Cluster Imaging of Emerging Biomarker Networks in Wound Healing

伤口愈合中新兴生物标志物网络的聚类成像

基本信息

批准号：
7693156
负责人：
Matthias Nahrendorf
金额：
$ 40.58万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7693156
关键词：
Address Algorithms Anatomy Apolipoprotein E Arterial Fatty Streak Arteries Biological Biological Markers Blood CX3CL1 gene Cell physiology Cells Classification Collagen Comorbidity Coronary Cytokine Receptors Data Data Set Detection Disease Equilibrium Exhibits Extracellular Matrix Flow Cytometry Fluorescence Fluorochrome Generations Genomics Goals Gold Healed Heart failure Histology Image Imaging Techniques Impaired wound healing Infarction Inflammation Inflammatory Integrins Investigational Therapies Lead Left Ventricular Remodeling Ligation Link Magnetic Resonance Imaging Maps Measurement Measures Microscopy Modeling Molecular Monitor Monocytosis Mus Myocardial Myocardial Infarction Optics Patients Pattern Peptide Hydrolases Peptides Performance Phase Photons Prevention Prodrugs Property Proteomics Reading Recruitment Activity Reporting Resolution Rupture Sampling Small Interfering RNA Staging Suspension substance Suspensions System Techniques Technology Therapeutic Therapeutic Intervention Time Tissues Transglutaminases Transillumination Validation Ventricular Western World Wound Healing angiogenesis base cardiovascular imaging chemokine receptor cost crosslink effective therapy forging healing in vivo inflammatory modulation macrophage molecular imaging monocyte mouse model nanoparticle new therapeutic target novel therapeutics prevent public health relevance reconstruction repaired research study therapeutic target tomography trafficking

项目摘要

DESCRIPTION (provided by applicant): Proteomic and genomic approaches provide increasingly comprehensive snapshots of biological networks. Often however, the molecular players in these networks are temporally and spatially very dynamic, and their precise mapping along the time-space continuum can become cost prohibitive or be sample limited. Noninvasive molecular imaging has the potential to longitudinally interrogate specific cellular and molecular hubs in vivo. Until recently, most macroscopic imaging techniques have been limited to display only one molecular player at any given time. We have developed quantitative 4-channel FMT-CT (Q4-FMT) to simultaneously map small "clusters" of biological targets in vivo. The technology is based on reconstruction of optical photons in transillumination geometry and fusion of 3D fluorescence data-sets with anatomic CT. We have recently adapted reconstruction algorithms to different wavelengths, effectively yielding 4 separate read-out channels. Here we propose to biologically validate and apply Q4-FMT for non-invasive interrogation of a biomarker network in wound healing models following myocardial infarction. We will use a set of imaging agents that interrogate key wound healing biomarkers: a) fluorescent nanoparticles to probe phagocytic activity, b) activatable optical agents to measure protease activity, c) tagged transglutaminase peptide substrates to report on extracellular matrix crosslinking, and d) integrin-targeted nanoparticles reporting on angiogenesis. First, systematic phantom experiments will address system performance in all channels, channel crosstalk, sensitivity, and precision of fluorochrome quantitation and image fusion. Second, in vivo imaging results will be quantitated and justaxposed to results from traditional, accepted gold standards such as flow cytometric analysis of cell suspensions from infarcts, genomic and proteomic data, and to immunoreactive histology. Finally, we will use Q4- FMT to assess therapeutic implications of monocyte subset recruitment and function on the above biomarkers during myocardial infarction in a mouse model of coronary ligation. This is important because timely detection of impaired healing would allow to intervene therapeutically to prevent heart failure. Using lipidoid-delivered siRNA we will silence CCR2. This experimental therapy targets the chemokine receptor that governs recruitment of inflammatory Ly-6Chi but not that of reparative Ly6Clo monocytes, while we will image infarct healing serially with Q4-FMT. We hypothesize that effects of modulating inflammatory monocyte recruitment can be monitored by Q4-FMT in realtime, enhances myocardial repair, and provides novel therapeutic strategies for infarct patients. PUBLIC HEALTH RELEVANCE: Our goal is to establish and validate Q4-FMT, which combines simultaneous 4-channel fluorescence molecular tomography with anatomical CT imaging, allowing to image a "cluster" of biomarkers simultaneously. We propose to validate Q4-FMT against established ex vivo gold standards and to apply it to study tissue repair after myocardial infarction to identify novel therapeutic targets for enhancement of myocardial repair and prevention of heart failure.

描述（由申请人提供）：蛋白质组学和基因组方法提供了越来越全面的生物网络快照。但是，这些网络中的分子播放器在时间和空间上是非常动态的，并且它们沿时间空间连续体的精确映射可能会变得越来越高，或者被限制为限制。非侵入性分子成像具有在体内纵向询问特定的细胞和分子轮毂的潜力。直到最近，大多数宏观成像技术都受到限制，在任何给定时间仅显示一个分子播放器。我们已经开发了定量的4通道FMT-CT（Q4-FMT），以同时绘制体内生物学靶标的小“簇”。该技术基于透射几何形状中光光子的重建以及与解剖CT的3D荧光数据集的融合。我们最近将重建算法调整为不同的波长，有效地产生了4个单独的读出通道。在这里，我们建议在心肌梗塞后生物学验证并应用Q4-FMT在伤口愈合模型中对生物标志物网络的无创询问。我们将使用一组询问关键伤口愈合生物标志物的成像剂：a）荧光纳米颗粒来探测吞噬活性，b）可激活的光学剂以测量蛋白酶的活性，c）标记的经凝集露核酶肽蛋白酶肽底物以报道蜂窝状互联蛋白的细胞外链接，并报告整合性的nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan nan。首先，系统的幻影实验将解决所有通道，通道串扰，敏感性以及荧光体定量和图像融合精度的系统性能。其次，体内成像结果将被定量，并被视为传统的，可接受的黄金标准的结果，例如对梗塞，基因组和蛋白质组学数据的细胞悬浮液的流式细胞仪分析以及免疫反应性组织学。最后，我们将使用Q4-FMT评估在冠状动脉结扎小鼠模型中，在心肌梗死期间，单核细胞子集募集和功能对上述生物标志物的治疗意义。这很重要，因为及时检测受损的愈合将允许治疗以防止心力衰竭进行干预。使用lipidoid传递的siRNA，我们将使CCR2沉默。这种实验疗法靶向趋化性LY-6CHI募集的趋化因子受体，而不是修复性LY6CLO单核细胞的趋化因子受体，而我们将用Q4-FMT串行梗死愈合。我们假设调节炎症单核细胞募集的影响可以通过Q4-FMT实时监测，增强心肌修复，并为梗死患者提供新颖的治疗策略。公共卫生相关性：我们的目标是建立和验证Q4-FMT，该Q4-FMT将同时的4通道荧光分子断层扫描与解剖CT成像结合在一起，从而可以同时对生物标志物的“群集”成像。我们建议验证Q4-FMT针对既定的离体金标准标准，并将其应用于心肌梗死后研究组织修复，以鉴定新的治疗靶标，以增强心肌修复和预防心力衰竭。