Cancer molecular analysis using quantum dots

使用量子点进行癌症分子分析

基本信息

批准号：
9076630
负责人：
Xiaohu Gao
金额：
$ 23.52万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-01 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9076630
关键词：
Address Affect Antibodies Antigens Biological Markers Biomedical Research Buffers Cells Chemicals Clinical Communication Complex Consumption Cultured Cells Custom DNA DNA Probes Development Diagnosis Diagnostic Diagnostic Procedure Disease Emerging Technologies Ensure Evaluation Event Functional disorder Gene Chips Generations Goals Health Heterogeneity Human Human body Image Imaging technology Immune Immunofluorescence Immunologic In Situ Individual Kinetics Label Libraries Malignant Neoplasms Masks Mass Spectrum Analysis Mediating Medicine Methodology Methods Microscopy Modification Molecular Molecular Analysis Molecular Profiling Molecular Target Mutation Optics Pathologic Processes Performance Phenotype Portraits Preparation Process Production Property Protein Microchips Proteomics Protocols documentation Quantum Dots Reagent Reporter Reporting Research Resolution Reverse Transcriptase Polymerase Chain Reaction Route Sampling Semiconductors Signal Pathway Signal Transduction Specificity Specimen Staging Staining method Stains Systems Biology Technical Expertise Techniques Technology Time Tissues Translations Tumor Biology Tumor Markers Two-Dimensional Gel Electrophoresis Work accurate diagnosis antibody engineering anticancer research antigen antibody binding base body system cancer biomarkers cancer cell cell fixing clinical Diagnosis clinical practice cost effective flexibility hybrid antibody imaging biomarker innovation instrumentation neoplastic cell next generation novel optical imaging outcome forecast prognostic protein expression response screening self assembly success targeted treatment technology development therapy development tool tumor heterogeneity weapons

项目摘要

DESCRIPTION (provided by applicant): Comprehensive molecular characterization of individual cells is instrumental for understanding cell pathophysiology and our ability to diagnose and control the progression of complex pathological processes, such as cancer. In principle, tagging each biomolecule with a unique reporter probe and detecting its localization with high sensitivity at sub-cellular resolution can achieve this goal. Yet, none of the conventional biomedical techniques can stand up to the challenge, suffering from a limitation in the number of molecular targets that can be analyzed simultaneously, providing limited single cell information, and often utilizing qualitative rather than quantitative analysis. Translation of novel nanoparticl-based imaging technologies into biomedical research and clinical diagnostics promises to provide powerful tools for addressing phenotypic heterogeneity of tumors, opening access to studying low-abundance events often masked or completely erased by batch processing, and elucidating cancer biomarker signatures for accurate diagnosis and targeted therapy. To deliver on this promise, we recently invented a multicolor multicycle molecular profiling technology based on cyclic imaging of tens to hundreds of tumor biomarkers in single cells with optical imaging resolution (Nat. Commun. 2013 & Nat. Protocols 2013). Built on this work and another advance we made on DNA strand mediated displacement for fast and multicycle analysis (JACS 2011), we propose to explore and develop a new generation of multicolor multicycle proteomic technology with fast yet mild destaining condition. Our approach takes advantage of simple bioconjugation and self-assembly mechanisms for straightforward probe preparation, hybrid antibody-DNA probes for encoding of molecular targets, the unique optical properties of quantum dots for quantitative and multicolor imaging, and the mildness and fast kinetics of DNA toehold- mediated displacement. Unique combination of these features (which are supported by our recent preliminary results) yields simple, but powerful technology that should enable molecular characterization of individual cancer cells in situ, while requiring no specialized technical skills or significant modifications to common staining methodology and imaging instrumentation. As a result, if successful, the proposed QD technology should be readily applicable for a wide range of applications, in particular examination of molecular signaling pathways in cancer cells, evaluation of response to therapy, and development of multiplexed diagnostic and prognostic panels, thus promising to produce a substantial impact in cancer research and clinical practice.

描述（由申请人提供）：单个细胞的全面分子表征有助于理解细胞病理生理学和我们的诊断能力原则上，用独特的报告探针标记每个生物分子并以亚细胞分辨率高灵敏度检测其定位可以实现这一目标。能够应对挑战，同时分析的分子靶点数量有限，提供的单细胞信息有限，并且经常利用基于纳米颗粒的新型成像技术转化为生物医学研究和临床诊断。承诺提供强大的解决肿瘤表型异质性的工具，开放研究经常被批量处理掩盖或完全擦除的低丰度事件，并阐明癌症生物标志物特征以实现准确诊断和靶向治疗为了实现这一承诺，我们最近发明了一种多色多周期分子。基于光学成像分辨率的单细胞中数十至数百个肿瘤生物标志物的循环成像的分析技术（Nat. Commun. 2013 和 Nat. Protocols 2013 建立在这项工作和另一项工作的基础上）。鉴于我们在用于快速和多循环分析的 DNA 链介导的置换方面取得的进展（JACS 2011），我们建议探索和开发具有快速而温和脱色条件的新一代多色多循环蛋白质组技术，我们的方法利用简单的生物共轭和自组装。直接探针制备的机制、用于编码分子靶标的混合抗体-DNA探针、用于定量和多色成像的量子点的独特光学特性，以及DNA立足点的温和和快速动力学-这些特征的独特组合（我们最近的初步结果支持）产生了简单但强大的技术，可以对单个癌细胞进行原位分子表征，同时不需要专门的技术技能或对常见染色方法进行重大修改和因此，如果成功，所提出的 QD 技术应该可以轻松应用于广泛的应用，特别是癌细胞中分子信号传导途径的检查、治疗反应的评估以及多重诊断和预后组合的开发。，从而有望产生重大影响在癌症研究和临床实践中。