Optical probe for the flow cytometric screening of microRNA

用于流式细胞术筛选 microRNA 的光学探针

• 批准号: 7186920
• 负责人: Andrew Tsourkas
• 金额: $ 17.16万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7186920
• 关键词: Biological Assay; Breast Cancer Cell; Cancer Detection; Cancer cell line; Cell Count; Cells; Chromosome Fragile Sites; Class; Clinical; Condition; Diagnosis; Diagnostic; Disruption; Dyes; Encapsulated; Event; Flow Cytometry; Functional RNA; Gene Expression; Genes; Genomics; Goals; HIV; Human; Hybrids; Individual; Length; Life; Link; Malignant Neoplasms; Measurement; Measures; Messenger RNA; Methods; MicroRNAs; Molecular; Normal Cell; Northern Blotting; Nucleotides; Numbers; Oligonucleotides; Optics; Pathogenesis; Peptides; Population; Property; Protocols documentation; Reporter; Reporting; Research; Screening for cancer; Screening procedure; Signal Transduction; Specimen; Standardization; Technology; Vertebral column; base; design; high throughput screening; imaging probe; molecular imaging; outcome forecast

DESCRIPTION (provided by applicant): It has recently been reported that over 52% of microRNA (miRNA) genes are in cancer-associated genomic regions or in fragile sites. Moreover, an increasing number of studies show that the deregulation of specific miRNAs are linked to different forms of cancer; suggesting that miRNA may have diagnostic potential. Current approaches to measuring miRNA levels, however, lack the sensitivity to detect a small number of cells with altered miRNA expression in the presence of a large number of "normal" cells in human specimens. The overall goal of this proposal is to develop a molecular imaging probe that will allow miRNA expression levels to be measured at the single living cell level via flow cytometry. Since miRNAs are shorter, more stable, and much more abundant than messenger RNAs within individual cells, our plans to target miRNA with 'quantitative molecular beacons' (QMBs) has a significant inherent advantage compared with mRNA-based methods. Moreover, the use of flow cytometry allows for the highly sensitive, high-throughput screening of single cells. Therefore, we hypothesize that the proposed QMB technology could provide an important advance in the early detection of cancer. The specific aims for this proposal are (1) optimize the optical properties and the delivery of QMBs into breast cancer cells and (2) evaluate the sensitivity, precision, and accuracy of the QMB-based approach in detecting aberrant miRNA expression at the single cell level. The long-term goal of this research is to further develop the QMB technology into a simple clinical assay for cancer detection, diagnosis, and prognosis.

描述（由申请人提供）：最近据报道，超过52％的microRNA（miRNA）基因在与癌症相关的基因组区域或脆弱部位中。此外，越来越多的研究表明，特定miRNA的放松管制与不同形式的癌症有关。表明miRNA可能具有诊断潜力。然而，当前测量miRNA水平的方法缺乏在人类标本中大量“正常”细胞存在下检测少数细胞具有改变miRNA表达的细胞的敏感性。该建议的总体目标是开发一种分子成像探针，该探针将通过流式细胞仪在单个活细胞水平上测量miRNA表达水平。由于miRNA比单个细胞中的Messenger RNA更短，更稳定，更丰富，因此与基于mRNA的方法相比，我们用“定量分子信标”（QMB）靶向miRNA的计划具有显着的固有优势。此外，流式细胞仪的使用允许对单个细胞进行高度敏感的高通量筛选。因此，我们假设所提出的QMB技术可以在早期发现癌症方面提供重要的进步。该建议的具体目的是（1）优化QMB的光学特性和递送到乳腺癌细胞中，（2）评估基于QMB的方法在检测单细胞水平上异常miRNA表达时的敏感性，精度和准确性。这项研究的长期目标是将QMB技术进一步发展为一种简单的临床检测，诊断和预后。