Sensitive and multiplexed analysis of cancer biomarkers with QD barcodes

使用 QD 条形码对癌症生物标志物进行灵敏的多重分析

• 批准号: 7740849
• 负责人: Xiaohu Gao
• 金额: $ 31.86万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-14 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7740849
• 关键词: Achievement; Biological; Biological Markers; Biosensor; Caliber; Cells; Chemicals; Chemistry; Clinical; Color; Complex; DNA; DNA Microarray Chip; DNA analysis; Data; Detection; Development; Diagnosis; Diagnostic; Disease; Dyes; Early Detection Research Network; Enzyme-Linked Immunosorbent Assay; Event; Fluorescence; Gene Expression Profiling; Generations; Genetic; Genomics; Goals; Image; Immobilization; Label; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Microfluidic Microchips; Molecular; Molecular Analysis; Molecular Profiling; Mutation; Names; Nanosphere; National Cancer Institute; Nucleic Acids; Oligonucleotides; Optics; Photobleaching; Polymers; Principal Investigator; Process; Property; Proteins; Proteomics; Quantum Dots; RNA; Reporter; Research; Resistance; Science; Screening for cancer; Semiconductors; Serum; Signal Transduction; Specimen; Surface; System; Techniques; Technology; Temperature; Therapeutic; Time; Tissues; Validation; absorption; anticancer research; base; copolymer; drug discovery; flexibility; improved; innovation; innovative technologies; instrumentation; light emission; melting; miniaturize; molecular assembly/self assembly; nanoparticle; nanoscale; new technology; novel; optical imaging; prognostic; programs; protein profiling; research study; seal; self assembly; single molecule; therapeutic target

DESCRIPTION (provided by applicant): Cancer is recognized as a highly complex disease involving myriad molecular processes and arises as the result of gradual accumulation of multiple genetic and proteomic alterations, which also serve as cancer biomarkers. A major focus of current cancer research is how to correlate these underlying molecular events with cancer development and progression. Recent advances in cancer molecular analysis and bioanalytical sciences have led to the development of DNA chips, ELISA, miniaturized biosensors, microfluidic devices (e.g., bioMEMS or microelectromechanical systems, and mass spectrometry. These enabling technologies have substantially influenced the way that we detect and analyze cancer, such as gene expression profiling, drug discovery, and clinical diagnostics. However, none of these technical platforms are sufficiently flexible to allow detection of both genetic alterations and protein profiles with sensitivity down to single molecule level. As current research in genomics and proteomics produces more sequence data, there is a strong need for new technologies that can rapidly screen a large number of nucleic acids and proteins. In this context, the primary goal of this proposal is to develop a versatile and sensitive technology that can quickly analyze cancer molecular profiles (such as DNAs, RNAs and proteins) in a highly multiplexed manner for accurate diagnostics, prognostics and effective therapeutics. The innovation and basic rationale of this technology lies in the novel optical properties of semiconductor quantum dots or QDs (e.g., tunable light emission, improved signal brightness, resistance against photobleaching, and simultaneous excitation of multiple colors) and our ability to make optical barcodes using these nanoparticles. Different from single QD based imaging; we propose to prepare QD encoded optical barcodes of both micrometer sizes and nanometer sizes. The micro-barcodes will be used to tag biomolecular probes, whereas the nanospheres will be used as reporters to enhance the current detection sensitivity by 2-3 orders of magnitude. We will also explore new surface encapsulation and pore sealing approaches to stabilize the optical barcodes, and chemical conjugation approaches to optimize biomolecular probe immobilization. We will further carry out experiments to detect mutations on both DNA and RNA level as well as the profiles of protein cancer biomarkers, which can be isolated from serum or homogenized cell and tissue specimens. We describe a new generation of QD-based optical barcoding technology for molecular analysis of cancer. The innovation arises from three levels: (1) the concept of using QDs for intensity-color based multiplexing, which has significantly higher multiplexing capability than traditional technology and simplified detection instrumentation; (2) the technique of using mesoporous material for micro barcodes and block-copolymer self assembly for nanobeads; and (3) the selection of clinically important biomarkers for cancer molecular analysis. It is built on our considerable expertise and strength in QD probe chemistry, optical imaging and cancer research. The major advantage of this versatile platform is that it allows simultaneous analysis of DNA, RNA as well as protein cancer markers with unprecedented sensitivity, which is not possible with other approaches.

描述（由申请人提供）：癌症是一种高度复杂的疾病，涉及无数的分子过程，是多种遗传和蛋白质组学改变逐渐积累的结果，这些改变也可以作为癌症的生物标志物。当前癌症研究的一个主要焦点是如何将这些潜在的分子事件与癌症的发生和进展联系起来。癌症分子分析和生物分析科学的最新进展导致了DNA芯片，ELISA，小型化生物传感器，微流体装置（例如，bioMEMS或微机电系统）和质谱的发展。这些使能技术极大地影响了我们检测和分析癌症的方式，如基因表达谱、药物发现和临床诊断。然而，这些技术平台都没有足够的灵活性来检测基因改变和蛋白质谱，灵敏度低至单分子水平。随着目前基因组学和蛋白质组学的研究产生了更多的序列数据，迫切需要能够快速筛选大量核酸和蛋白质的新技术。在此背景下，本提案的主要目标是开发一种多功能和敏感的技术，能够以高度多路复用的方式快速分析癌症分子谱（如dna， rna和蛋白质），以实现准确的诊断，预后和有效的治疗。这项技术的创新和基本原理在于半导体量子点或量子点的新型光学特性（例如，可调谐光发射，提高信号亮度，抗光漂白，同时激发多种颜色）以及我们使用这些纳米粒子制造光学条形码的能力。不同于单量子点成像；我们建议制备微米尺寸和纳米尺寸的量子点编码光学条形码。微条形码将用于标记生物分子探针，而纳米球将用作报告器，将目前的检测灵敏度提高2-3个数量级。我们还将探索新的表面封装和孔密封方法来稳定光学条形码，以及化学偶联方法来优化生物分子探针的固定。我们将进一步开展实验，检测DNA和RNA水平的突变，以及蛋白质癌症生物标志物的谱，这些生物标志物可以从血清或均质细胞和组织标本中分离出来。我们描述了用于癌症分子分析的新一代基于量子点的光学条形码技术。创新源于三个层面：(1)使用量子点进行基于强度颜色的多路复用的概念，与传统技术和简化的检测仪器相比，该概念具有明显更高的多路复用能力；(2)利用介孔材料制备微条形码和嵌段共聚物自组装纳米球的技术；(3)筛选临床重要的肿瘤分子分析生物标志物。它建立在我们在量子点探针化学，光学成像和癌症研究方面的专业知识和实力之上。这种多功能平台的主要优点是它允许同时分析DNA， RNA和蛋白质癌症标志物，具有前所未有的灵敏度，这是其他方法无法做到的。