ADAPTING PADLOCK-PROBE DETECTION SYSTEMS TO NCRNA DETECTION FOR CANCER DIAGNOSTI

将挂锁探针检测系​​统应用于 NCRNA 检测以进行癌症诊断

• 批准号: 8340289
• 负责人: JOHN COOPER
• 金额: $ 19.87万
• 依托单位: CUSTOMARRAY, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-23 至 2012-06-22
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8340289
• 关键词: Algorithms; Appearance; Biological Assay; Biological Markers; Cancer Detection; Cancer Diagnostics; Cancer Patient; Colon Carcinoma; Computer software; DNA Microarray Chip; Data; Data Analyses; Data Set; Detection; Development; Functional RNA; Genetic Transcription; Human; Malignant Neoplasms; Malignant neoplasm of ovary; Malignant neoplasm of prostate; MicroRNAs; Nucleotides; Oligonucleotides; Patients; Pattern; Phase; Proliferating; Protocols documentation; Sampling; Serum; Signal Transduction; Specificity; Structure; System; Testing; Work; base; cancer type; extracellular; neoplastic cell; novel

This proposal seeks to develop a cancer diagnostic based on microRNA (miRNA) expression patterns found in human serum. Mounting evidence is showing that patterns of miRNA expression and the appearance of these signals in circulating extra-cellular compartments are hallmarks of proliferating tumor cells. Our preliminary results have shown a clear ability to distinguish sera derived from cancer patients for 5 different types of cancer. In a data set involving 246 patient-derived serum samples, we were able to distinguish cancer derived sera from normal controls with a sensitivity of 76% and a specificity of 84%. These data, though promising, also indicated to us that a more sensitive assay is necessary in order to resolve these distinct patterns of miRNA expression. To this end, we propose the development of a multiplexed padlock probe assay, integrated with microarray detection that will simultaneously detect most of the known human miRNAs. Our unique customizable DNA microarray synthesis platform allows us to both produce the padlock probes and to detect signal from the amplified mixture. Finally, using this assay, we will investigate the extracellular compartment or structure that protects the miRNA molecules that are secreted by proliferating tumor cells. In phase II, we will apply this assay to look for patterns of miRNA and other non-coding RNA (ncRNA) expression from normal and cancer patient serum samples, repeating our previous work with this novel assay.

这项建议寻求开发一种基于人血清中发现的microRNA(MiRNA)表达模式的癌症诊断方法。越来越多的证据表明，miRNA的表达模式和这些信号出现在循环的细胞外隔间是肿瘤细胞增殖的标志。我们的初步结果表明，我们有明显的能力区分5种不同类型的癌症患者的血清。在一个涉及246个患者血清样本的数据集中，我们能够区分癌症患者血清和正常对照，敏感性为76%，特异性为84%。这些数据虽然很有希望，但也向我们表明，为了解决这些不同的miRNA表达模式，有必要进行更灵敏的分析。为此，我们提出了一种多重挂锁探针分析的发展，与微阵列检测相结合，将同时检测大多数已知的人类miRNAs。我们独特的可定制DNA微阵列合成平台使我们既可以生产挂锁探针，也可以从放大的混合物中检测信号。最后，利用这个实验，我们将研究保护由增殖的肿瘤细胞分泌的miRNA分子的细胞外间隔或结构。在第二阶段，我们将应用这项检测来寻找正常和癌症患者血清样本中miRNA和其他非编码RNA(NcRNA)的表达模式，重复我们之前使用这一新检测方法所做的工作。