Bioluminescence imaging of atherosclerosis associated reactive oxygen species

动脉粥样硬化相关活性氧的生物发光成像

• 批准号: 8058147
• 负责人: Reece Joseph Goiffon
• 金额: $ 2.81万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8058147
• 关键词: Anatomy; Animal Model; Apolipoprotein E; Arterial Fatty Streak; Arteriogram; Atherosclerosis; Biochemical; Biological; Biological Assay; Bioluminescence; Blood; Blood Cells; Blood specimen; Cardiovascular Diseases; Cause of Death; Cells; Cessation of life; Chemicals; Clinical; Complex; Cytolysis; Diagnosis; Disease; Early Diagnosis; Early treatment; Elements; Embolism; Endosomes; Enhancers; Environment; Enzyme-Linked Immunosorbent Assay; Enzymes; Goals; Hemorrhage; Hydrogen Peroxide; Hypochlorous Acid; Image; Imaging Techniques; Immunohistochemistry; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Laboratories; Life; Light; Luminol; Measurement; Measures; Medical; Methods; Microscopy; Modeling; Mus; Patients; Peer Review; Peroxidases; Phagocytes; Physiological; Plasma; Population; Predictive Value; Procedures; Process; Property; Radiation; Rattus; Reactive Oxygen Species; Reporting; Reproducibility of Results; Research; Research Project Grants; Risk; Role; Rupture; Sampling; Secondary Prevention; Serum; Signal Transduction; Simulate; Sodium; Solutions; Source; Species Specificity; Specificity; Stenosis; Stimulus; Superoxides; System; Techniques; Testing; Thrombosis; Time; Tissues; Trees; United States; Variant; Visible Radiation; Whole Blood; adverse outcome; analog L; base; combat; cost; digital; fluorescence imaging; fluorophore; imaging modality; imaging probe; improved; in vivo; inhibitor/antagonist; luciferin; macrophage; molecular imaging; mortality; neutrophil; next generation; oxidation; quantum; research study; response; scaffold; squaraine; tool

DESCRIPTION (provided by applicant): Cardiovascular disease (CVD) has consistently been the leading cause of death in the United States for nearly a century. Adverse outcomes of CVD usually result from rupturing atherosclerotic plaques. The most effective strategy for combating CVD is secondary prevention: diagnosis and treatment of subclinical disease. Non-invasively assessing atherosclerotic inflammation, the cause of plaque instability, is an important element of this strategy. Traditional imaging modalities cannot assess plaque vulnerability and thus are of limited predictive value. In contrast, molecular imaging visualizes the anatomy together with the biochemical signature of plaques. Current molecular imaging techniques used for CVD have setbacks, e.g., high cost and inherent risk to the patient. Bioluminescence imaging (BLI) does share these problems: oxidation-sensitive probes such as luminol and its derivative L-012 emit harmless, visible-spectrum light upon contact with reactive oxygen species (ROS) produced by phagocytes. These phagocytes are enriched in the arterial walls of CVD patients, mainly in and around inflamed plaques, and directly contribute to their rupture. The goal of this proposal is to develop BLI to detect ROS produced by these cells in animal models, thus allowing atherosclerotic inflammation to be imaged in vivo. This goal has been focused into three Specific Aims: In Aim 1, the chemiluminescence properties of three known BLI probes will be analyzed for use in single- cell bioluminescence microscopy (BLM). First, each probe will be tested for ROS specificity and chemical signal enhancement potential. The reproducibility of these findings in cellular environments will be tested with phagocytes isolated from animal models. Optimized conditions determined by these experiments will be used to visualize individual phagocytes with BLM as they initiate the ROS-synthesis cascade. In Aim 2, BLI will be used to measure inflammation of animal models both ex vivo and in vivo. First, BLI will be evaluated as an inexpensive alternative to current clinical assays for serum myeloperoxidase, a soluble enzyme marker for CVD and that creates the most potent physiological ROS, hypochlorous acid. Next, luminol derivative L-012 will be used to image atherosclerosis in apolipoprotein E-deficient mice. BLI signal will be correlated with plaque inflammation as quantified by immunohistochemistry. Aim 3 consists of a plan to synthesize new BLI probes that have improved properties for use in vivo, namely red-shifted emission spectra that are capable of penetrating deeper tissues. The proposed conjugates, oxazine and squaraine derivates, will be evaluated against current compounds in experiments similar to those outlined in Aims 1 and 2. The scaffolds of these conjugates will allow for a large number of variants to be synthesized with the possibility of becoming next-generation BLI probes. PUBLIC HEALTH RELEVANCE: Cardiovascular disease is the top cause of mortality in the United States, responsible for more than 1 out of every 3 deaths. This research project will develop techniques to make arterial plaques glow with visible light which doctors can photograph with highly sensitive digital cameras. This could allow earlier diagnosis and treatment and thus save more lives, all without any harmful radiation or highly invasive procedures.

