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.

项目总结/摘要 控制释放装置有可能通过以下方式改善治疗结果和患者依从性 从而提供维持治疗窗内浓度的药物的延长的局部递送。 这种方法已被证明对许多应用有效，包括激素替代疗法，1 避孕药，2和癌症治疗，3但这些设备的发展，为其他应用 由于无法轻松研究它们在体内的行为，体外研究用于 评估从可生物降解的聚合物基质或水凝胶的治疗释放是简单和廉价的，但 由于水合作用的差异经常产生不代表体内释放动力学的结果， 缓冲、对流和酶活性。不幸的是， 可用于研究体内释放，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）是 昂贵、低通量，并且可能需要造影剂，其释放不代表 该项目旨在验证和优化一种高通量技术， 通过跟踪来自受控的药物的荧光标记蛋白的消耗来研究体内释放动力学， 释放装置。这种方法比现有的方法更快，成本更低，而且可能更安全 替代方案，还允许研究蛋白质特异性动力学，这可能与模型差异很大 proteins.简而言之，该技术将使用市售的基于简单的荧光标记试剂盒。 马来酰亚胺或N-羟基琥珀酰亚胺酯化学通过硫醇或伯胺标记蛋白质， 然后将荧光标记的蛋白质包封到聚合物装置（例如， 微粒、支架）并在体内植入或注射。一项纵向研究，包括定期体内 将进行荧光测量以评估控释制剂中剩余的蛋白质的量 随着时间的推移。为了验证这种方法的准确性，PET是一种低通量但高定量的方法 技术，将平行进行使用蛋白质，是双标记的荧光团和 放射性同位素荧光减少与衰变校正正电子减少之间的相关性 发射将被用来证明蛋白质含量和荧光之间的定量关系 信号了将使用各种荧光团、材料， 装置几何形状和植入部位，以减轻组织吸收，光漂白， 和荧光团pH敏感性，否则会对该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.

制造可填充的微粒和其他复杂的3D微观结构。

• 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