Fluorescent and Chemiluminescent Cyclooctyne Probes for Imaging Glycosylation

用于糖基化成像的荧光和化学发光环辛炔探针

• 批准号: 8461418
• 负责人: Yili Shi
• 金额: $ 1.72万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2013-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8461418
• 关键词: Affect; Animal Model; Azides; Behavior; Binding; Biological Models; Cell Line; Cell surface; Chemicals; Chinese Hamster; Chinese Hamster Ovary Cell; Copper; Ensure; Exhibits; Flow Cytometry; Fluorescence; Fluorescence Resonance Energy Transfer; Future; Generations; Goals; Image; Imagery; Imaging Device; Label; Malignant Neoplasms; Measures; Neoplasm Metastasis; Noise; Ovary; Parents; Polysaccharides; Property; Reaction; Research; Residual state; Resolution; Sensitivity and Specificity; Sialic Acids; Sorting - Cell Movement; Surface; System; Technology; Therapeutic; Transferase; Translating; analytical tool; cancer cell; cellular imaging; chemical reaction; design; effectiveness measure; glycosylation; imaging probe; improved; in vivo; inhibitor/antagonist; innovation; mutant; prognostic; sugar; tumor; tumor progression

DESCRIPTION (provided by applicant): Cancer cells display unusually glycosylated cell surfaces that are often correlated to cancer behavior. However, the question of why and when cancer cells exhibit aberrant glycosylation remains largely unanswered because current analytical tools cannot observe rapid glycosylation changes. I plan to develop an imaging platform able to reveal cancer glycosylation with both spatial and temporal resolution. The proposed strategy will use the chemical reaction of a cyclooctyne probe (copper-free click reaction) with an unnatural cell surface azide-labeled sugar to attach a visualizing agent to the cancer cell. The innovation of the new probes is sensitivity and specificity for azide imaging; the probes were designed to only form the visualizing agent after reaction with azide. This will increase imaging sensitivity because off-target binding and residual probe will not contribute to background noise. Two generations of cyclooctyne probes will be developed with two distinct visualizing agents: a fluorescent smart probe (Aim 1) and a chemiluminescent probe (Aim 2). The fluorescent cyclooctyne probe will be used to quantify sialic acid content on the surface of Chinese hamster ovary cells (Aim 3). The chemiluminescent cyclooctyne probe will be used to measure the effectiveness of a known sialyl transferase inhibitor with potential as an anti-cancer metastasis agent. The long-term goal of this project is to develop an imaging platform capable of elucidating how glycosylation affects cancer behavior in model organisms.

描述(申请人提供)：癌细胞显示异常糖基化的细胞表面，通常与癌症行为相关。然而，癌细胞为什么以及何时表现出异常糖基化的问题仍然在很大程度上没有答案，因为目前的分析工具不能观察到快速的糖基化变化。我计划开发一个能够同时以空间和时间分辨率显示癌症糖基化的成像平台。拟议的策略将使用环辛炔探针(无铜点击反应)与非天然细胞表面叠氮标记的糖的化学反应，将显像剂附着到癌细胞上。新探针的创新之处在于对叠氮化合物成像的敏感性和特异性； 探针被设计成仅在与叠氮反应后形成显影剂。这将提高成像灵敏度，因为偏离目标的结合和残留的探头不会对背景噪声造成影响。两代环辛炔探针将用两种不同的显像剂开发：一种荧光智能探针(AIM 1)和一种化学发光探针(AIM 2)。荧光环辛炔探针将用于定量中国仓鼠卵巢细胞表面唾液酸的含量(目标3)。化学发光的环辛炔探针将被用于测量一种已知的唾液酸基转移酶抑制剂的有效性，该抑制剂具有作为抗癌转移剂的潜力。该项目的长期目标是开发一个成像平台，能够阐明糖基化如何影响模式生物中的癌症行为。