Spectroscopic Quantification of Ligand Binding in Vivo

体内配体结合的光谱定量

• 批准号: 8310098
• 负责人: JOHN B WEAVER
• 金额: $ 21.55万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8310098
• 关键词: Advanced Malignant Neoplasm; Affinity; Animal Model; Animals; Antibodies; Back; Binding; Biological Assay; Blood Circulation; Blood Vessels; Cancer Center; Cell Culture System; Cell Culture Techniques; Cell surface; Cells; Cellularity; Clinical Treatment; Collection; Computer software; Custom; Data; Development; Diagnostic; ERBB2 gene; Effectiveness; Engineering; Evaluation; Fluorescence; Fluorescence Spectroscopy; Goals; Image; In Vitro; Injection of therapeutic agent; Instruction; Intercellular Fluid; Investigation; Iron; Ligand Binding; Ligands; Magnetic Resonance Imaging; Magnetism; Malignant Neoplasms; Measurement; Measures; Methodology; Methods; Microscopic; Modeling; Motion; Movement; Mus; Nanotechnology; Optical Methods; Optics; Patients; Performance; Phagocytes; Phase; Positioning Attribute; Process; Recording of previous events; Relative (related person); Resources; Signal Transduction; Site; Specificity; Spectrum Analysis; Study of magnetics; System; Techniques; Technology; Time; Tissues; Translating; Treatment outcome; Ultrasonography; Validation; Variant; Vesicle; Work; cancer imaging; cancer therapy; effective therapy; feeding; fluorophore; imaging modality; improved; in vivo; innovation; insight; instrumentation; interstitial; nanoparticle; new technology; novel; novel strategies; optical imaging; particle; pressure; programs; ratiometric; success; targeted delivery; therapeutic effectiveness; therapy design; tissue phantom; tomography; tool; tumor; uptake

PROJECT SUMMARY (See instructions); The aim of this project is to develop new methodology to quantitatively image targeted magnetic nanoparticle (mNP) distribution in vivo in each microscopic compartment to provide mechanistic insights into where targeted mNPs collect and how targeted delivery might be improved. The project will combine two novel synergistic methods, optical ratiometric fluorescence spectroscopy (OFS) and magnetic spectroscopy of mNP Brownian motion (MSB), to quantify bulk concentration uptake and then quantify the level of specific binding to the target ligands in vivo, focusing initially on HER2/neu targeted mNPs which can be internalized when bound. Both measurement techniques proposed here have been uniquely developed at Dartmouth and we have considerable existing resources and expertise that can be leveraged. The global distribution and the bound fraction will be measured with OFS by injection of non-targeted and targeted mNP decorated with different fluorophores. Through quantification of the ratio of uptake, the fraction of bound agent can be measured. There is a large resource of in vivo fluorescence tomography apparatus at Dartmouth, allowing measurement with MicroCT, ultrasound, high field MRI and whole body MRI scanners, and substantial expertise in software, hardware and animal studies have developed in the past decade to allow routine application of this approach. The project will adapt existing hardware/software to couple with magnetic spectroscopy and study the mNP developed by this program. MSB has the ability to measure the binding energies ofthe mNP directly in vivo by evaluating the mNP Brownian motion. Currently no methods are able to do this. The average binding energy for the mNP at each position will be measured. The average binding energy and the bound faction from OFS will provide estimates ofthe fraction of mNP bound to the cell surface and in vesicles. In vivo and ex vivo studies will be used to validate the specificity of the binding, as well as the localization relative to the MSB signal. This technology platform is a fundamentally new, unique way to quantify mNP binding in vivo, and the tools developed here can be utilized in a wide range of mNP targeting assessments. When developed within the program, we can make the technology available for other mNP targeting constructs to help assess the relative success of targeting in vivo, and track the binding status over time. The ultimate value of this technology would be to allow longitudinal tracking of binding in vivo in clinical treatments, thereby providing a custom binding dosimeter for use in patient-specific treatments.

项目摘要（请参阅说明）； 该项目的目的是开发新的方法来定量图像靶向磁性纳米颗粒 （MNP）每个显微镜中的体内分布 有针对性的MNP收集以及如何改善目标交付。该项目将结合两本小说 协同方法，光学比率荧光光谱（OFS）和磁光谱法 MNP Brownian运动（MSB），以量化大量浓度吸收，然后量化特定水平 在体内与靶配体结合，最初集中在HER2/NEU靶向MNP上 绑定时。此处提出的两种测量技术都是在达特茅斯和 我们拥有可利用的大量现有资源和专业知识。 全球分布和约束分数将通过注射非目标和 有针对性的MNP用不同的荧光团装饰。通过定量摄取比率，分数 可以测量结合代理的。在体内荧光断层扫描设备上有大量资源 达特茅斯（Dartmouth 在过去的十年中，在软件，硬件和动物研究方面发展了丰富的专业知识 允许常规应用此方法。该项目将使现有的硬件/软件与 磁光谱和研究该程序开发的MNP。 MSB具有通过评估MNP直接在体内测量MNP的结合能 布朗运动。目前，没有任何方法能够执行此操作。每个MNP的平均结合能 位置将进行测量。 OFS的平均结合能量和约束派系将提供 MNP分数结合到细胞表面和囊泡中的估计值。体内和体内研究将是 用于验证结合的特异性以及相对于MSB信号的定位。 该技术平台是一种从根本上量化体内MNP绑定的新的独特方法， 这里开发的工具可以在多种MNP靶向评估中使用。当内部开发时 该计划，我们可以使技术可用于其他MNP靶向构造，以帮助评估 靶向体内靶向的相对成功，并随时间跟踪结合状态。最终价值 技术将允许在临床治疗中对体内结合的纵向跟踪，从而提供 定制结合剂量表用于特定于患者的治疗。