Oxidative stress in vaso-occlusion pathophysiology of sickle cell disease

镰状细胞病血管闭塞病理生理学中的氧化应激

• 批准号: 9333429
• 负责人: Melinda D Wu
• 金额: $ 17.44万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-16 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9333429
• 关键词: ADAMTS; Acute; Address; Adhesions; Advisory Committees; Animals; Antioxidants; Biochemical; Blood Platelets; Blood Vessels; Blood Volume; Blood flow; Career Mobility; Cell Adhesion Molecules; Cells; Chronic; Clinical Research; Clinical Trials; Complex; Contrast Media; Crossover Design; Data; Disease; Disease model; Disintegrins; Endothelial Cells; Endothelium; Enzymes; Equilibrium; Erythrocytes; Event; Exercise; Foundations; Functional disorder; Gene-Modified; Genetic; Glutathione Reductase; Goals; Hematological Disease; Hemoglobin; Hemolysis; Human; Hypoxia; Image; Imaging Techniques; Impairment; In Situ; Inflammation; Inflammatory; Injury; Institution; Kidney; Knowledge; Lead; Limb structure; Lipids; Measurement; Measures; Mediating; Mentorship; Metalloproteases; Microbubbles; Microvascular Dysfunction; Modeling; Modification; Molecular; Mus; Muscle; Myocardium; NADP; NOS3 gene; Nitric Oxide; Organ; Oxidases; Oxidative Stress; Oxygen; P-Selectin; Pain; Patients; Peptide Hydrolases; Perfusion; Peroxonitrite; Pharmaceutical Preparations; Pharmacology; Phytochemical; Placebos; Plasma; Platelet Activation; Play; Positron-Emission Tomography; Process; Production; Proteins; Pulmonary Vascular Resistance; Reactive Oxygen Species; Recombinants; Recruitment Activity; Reperfusion Injury; Research Personnel; Research Training; Resources; Rest; Rheology; Role; Serum Markers; Sickle Cell Anemia; Site; Skeletal Muscle; Stress; Techniques; Testing; Therapeutic; Thrombosis; Thrombospondins; Tissues; Training Programs; Ultrasonography; United States; Vascular resistance; Vasoconstrictor Agents; Vasodilator Agents; Xanthine Oxidase; acetovanillone; blood flow measurement; cleavage factor; contrast enhanced; design; disorder control; glutathione peroxidase; improved; in vivo; inhibitor/antagonist; insight; leukocyte activation; liquid chromatography mass spectrometry; metabolomics; microPET; molecular imaging; mouse model; multidisciplinary; novel; novel therapeutic intervention; oxidation; perfusion imaging; polymerization; pre-clinical; prevent; skills; targeted treatment; therapeutic target; treatment effect; treatment strategy; vascular abnormality; vasomotion; von Willebrand Factor

