Detection of plaque based macrophages with light

用光检测基于斑块的巨噬细胞

基本信息

批准号：
8259055
负责人：
MARC David FELDMAN
金额：
--
依托单位：
SOUTH TEXAS VETERANS HEALTH CARE SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8259055
关键词：
Aorta Arterial Fatty Streak Atherosclerosis Biological Biology Biomedical Engineering Blood Blood Chemical Analysis Blood Circulation Blood Vessels Cardiac Cardiac Catheterization Procedures Cardiovascular system Catheterization Cause of Death Cells Cerebrum Cessation of life Chemical Engineering Clinical Research Coin Collaborations Collection Colloids Contrast Media Coupled Data Detection Development Dextrans Dose Excretory function Flow Cytometry Fluorescence Future Goals Gold Growth Half-Life Hemoglobin Hepatic Histology Image Imaging Techniques Infiltration Intravenous Kidney Label Laboratories Lasers Lead Letters Light Lipids Liquid substance Liver Magnetic Resonance Angiography Matrix Metalloproteinases Mechanics Methodology Motion Myocardial Infarction Nanotechnology Optical Coherence Tomography Optics Organ Oryctolagus cuniculus Pathologic Pathway interactions Patients Peripheral Catheterization Peripheral Vascular Diseases Phase Photophobia Physicians Physiologic pulse Production Property Public Health Relaxation Research Resolution Risk Rupture Scientist Sensitivity and Specificity Shoulder Signal Transduction Smooth Muscle Myocytes Societies Specificity Stroke Surface System Techniques Time Tissues Toxic effect Translating Translations Ultrasonography Veterans abstracting acute coronary syndrome base cell type cellular imaging density design dextran eosinophil in vivo iron oxide light scattering macrophage member multidisciplinary nanocage nanometer nanoparticle nanorod nanoshell novel optical imaging particle patient population prevent programs receptor research study surface coating time use uptake

项目摘要

DESCRIPTION (provided by applicant): Project Summary/Abstract The pathologic features that predict atherosclerotic plaque rupture are large lipid collections, thinning of the fibrous cap, and infiltration of macrophages. Optical Coherence Tomography (OCT) has already been demonstrated to accurately image thin fibrous caps and large lipid cores. We now propose an approach which will detect macrophages in vulnerable plaque with OCT in patients at the time of catheterization. We demonstrate that by labeling plaque-based macrophages with intravenous gold nanoparticles we can detect the presence of macrophages for the first time using phase-sensitive OCT coupled with a stimulating laser. In this approach, optical nanoparticles engulfed by plaque-based macrophages are put into nanometer (nm) motion via thermal expansion and relaxation due to application of a pulsed laser, and this motion detected with a phase-sensitive OCT system. We have also developed novel optical nanoparticles for this approach which are excited by light maximally at a wavelength of 700-800 nm, to prevent laser interaction with competing plaque components such as hemoglobin, lipid, and arterial wall which maximally absorb light at 500-600 nm. Due to their multi-faceted surface, we have coined the term "nanorose" to describe these nanoparticles. To demonstrate cellular imaging of macrophages in intact plaque, we propose the following specific aims: Specific Aim # 1 - Specificity and sensitivity of nanoparticle (nanorose) uptake. 1.1 We will synthesize and fully characterize nanorose with controlled size, and gold and dextran amounts, to achieve strong NIR absorbance and specific cell (macrophage) uptake. 1.2 We will demonstrate specific uptake of nanorose by macrophages as opposed to endothelial and smooth muscle cells in culture via the use of flow cytometry. 1.3 We will inject nanorose iv into atherosclerotic rabbits, and perform detailed histology with fluorescence microcopy to determine both the minimal detectable dose and the specificity of nanorose uptake in plaque. Specific Aim # 2 - Ex vivo and in vivo OCT imaging of macrophages in rabbit atherosclerotic aortas with nanorose. 2.1 We will inject nanorose iv into atherosclerotic rabbits, perform OCT imaging of ex vivo aortic tissue, and compare these images with histology via RAM-11 identification of macrophages, and hyperspectral imaging of nanorose. 2.2 We will inject nanorose iv into atherosclerotic rabbits, repeat each of the histological experiments in section 2.1, but with in vivo OCT imaging. Specific Aim # 3 - Excretion and toxicity of nanorose. 3.1 We will characterize the excretion pathway of nanorose (hepatic vs. renal, organs of distribution, and blood half-life). 3.2 We will demonstrate in rabbits the absence of organ toxicity via blood chemistry for evidence of hepatic and renal damage, eosinophil production, and post-mortem examination. These studies will also be performed in collaboration with the Nanotechnology Characterization Laboratory (see letter of support). PUBLIC HEALTH RELEVANCE: Project Narrative Development of a combined contrast agent - light based approach to identify vulnerable plaques will have a significant impact on the public health. Inasmuch as atherosclerosis remains the leading cause of death in Western society, application of the proposed technique that can image plaques at risk for rupture can reduce the number of deaths from heart attack, stroke and peripheral vascular disease. Successful completion of the proposed research will provide a basis for expanded clinical studies to validate the proposed methodology in a large patient population.

