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）：我们将通过纳米级成像探针的设计，创建和测试来开发近红外（NIR）图像引导的干预措施的工具。这项提出的努力将通过三个目标来实现。 AIM 1：用于成像肿瘤细胞的一组独特的纳米级NIR探针的设计，制造和表征。 AIM 2：实施纳米级探针目标指导的组件。 AIM 3：使用NIR显微镜生成肿瘤细胞的空间和时间图像图。 AIM 1将包括在波长范围为850至2200 nm的水溶性NIR量子点（QD）的创建。这将通过修改使用油酸在有机溶剂中稳定的市售胶体铅硫化物（PBS）QD来实现。油酸将被赋予QD疏水性的特定硫醇化分子代替。接下来，将使用两亲块共聚物封装QD，并且将通过部分交联外部块来锁定整体结构。通过电子显微镜以及吸光度和荧光光谱法实现表征。通过AIM 2，将通过连接到单链片段变量抗体（A33SCFV），进一步修改水溶性QD，该抗体（A33SCFV）识别A33表面蛋白，该抗体在95％的原发性和转移性结直肠癌中表达。一旦解决了QD上限化学，将优化共价共轭方法。 A33SCFV-QD共轭物的特异性和敏感性将通过AIM 3使用人类结肠癌细胞系SW1222通过对照实验，使用A33- nonexpress的人类结肠癌细胞系HT29进行对照实验。这些细胞将在玻璃底培养皿中生长，随后与各种浓度的SCFV-QD结合物一起孵育。经过不同的孵育时间，将使用配备有CCD摄像头（明亮场成像）和NIR检测器的Olymus-BX51荧光显微镜在下面洗涤和可视化细胞。将收集明亮场和NIR图像的覆盖图片，NIR图像分析将允许对各个CT抗原的高分辨率成像，从而可以随着时间的推移进行精确定位和定位。当前的图像引导工具集很大程度上取决于可见光光谱，该光谱受到其渗透深度的限制。 NIR探针将扩展渗透深度，为指导外科医生提供更好的标记。我们的努力符合美国国家生物医学成像研究所和生物工程研究所在加速生物医学技术通过整合物理，工程和生命科学的应用方面的目标。拟议的研究还与国家癌症研究所的使命保持一致，以推进治疗性分娩，并通过改善早期检测和诊断方法来抢占癌症。这项最初的工作将为进一步开发工具的技术基础提供技术基础，这些工具将帮助临床医生不仅诊断，而且更重要的是采用图像引导的疗法。