Multi-spectral quantum dot-based retinal imaging of molecular expression in vivo

基于多光谱量子点的体内分子表达视网膜成像

基本信息

批准号：
7192344
负责人：
Frederick R Haselton
金额：
$ 19.17万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-03-01 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7192344
关键词：
Acridine Orange Adhesions Animal Disease Models Animal Model Antibodies Arteries Asthma Atherosclerosis Binding Biological Biological Markers Blood Blood Circulation Blood Platelets Blood Vessels CD31 Antigens Cell Adhesion Molecules Cell Culture System Cell membrane Cell surface Cells Characteristics Code Complex Cultured Cells Cytotoxic T-Lymphocytes Detection Development Diabetes Mellitus Diagnostic Diagnostic Procedure Disease Disease Progression Dyes E-Selectin Early identification Endothelial Cells Endothelium Etiology Event Extravasation Eye Fluorescence Fluorescence Microscopy Fluorescent Probes Image Imaging Techniques Immune response In Vitro Individual Inflammation Inflammation Mediators Inflammatory Inflammatory Response Injection of therapeutic agent Intercellular adhesion molecule 1 Invasive Label Laboratories Leukocyte Rolling Leukocyte Trafficking Leukocytes Malignant Neoplasms Mediator of activation protein Methodology Methods Microcirculation Modality Modeling Molecular Molecular Probes Monitor Monoclonal Antibodies Nanotechnology Noise Numbers Optics Pathology Pharmaceutical Preparations Play Polymers Population Process Property Protein Engineering Quantum Dots Radiolabeled Rattus Relative (related person)Research Resistance Resolution Retina Retinal Role Signal Transduction Site Source Specificity Speed Streptozocin Surface Surface Antigens T-Lymphocyte Techniques Technology Therapeutic Therapeutic Intervention Time Tissues Tracer Treatment Efficacy Tumor Antigens Up-Regulation Validation Vascular Cell Adhesion Molecule-1 Veins Work angiogenesis antibody conjugate base cell type charge coupled device camera concept design desire diabetic diabetic rat fluorescence imaging fluorescence microscope fluorophore immunogenicity in vivo intercellular cell adhesion molecule interest light intensity molecular dynamics molecular imaging monocyte nanocrystal nanoprobe neutrophil optical imaging quantum radiotracer research study residence response size surface coating time use tissue preparation tool uptake vascular inflammation

项目摘要

DESCRIPTION (provided by applicant): Undesirable provocation of the inflammatory response is a detrimental feature of numerous diseases such as diabetes, atherosclerosis, and asthma and thus impacts a significant population. Current strategies utilized to identify key markers of inflammatory disease at the cellular and molecular level have often been centered on in vitro cell culture studies or analyses of tissue excised from animal models of disease. In this proposal we describe the development of a platform for the real-time, in vivo analysis of multiple cellular and molecular mediators of inflammation in diseased animal models using quantum dot nanocrystals and retinal fluorescence microscopy. Our design differs from previous approaches to study inflammation in vivo in that it is enabled by the harnessing of unique and highly-desirable optical properties conferred to quantum dots. These properties include higher quantum efficiency relative to conventional dyes, a resistance to fading, narrow and size-tunable emission spectra all excitable by one wavelength, and amenability to surface engineering of proteins and polymers for optimum stability and targeting. The second enabling technology is our laboratory expertise in in vivo retinal fluorescence imaging which provides unique continuous optical accessibility to the in vivo vasculature. To validate the potential of our proposed technique, in Aim 1 we seek to optimize an imaging modality / nanoprobe design to simultaneously detect the therapeutically-significant biomarkers of inflammation ICAM-1, VCAM-1, PECAM-1, and E-selectin, and to observe their relative molecular expression levels in the retinal vasculature in a rat model of diabetes. Furthermore, we seek to adapt the probe/fluorescence imaging design established in Aim 1 to track multiple leukocyte subsets in real time in vivo in the same animal model. Our preliminary work demonstrates the potential of this technique for the specific labeling and in vivo detection of the endothelial surface markers ICAM-1, VCAM-1 and PECAM-1 in vivo, as well as moving, in vivo-labeled neutrophils in the retinal circulation, with high spatial and temporal resolution and high signal to background ratios. A highlight of our preliminary studies was the first in vivo validation of VCAM-1 and ICAM-1 upregulation in diabetes. This technology has the potential to elucidate complex, cellular and molecular events as they occur in real-time not only in inflammation, but in other diseases such as cancer and ocular disorders as well.

描述（由申请人提供）：炎症反应的不良挑衅是许多疾病（例如糖尿病，动脉粥样硬化和哮喘）的有害特征，从而影响了大量人群。当前用于鉴定细胞和分子水平上炎症性疾病关键标记的策略通常以体外细胞培养研究或从动物疾病动物模型中切除的组织分析为中心。在此提案中，我们描述了使用量子点纳米晶体和视网膜荧光显微镜在患病动物模型中对实时，体内分析炎症多个细胞和分子介质的平台的发展。我们的设计与以前研究体内炎症的方法有所不同，因为它可以利用与量子点赋予的独特且高度可取的光学特性来实现。这些特性包括相对于常规染料的较高的量子效率，一种对褪色，狭窄且可调的发射光谱的抗性，所有波长都可以通过一个波长进行激发，以及蛋白质和聚合物表面工程的适应性，以实现最佳稳定性和靶向。第二种促成技术是我们在体内视网膜荧光成像方面的实验室专业知识，它为体内脉管系统提供了独特的连续光学可访问性。为了验证我们提出的技术的潜力，在AIM 1中，我们试图优化成像方式 /纳米探针设计，以同时检测炎症ICAM-1，VCAM-1，PECAM-1，PECAM-1和E-选择性的治疗性生物标志物，并观察其相对分子表达在视网膜脉络膜中的相对分子表达水平。此外，我们试图适应AIM 1中建立的探针/荧光成像设计，以在同一动物模型中实时跟踪多个白细胞子集。我们的初步工作证明了该技术对内皮表面标志物的特定标记和体内检测的潜力，体内ICAM-1，VCAM-1和PECAM-1和PECAM-1在体内以及在视网膜循环中移动，体内标记的中性粒细胞，具有高空间和时间分辨率和高信号和高度信号和高度信号。我们初步研究的亮点是糖尿病中VCAM-1和ICAM-1上调的体内验证。该技术不仅在炎症中，而且在其他疾病（例如癌症和眼部疾病）中实时发生，因此有可能阐明复合物，细胞和分子事件。