Targeted Near-Infrared Probes for Image-Guided Cancer Interventions

用于图像引导癌症干预的靶向近红外探针

• 批准号: 7409776
• 负责人: Diego A Rey
• 金额: $ 3.75万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7409776
• 关键词: Animals; Antibodies; Antibody Repertoire; Antigens; Area; Attention; Biological; Biological Preservation; Biological Sciences; Biomedical Engineering; Biomedical Technology; Cancer Intervention; Cancerous; Cell Line; Cells; Chemicals; Chemistry; Clinic; Collaborations; Colon Carcinoma; Colorectal Cancer; Complement; Contrast Media; Depth; Development; Diagnosis; Diagnostic; Early Diagnosis; Electron Microscopy; Encapsulated; Engineering; Equilibrium; Excision; Fluorescence Spectroscopy; Foundations; Future; GPA33 gene; Generations; Glass; Goals; Human; Image; Image Analysis; Imaging Device; Incubated; Institutes; Intervention; Invasive; Label; Laboratories; Malignant Epithelial Cell; Malignant Neoplasms; Maps; Membrane Proteins; Methods; Mission; Modification; National Cancer Institute; Oleic Acid; Oleic Acids; Operative Surgical Procedures; Organic solvent product; Other Therapy; Patients; Penetration; Physicians; Positioning Attribute; Quantum Dots; Range; Reagent; Research; Resolution; Sensitivity and Specificity; Site; Speed; Staging; Structure; Surgeon; Testing; Therapeutic; Time; Tissues; Tumor Antigens; Visible Radiation; Water; Work; base; bioimaging; cancer diagnosis; cancer imaging; cancer therapy; charge coupled device camera; colon cancer cell line; concept; copolymer; crosslink; design; detector; fluorescence microscope; image guided intervention; image guided therapy; imaging probe; infrared microscopy; lead sulfide; metastatic colorectal; nanoparticle; nanoscale; neoplastic cell; optical imaging; preempt; research study; success; tool; tool development

DESCRIPTION (provided by applicant): We will develop tools for near-infrared (NIR) image-guided interventions through the design, creation and testing of nanoscale imaging probes. This proposed effort will be accomplished through three aims. Aim 1: the design, fabrication and characterization of a unique set of nanoscale NIR probes for imaging tumor cells. Aim 2: the implementation of components for target directing of nanoscale probes. Aim 3: the generation of spatial and temporal image-based maps of tumor cells using NIR microscopy. Aim 1 will comprise the creation of water-soluble NIR quantum dots (QDs) that emit in the wavelength range from 850 to 2200 nm. This will be accomplished through modification of commercially available colloidal lead-sulfide (PbS) QDs which are stabilized in organic solvents using oleic acid. Oleic acid will be replaced with specific thiolated molecules rendering the QDs hydrophobic. Next, amphiphilic block copolymers will be used to encapsulate QDs and the overall structure will be locked by partially cross-linking the outer block. Characterization will be achieved through electron microscopy together with absorbance and fluorescence spectroscopy. Through Aim 2, the water soluble QDs will then be further modified through conjugation to a single chain fragment variable antibody (A33scFv) that recognizes the A33 surface protein which is expressed in 95% of primary and metastatic colorectal cancers. A covalent conjugation method will be optimized once the QD capping chemistry has been resolved. The specificity and sensitivity of the A33scFv-QD conjugates will be investigated through Aim 3 using the human colon carcinoma cell line SW1222 with control experiments using the A33-nonexpressing human colon cancer cell line HT29. The cells will be grown in glass-bottom Petri dishes and subsequently incubated with various concentrations of ScFv-QD conjugates. After various incubation times, cells will be washed and visualized under using an Olymus-BX51 fluorescence microscope equipped with both a CCD camera (bright-field imaging) and a NIR detector. Overlaid pictures of bright field and NIR images will be collected and NIR image analysis will permit high resolution imaging of the respective CT antigens allowing for their precise positioning and localization over time. The current sets of image-guided tools largely depend on the visible light spectrum which is limited by its depth of penetration. NIR probes will extend penetration depths providing better markers for guiding the surgeon. Our efforts are in line with the goals of the National Institute of Biomedical Imaging and Bioengineering in accelerating the application of biomedical technologies through integrating the physical, engineering, and life sciences. The proposed study is also aligned with the mission of the National Cancer Institute in advancing therapeutic delivery and it's goal to preempt cancer through improvements in early detection and diagnostics. This initial work will provide the technical foundation for the further development of tools that will help clinicians not only diagnose but more importantly employ image-guided therapies.

描述（由申请人提供）：我们将通过设计、创建和测试纳米级成像探针来开发近红外（NIR）图像引导干预的工具。这项拟议的努力将通过三个目标来实现。目标 1：设计、制造和表征一套独特的纳米级近红外探针，用于肿瘤细胞成像。目标 2：实现纳米级探针目标导向组件。目标 3：使用近红外显微镜生成基于空间和时间图像的肿瘤细胞图。目标 1 将包括创建水溶性近红外量子点 (QD)，其发射波长范围为 850 至 2200 nm。这将通过对市售胶体硫化铅 (PbS) 量子点进行改性来实现，该量子点使用油酸在有机溶剂中稳定。油酸将被特定的硫醇分子取代，使量子点具有疏水性。接下来，两亲性嵌段共聚物将用于封装量子点，并通过部分交联外部嵌段来锁定整体结构。表征将通过电子显微镜以及吸光度和荧光光谱来实现。通过目标 2，水溶性量子点将通过与单链片段可变抗体 (A33scFv) 缀合来进一步修饰，该抗体可识别 A33 表面蛋白，该蛋白在 95% 的原发性和转移性结直肠癌中表达。一旦量子点封端化学得到解决，共价结合方法将得到优化。 A33scFv-QD 缀合物的特异性和敏感性将通过目标 3 使用人结肠癌细胞系 SW1222 进行研究，并使用不表达 A33 的人结肠癌细胞系 HT29 进行对照实验。细胞将在玻璃底培养皿中生长，随后与不同浓度的 ScFv-QD 缀合物一起孵育。经过不同的孵育时间后，将洗涤细胞并使用配备 CCD 相机（明场成像）和 NIR 检测器的 Olymus-BX51 荧光显微镜进行观察。将收集明场和 NIR 图像的重叠图片，NIR 图像分析将允许对各个 CT 抗原进行高分辨率成像，从而随着时间的推移对其进行精确定位和定位。当前的图像引导工具很大程度上依赖于可见光谱，而可见光谱受到其穿透深度的限制。 NIR 探头将扩大穿透深度，为指导外科医生提供更好的标记。我们的努力符合国家生物医学成像和生物工程研究所的目标，通过整合物理、工程和生命科学来加速生物医学技术的应用。拟议的研究也符合国家癌症研究所推进治疗交付的使命，其目标是通过改进早期检测和诊断来预防癌症。这项初步工作将为进一步开发工具提供技术基础，这些工具不仅可以帮助临床医生进行诊断，更重要的是可以帮助他们采用图像引导疗法。