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）：癌症被认为是一种高度复杂的疾病，涉及多种分子过程，并且由于多种遗传和蛋白质组学改变的逐渐积累而产生，这也是癌症生物标志物。当前癌症研究的主要重点是如何将这些基本分子事件与癌症的发展和进展相关联。癌症分子分析和生物分析科学的最新进展已导致DNA芯片，ELISA，微型生物传感器，微流体装置（例如，生物学或微电力机械系统或微观机械系统和质谱仪和质谱仪的群量仪，这些启用技术都具有很大的表达和分析，例如基因癌症，例如，生物机电系统或微观力学系统或微观力学系统和质谱量。但是，随着目前的基因组学和蛋白质组学的研究，这些技术平台都没有足够的灵活性，可以降低遗传变化和蛋白质特征，以降低基因组学和蛋白质组学的研究。以高度多重的方式分析癌症分子谱（例如DNA，RNA和蛋白质），以进行准确的诊断，预后和有效的疗法。这项技术的创新和基本原理在于半导体量子点或QD的新型光学特性（例如，可调的光发射，改善的信号亮度，对光漂白的抵抗力以及对多种颜色的同时激发的电阻）以及我们使用这些NanAparticles制作光学条形码的能力。不同于基于QD的成像；我们建议准备QD编码的千分尺和纳米尺寸的光学条形码。微型键盘将用于标记生物分子探针，而纳米球将用作记者，以增强当前的检测灵敏度，降低2-3个数量级。我们还将探索新的表面封装和孔密封方法，以稳定光学条形码，以及化学共轭方法，以优化生物分子探针固定。我们将进一步进行实验，以检测DNA和RNA水平上的突变以及蛋白质癌生物标志物的特征，这些突变可以从血清或均质的细胞和组织标本中分离出来。 我们描述了一种新一代的基于QD的光条形码技术，用于癌症的分子分析。创新源于三个层次：（1）将QD用于基于强度色的多重功能的概念，其多路复用能力明显高于传统技术和简化的检测仪器； （2）将介孔材料用于微型条形码和纳米胶的块状聚合物自组装的技术； （3）选择临床上重要的生物标志物进行癌症分子分析。它建立在我们在QD探针化学，光学成像和癌症研究中的大量专业知识和力量上。这个多功能平台的主要优点是，它允许同时分析具有前所未有的灵敏度的DNA，RNA和蛋白质癌标记物，而其他方法则是不可能的。