描述（由申请人提供）：项目摘要/摘要预测动脉粥样硬化斑块破裂的病理特征是大量脂质聚集、纤维帽变薄和巨噬细胞浸润。光学相干断层扫描 (OCT) 已被证明可以对薄纤维帽和大脂质核心进行精确成像。我们现在提出了一种在患者插管时使用 OCT 检测易损斑块中巨噬细胞的方法。我们证明，通过用静脉内金纳米颗粒标记基于斑块的巨噬细胞，我们可以使用相敏 OCT 结合刺激激光首次检测巨噬细胞的存在。在这种方法中，由于脉冲激光的应用，被基于斑块的巨噬细胞吞噬的光学纳米颗粒通过热膨胀和松弛而进入纳米（nm）运动，并且用相敏OCT系统检测到这种运动。我们还为此方法开发了新型光学纳米粒子，其最大程度被波长为 700-800 nm 的光激发，以防止激光与竞争性斑块成分（例如血红蛋白、脂质和动脉壁）相互作用，这些成分最大吸收 500-600 nm 的光。纳米。由于它们的多面表面，我们创造了术语“纳米玫瑰”来描述这些纳米颗粒。为了证明完整斑块中巨噬细胞的细胞成像，我们提出以下具体目标：具体目标#1 - 纳米颗粒（纳米糖）摄取的特异性和敏感性。 1.1 我们将合成并充分表征具有受控尺寸、金和葡聚糖含量的纳米玫瑰，以实现强近红外吸收和特定细胞（巨噬细胞）摄取。 1.2 我们将通过使用流式细胞术证明培养物中的巨噬细胞（而不是内皮细胞和平滑肌细胞）对纳米玫瑰的特异性摄取。 1.3 我们将纳米玫瑰静脉注射到动脉粥样硬化兔子中，并通过荧光显微镜进行详细的组织学分析，以确定斑块中纳米玫瑰摄取的最小可检测剂量和特异性。具体目标#2 - 使用纳米玫瑰对兔动脉粥样硬化主动脉中的巨噬细胞进行离体和体内 OCT 成像。 2.1 我们将纳米玫瑰静脉注射到动脉粥样硬化兔子中，对离体主动脉组织进行 OCT 成像，并通过巨噬细胞的 RAM-11 识别和纳米玫瑰的高光谱成像将这些图像与组织学进行比较。 2.2 我们将纳米玫瑰静脉注射到动脉粥样硬化兔子中，重复第 2.1 节中的每个组织学实验，但使用体内 OCT 成像。具体目标#3 - 纳米玫瑰的排泄和毒性。 3.1 我们将描述纳米玫瑰的排泄途径（肝与肾、分布器官和血液半衰期）。 3.2 我们将通过血液化学来证明兔子不存在器官毒性，作为肝肾损伤、嗜酸性粒细胞产生和尸检的证据。这些研究还将与纳米技术表征实验室合作进行（参见支持信）。公共卫生相关性：项目叙述开发一种组合造影剂-基于光的方法来识别易损斑块将对公众健康产生重大影响。由于动脉粥样硬化仍然是西方社会死亡的主要原因，因此应用所提出的技术对有破裂风险的斑块进行成像可以减少心脏病发作、中风和周围血管疾病的死亡人数。拟议研究的成功完成将为扩大临床研究奠定基础，以在大量患者群体中验证拟议的方法。