Fluorescence-based molecular imaging of in vivo release kinetics

基于荧光的体内释放动力学分子成像

• 批准号: 9429542
• 负责人: Kevin James McHugh
• 金额: $ 6.12万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9429542
• 关键词: Amines; Animal Model; Antibody titer measurement; Antigens; Attention; Behavior; Biocompatible Materials; Biological Assay; Blood; Blood Vessels; Bovine Serum Albumin; Buffers; Chemistry; Clinic; Collection; Contraceptive Agents; Contrast Media; Convection; Device or Instrument Development; Devices; Diabetes Mellitus; Drops; Drug Delivery Systems; Emission-Computed Tomography; Emulsions; Encapsulated; Esters; Evaluation; Fluorescence; Fluorescent Probes; Formulation; Geometry; Glycolates; Hormone replacement therapy; Hydration status; Hydrogels; Image; Imaging Techniques; Implant; In Vitro; Kinetics; Label; Libraries; Longitudinal Studies; Magnetic Resonance Imaging; Maleimides; Malignant Neoplasms; Measurement; Measures; Medical Device; Methods; Modeling; Mus; Nature; Noise; Patients; Performance; Periodicity; Photobleaching; Photons; Polymers; Polymethyl Methacrylate; Positron; Positron-Emission Tomography; Property; Proteins; Radiation; Radioisotopes; Research Personnel; Research Technics; Resolution; Safety; Signal Transduction; Site; Solvents; Sulfhydryl Compounds; Surface; Techniques; Technology; Tetanus Toxoid; Therapeutic; Time; Tissue Engineering; Tissues; Toxic effect; Translations; Vaccination; Vascular Endothelial Growth Factors; Vascularization; Work; X-Ray Computed Tomography; Zirconium; arm; base; biodegradable polymer; biomaterial compatibility; cancer therapy; caprolactone; clinically relevant; compliance behavior; controlled release; cost; density; design; enzyme activity; evaporation; fluorescence imaging; fluorophore; high throughput screening; imaging modality; implantable device; implantation; improved; in vivo; in vivo optical imaging; interest; molecular imaging; non-invasive optical imaging; pre-clinical; pre-clinical research; preclinical evaluation; preclinical study; premature; quantum; scaffold; sebacic acid; single photon emission computed tomography; subcutaneous; therapeutic protein; therapy outcome; vaccine delivery

PROJECT SUMMARY/ABSTRACT Controlled release devices have the potential to improve therapeutic outcomes and patient compliance by providing prolonged, localized delivery of drugs that maintain concentrations within the therapeutic window. This approach has proven effective for a number of applications including hormone replacement therapy,1 contraceptives,2 and cancer therapy,3 yet the development of these devices for additional applications continues to be hindered by an inability to easily study their behavior in the body. In vitro studies used to assess therapeutic release from biodegradable polymer matrices or hydrogels are simple and inexpensive, but frequently yield results that are not representative of in vivo release kinetics due to differences in hydration, buffering, convection, and enzyme activity. Unfortunately, non-invasive preclinical imaging modalities that could be used to study in vivo release such as magnetic resonance imaging (MRI), computed tomography (CT), positron emission tomography (PET), and single-photon emission computed tomography (SPECT) are expensive, low-throughput, and may require contrast agents whose release is not representative of the therapeutic protein of interest.4 This project aims to validate and optimize a high-throughput technique for studying in vivo release kinetics by tracking the depletion of fluorescently labeled proteins from controlled release devices. This approach is faster, considerably less expensive, and potentially safer than existing alternatives and also allows for the study of protein-specific kinetics which could differ greatly from model proteins. In brief, this technique will use commercially available fluorescent labeling kits based on simple maleimide or N-hydroxysuccinimide ester chemistry to label proteins via thiols or primary amines, respectively.5 Fluorescently labeled proteins will then be encapsulated into polymeric devices (e.g. microparticles, scaffolds) and implanted or injected in vivo. A longitudinal study consisting of periodic in vivo fluorescence measurements will be performed to assess the amount of protein remaining in controlled release devices over time. To validate the accuracy of this approach, PET, a low-throughput but highly quantitative technique, will be performed in parallel using proteins that are double labeled with fluorophores and a radioisotope. Correlation between the decrease in fluorescence and decrease in decay-corrected positron emission will be used to demonstrate the quantitative relationship between protein content and fluorescent signal. The robustness of fluorescence-based release will be evaluated using various fluorophores, materials, device geometries, and implant sites in order to mitigate the influence of tissue absorbance, photobleaching, and fluorophore pH-sensitivity that could otherwise negatively impact the accuracy of this approach.

项目摘要/摘要 受控释放设备有可能通过 提供长时间的局部递送药物，这些药物在治疗窗口内保持浓度。 事实证明，这种方法对包括激素替代疗法在内的许多应用有效，1 避孕药2和癌症疗法，3但这些设备开发用于其他应用 由于无法轻松研究其在体内的行为而继续阻碍。体外研究曾经 评估可生物降解的聚合物基质或水凝胶的治疗释放是简单且便宜的，但 由于水合的差异，经常产生的结果无法代表体内释放动力学的结果， 缓冲，对流和酶活性。不幸的是，非侵入性临床前成像方式 可用于研究体内释放，例如磁共振成像（MRI），计算机断层扫描 （CT），正电子发射断层扫描（PET）和单光子发射计算机断层扫描（SPECT）为 昂贵，低通量，并且可能需要对比剂，而对比剂的释放不代表 4该项目旨在验证和优化高通量技术 通过跟踪受控的荧光标记的蛋白质的耗竭来研究体内释放动力学 释放设备。这种方法比现有的速度更快，便宜且可能更安全 替代方法，还允许研究蛋白质特异性动力学，这可能与模型有很大不同 蛋白质。简而言之，该技术将使用基于简单的市售荧光标签套件 马来酰亚胺或N-羟基糖酸酯化学化学，通过硫醇或原代胺标记蛋白质， 5然后将荧光标记的蛋白质封装在聚合设备中（例如， 微粒，支架）并植入或注入体内。一项由周期性体内的纵向研究 将进行荧光测量，以评估受控释放中剩余的蛋白质量 随着时间的推移设备。为了验证这种方法的准确性，PET是一种低通量但高度定量的 技术，将使用用荧光团双重标记的蛋白质并行进行 放射性同位素。荧光减少与衰减校正正电子的减少之间的相关性 排放将用于证明蛋白质含量与荧光之间的定量关系 信号。将使用各种荧光团，材料， 设备几何形状和植入地点，以减轻组织吸光度，光漂白的影响， 以及荧光团pH值，否则可能会对这种方法的准确性产生负面影响。

项目成果

Fabrication of fillable microparticles and other complex 3D microstructures.

• DOI: 10.1126/science.aaf7447
• 发表时间: 2017-09-15
• 期刊: Science (New York, N.Y.)
• 作者: McHugh KJ;Nguyen TD;Linehan AR;Yang D;Behrens AM;Rose S;Tochka ZL;Tzeng SY;Norman JJ;Anselmo AC;Xu X;Tomasic S;Taylor MA;Lu J;Guarecuco R;Langer R;Jaklenec A
• 通讯作者: Jaklenec A