PROJECT SUMMARY Sickle cell disease (SCD) is the most common genetic hematologic disorder in the United States. The pathophysiology of organ dysfunction in SCD extends beyond abnormal rheology and hemolysis, and includes abnormal vascular function, thrombosis, and inflammation. There is increasing evidence that oxidative stress is an important biochemical trigger for vaso-occlusion (VO) in SCD. Increased reactive oxygen species (ROS) can lead to VO through platelet and leukocyte activation, and microvascular dysfunction. Whether pharmacologic reduction in oxidative stress prevents VO and improves microvascular perfusion is not known. The goal of this proposal is to use state-of-the-art microvascular perfusion and molecular imaging techniques together with advanced plasma biochemical analysis to test mechanism and therapeutic impact of two promising anti-oxidants implicated in reducing ROS in SCD: (a) N-acetylcystine (NAC), and (b) apocynin. In Aim 1, the Townes murine model of SCD and control mice will be studied. Non-invasive microvascular perfusion imaging with contrast-enhanced ultrasound (CEU) of the kidney and skeletal muscle under normoxic and post-hypoxic conditions will be performed in mice pre-treated for 6 weeks with NAC, apocynin, or vehicle. Parametric perfusion analysis (microvascular blood volume and flux rate) and dispersion modeling will be used to better characterize the vascular events responsible for abnormal flow in the SCD model or treatment effect. In Aim 2, CEU molecular imaging of activated von Willebrand Factor (vWF) and platelets, and PET molecular imaging of oxidative stress will be used to further assess the microvascular molecular events responsible for abnormal perfusion or treatment effect from the NAC or apocynin. In both of these Aims, advanced LC-MS/MS for unbiased metabolomics and targeted identification of oxidative modification of VWF and its main regulatory protease (ADAMTS-13), as well as other key ROS, will be performed to evaluate drug effect. In Aim 3, CEU perfusion imaging of limb skeletal muscle, kidney, and myocardium will be performed in a pilot clinical trial testing the hypothesis that reducing oxidative stress with NAC improves microvascular perfusion in patients with SCD. This trial is designed as a double cross-over with NAC or placebo for 6 weeks each. The results of this proposal will address a critical gap in our knowledge of the pathobiology of SCD and provide insight into new therapeutic interventions. It will also provide the PI with a rich research training program (including multidisciplinary mentorship and advisory teams, comprehensive coursework, and career advancement resources available at the institution) that will lay the foundation to become an independent investigator who is able to use advanced imaging and other analytic techniques in pre-clinical and clinical studies to examine microvascular pathophysiology in hematologic disease.

项目总结 镰状细胞病(SCD)是美国最常见的遗传性血液疾病。这个 SCD中器官功能障碍的病理生理学超越了异常流变学和溶血，包括 血管功能异常、血栓形成和炎症。越来越多的证据表明，氧化应激 是SCD血管闭塞(VO)的重要生化触发因素。活性氧(ROS)增加 可通过激活血小板和白细胞导致VO，并导致微血管功能障碍。是否 药物减轻氧化应激可预防VO并改善微血管灌注量，目前尚不清楚。 这项提议的目标是使用最先进的微血管灌注和分子成像技术 结合先进的血浆生化分析检测两种药物的作用机制及治疗效果 有望降低SCD中ROS的抗氧化剂：(A)N-乙酰半胱氨酸(NAC)和(B)罗布麻素。在……里面 目的1、建立Townes小鼠SCD模型，并与对照组小鼠进行比较。无创微血管 常氧状态下肾脏和骨骼肌的超声造影灌注成像 在用NAC、罗布麻素或赋形剂预处理6周的小鼠中，进行缺氧后的条件。 将使用参数灌注分析(微血管血容量和流量)和弥散模型 以更好地表征导致SCD模型中异常血流或治疗效果的血管事件。 在目标2中，激活的von Willebrand因子(VWF)和血小板的CEU分子成像，以及PET分子 氧化应激成像将被用于进一步评估微血管分子事件 由NAC或Apoynin引起的异常血流灌注或治疗效果。在这两个目标上，先进的LC-MS/MS 无偏代谢组学和靶向鉴定VWF及其主要调控因子的氧化修饰 将使用蛋白水解酶(ADAMTS-13)以及其他关键ROS来评估药物效果。在AIM 3中，CEU 肢体骨骼肌、肾脏和心肌的灌注成像将在一项试点临床试验中进行。 NAC降低氧化应激改善患者微血管灌注率的假设检验 使用SCD。这项试验设计为双交叉，NAC或安慰剂各6周。结果是 这项建议将解决我们在SCD病理生物学知识方面的一个关键差距，并提供对 新的治疗干预措施。它还将为PI提供丰富的研究培训方案(包括 多学科指导和咨询团队、全面的课程工作和职业发展 该机构可用的资源)，这将为成为一名独立的调查员奠定基础 能够在临床前和临床研究中使用先进的成像和其他分析技术来检查 血液病的微血管病理生理学。