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

基本信息

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
项目类别：
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

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

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项目摘要

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.

项目摘要镰状细胞病（SCD）是美国最常见的遗传血液学疾病。这 SCD中器官功能障碍的病理生理范围超出了异常的流变学和溶血，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中血管封闭（VO）的重要生化触发因素。活性氧（ROS）增加可以通过血小板和白细胞激活以及微血管功能障碍导致VO。无论氧化应激的药理减少可防止VO并改善微血管灌注。该建议的目的是使用最先进的微血管灌注和分子成像技术再加上先进的血浆生化分析，以测试两个有希望的抗氧化剂与降低SCD中的ROS有关：（a）N-乙酰囊这（NAC）和（b）apocynin。在 AIM 1，将研究SCD和控制小鼠的城镇鼠模型。非侵入性微血管肾脏和骨骼肌肉的对比增强超声（CEU）的灌注成像催眠后的条件将在预处理6周的小鼠中使用NAC，Apocynin或媒介物进行。参数灌注分析（微血管血容量和通量率）和色散建模将被使用更好地表征了导致SCD模型或治疗效果异常流动的血管事件。在AIM 2中，激活的Von Willebrand因子（VWF）和血小板的CEU分子成像以及PET分子氧化应激的成像将用于进一步评估负责的微血管分子事件 NAC或偶像性的异常灌注或治疗效果。在这两个目标中，高级LC-MS/MS 对于无偏的代谢组学并针对VWF的氧化修饰及其主要调节性鉴定将进行蛋白酶（ADAMTS-13）以及其他关键ROS，以评估药物作用。在AIM 3中，CEU 肢体骨骼肌，肾脏和心肌的灌注成像将在试验临床试验中进行测试以下假设，即减少NAC氧化应激可改善患者的微血管灌注与SCD。该试验被设计为与NAC或安慰剂的双重交叉，每次6周。结果该建议将解决我们对SCD病理生物学知识的关键差距，并提供有关新的治疗干预措施。它还将为PI提供丰富的研究培训计划（包括多学科指导和咨询团队，全面的课程和职业发展机构可用的资源）将奠定基础，成为一个独立的调查员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学疾病中的微血管病理生理。