and to activate leukocytes as well as other cells, along with their failure to induce pulmonary vasodilation, may result in part from their deficiencies in membrane-bound bioactive S-nitrosothiol (SNO) and ATP. Our preliminary data show that loading SS RBCs with NO/SNO down-regulates SS RBC adhesion, S RBC-stimulated leukocyte adhesion, and vaso-occlusion in vivo and modulates the vasoconstrictive pulmonary phenotype. Inhibition of ATP release by RBCs also induces RBC adhesion and pulmonary vasoconstriction. In SCD, abnormal vascular tone, cell adhesion, leukocyte activation, and inflammation are all believed to contribute to the pathophysiology of vaso-occlusion, which is central to both painful crises and acute and chronic organ damage. Thus, our central hypothesis is that NO and ATP deficits in SS RBCs directly contribute to both SS RBC adhesion, S RBC-induced activation of leukocytes, and pulmonary vasoconstriction. We further postulate that restoration of NO and ATP content in either SS or transfused (stored) AA RBCs will also improve some of the measurable adverse effects of SS RBCs in the lung. To test our hypothesis and progress toward achieving improved therapies for SCD, we have combined the efforts of investigators with expertise in SS RBC biology, transfusion medicine, and pulmonary physiology in order to 1) Determine the influence of NO/SNO- and ATP-repletion on pulmonary hemodynamics and gas exchange in isolated lungs and intact mice transfused with SS RBCs alone or in combination with strategies that modulate RBC adhesive events; 2) Test whether NO and ATP repletion of SS RBCs or stored AA RBCs can relieve vaso-occlusion in a mouse model in vivo; and 3) Determine the effect of SS RBC NO and ATP on the activation of RBC adhesion receptors in vitro, the signaling pathways involved, and RBC activation of leukocytes. Our long-term goal is to improve vascular tone, cell adhesion, and cell activation in SCD by identifying remediable SS RBC abnormalities and thereby reduce vaso-occlusion. This work will allow development of new therapeutic approaches to prevent and control vaso-occlusion and tissue damage in SCD.

描述（由申请人提供）：血管收缩，血细胞粘附和炎症是镰状细胞疾病（SCD）中的毁灭性事件。镰状红细胞（SS RBC）表现出复杂的膜和生物学异常。 RBC递送的一氧化氮（NO）对于维持血管舒张和有效的抗炎剂至关重要。 RBC还释放了ATP，并且信号增加了血流以满足O2需求，通常通过增强无合成。虽然正常（AA）RBC通过释放ATP和生物活性一氧化氮（NO）来充当缺氧传感器，导致NO依赖性血管舒张，但SS RBC在含量和释放NO和ATP的内容方面都缺乏。 SS RBC粘附在内皮上并激活白细胞以及其他细胞的能力，以及它们未能诱导肺血管舒张的能力，可能会导致其在膜结合的生物活性S-硝基硫醇（SNO）和ATP中的缺陷。我们的初步数据表明，无/sno的SS RBC在体内加载SS RBC，ss rbc粘附，S rbc刺激的白细胞粘附力和血管批准在体内并调节血管收缩的肺部表型。 RBC对ATP释放的抑制还会诱导RBC粘附和肺部血管收缩。在SCD中，据信血管张力异常，细胞粘附，白细胞激活和炎症都促进了血管牙合的病理生理学，这对于疼痛危机和急性危机和急性和慢性器官损害至关重要。因此，我们的中心假设是，SS RBC中的NO和ATP缺陷直接导致SS RBC粘附，S RBC诱导的白细胞激活和肺部血管收缩。我们进一步假设，SS或输血（存储）AA RBC中NO和ATP含量的恢复还将改善肺中SS RBC的某些可测量的不良影响。为了检验我们的假设和进步，我们将研究人员的努力与SS RBC生物学，输血医学和肺部生理学方面的专业知识相结合，以便为了1）确定NO/SNO-SNO-SNO-RE-和ATP重复的影响与单独的肺部和气体交换的影响，以使SS-Remantion对孤立的血液动力和气体交换，以使SS型肺部和气体在孤立的血液动力和气体中的组合，将其与SS型液体交换组合。调节RBC粘合剂事件； 2）测试SS RBC或存储的AA RBC的NO和ATP补充是否可以在体内减轻小鼠模型中的血管酸度； 3）确定SS RBC NO和ATP对体外RBC粘附受体激活的影响，涉及的信号通路以及白细胞的RBC激活。我们的长期目标是通过鉴定可补充的SS RBC异常，从而减少血管含量，从而改善SCD中的血管张力，细胞粘附和细胞活化。这项工作将允许开发新的治疗方法，以预防和控制SCD中的血管咬合和组织损伤。