描述（由申请人提供）：近一个世纪以来，心血管疾病（CVD）一直是美国的首要死因。 CVD 的不良后果通常是由于动脉粥样硬化斑块破裂造成的。对抗CVD最有效的策略是二级预防：亚临床疾病的诊断和治疗。非侵入性评估动脉粥样硬化炎症（斑块不稳定的原因）是该策略的重要组成部分。传统的成像方式无法评估斑块的脆弱性，因此预测价值有限。相比之下，分子成像将斑块的解剖结构和生化特征可视化。目前用于 CVD 的分子成像技术存在缺陷，例如成本高且对患者存在固有风险。生物发光成像 (BLI) 确实存在这些问题：鲁米诺及其衍生物 L-012 等氧化敏感探针在与吞噬细胞产生的活性氧 (ROS) 接触时会发出无害的可见光谱光。这些吞噬细胞在CVD患者的动脉壁中富集，主要是在发炎斑块内和周围，并直接导致其破裂。该提案的目标是开发 BLI 来检测动物模型中这些细胞产生的 ROS，从而对动脉粥样硬化炎症进行体内成像。这一目标集中在三个具体目标上： 在目标 1 中，将分析三种已知 BLI 探针的化学发光特性，以用于单细胞生物发光显微镜 (BLM)。首先，将测试每个探针的 ROS 特异性和化学信号增强潜力。这些发现在细胞环境中的重现性将用从动物模型中分离的吞噬细胞进行测试。这些实验确定的优化条件将用于在单个吞噬细胞启动 ROS 合成级联时用 BLM 可视化它们。 在目标 2 中，BLI 将用于测量离体和体内动物模型的炎症。首先，BLI 将被评估为当前血清髓过氧化物酶临床检测的廉价替代品，髓过氧化物酶是 CVD 的可溶性酶标记物，可产生最有效的生理 ROS（次氯酸）。接下来，鲁米诺衍生物 L-012 将用于对载脂蛋白 E 缺陷小鼠的动脉粥样硬化进行成像。 BLI 信号将与通过免疫组织化学定量的斑块炎症相关。 目标 3 包括合成新 BLI 探针的计划，这些探针具有改进的体内使用特性，即能够穿透更深组织的红移发射光谱。所提出的缀合物、恶嗪和方酸菁衍生物将在类似于目标 1 和 2 中概述的实验中针对当前化合物进行评估。这些缀合物的支架将允许合成大量变体，并有可能成为下一代 BLI 探针。 公共卫生相关性：心血管疾病是美国死亡的首要原因，每 3 人死亡中就有 1 人以上死于心血管疾病。该研究项目将开发使动脉斑块发出可见光的技术，医生可以用高灵敏度数码相机拍摄这些可见光。这可以实现更早的诊断和治疗，从而挽救更多的生命，而且无需任何有害的辐射或高度侵入性的手术。