Detection of plaque based macrophages with light

用光检测基于斑块的巨噬细胞

• 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
• 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

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）： 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）已被证明可以准确成像薄纤维帽和大脂质核心。我们现在提出了一种方法，将检测巨噬细胞在脆弱的斑块与OCT患者在导管插入术的时间。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，我们可以首次使用相位敏感OCT与刺激激光耦合来检测巨噬细胞的存在。在这种方法中，由基于斑块的巨噬细胞吞噬的光学纳米颗粒通过由于应用脉冲激光而引起的热膨胀和弛豫而进入纳米（nm）运动，并且这种运动用相敏OCT系统检测。我们还开发了用于这种方法的新型光学纳米颗粒，其由最大波长为700-800 nm的光激发，以防止激光与竞争性斑块成分（如血红蛋白、脂质和动脉壁）相互作用，所述竞争性斑块成分最大吸收500-600 nm的光。由于它们的多面表面，我们创造了术语“纳米糖”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，我们提出了以下具体目标：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、金和葡聚糖量的纳米糖，以实现强NIR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巨噬细胞对纳米糖的特异性摄取，而不是培养中的内皮细胞和平滑肌细胞。1.3我们将静脉注射奈米糖至动脉硬化的兔子，并利用萤光显微镜进行详细的组织学检查，以确定奈米糖在斑块中摄取的最小可侦测剂量与特异性。具体目标#2-使用纳米糖对兔动脉粥样硬化斑块中巨噬细胞进行离体和体内OCT成像。2.1我们将纳米糖静脉注射到动脉粥样硬化兔中，对离体主动脉组织进行OCT成像，并通过RAM-11识别巨噬细胞和纳米糖的高光谱成像将这些图像与组织学进行比较。2.2我们将纳米糖静脉注射到动脉粥样硬化兔中，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-纳米糖的排泄和毒性。3.1我们将描述纳米糖的排泄途径（肝脏与肾脏、分布器官和血液半衰期）。3.2我们将通过血液化学（肝和肾损伤、嗜酸性粒细胞生成的证据）和死后检查证明家兔中不存在器官毒性。这些研究也将与纳米技术表征实验室合作进行（见支持函）。 公共卫生关系： 开发一种基于对比剂-光的联合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，因此应用所提出的可以对有破裂风险的斑块进行成像的技术可以减少心脏病发作、中风和外周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提供基础，以在大量患者人群中验证拟议方法。