New Assays for Expression Profiling of MicroRNAs

MicroRNA 表达谱的新检测方法

• 批准号: 8902301
• 负责人: Weijun Liu
• 金额: $ 22.5万
• 依托单位: NAWGEN, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8902301
• 关键词: Address; Algorithms; Archives; Bioinformatics; Biological; Biological Assay; Biological Markers; Biological Process; Clinical; Code; Data; Databases; Development; Formalin; Gene Expression; Gene Targeting; Genes; Goals; Gold; High-Throughput RNA Sequencing; Human; Human Genome; Individual; Lead; Malignant Neoplasms; Methods; MicroRNAs; Molecular Profiling; Paraffin Embedding; Proteins; Publishing; RNA; RNA Sequences; Regulator Genes; Research; Research Infrastructure; Research Personnel; Reverse Transcription; Sampling; Services; Testing; Time; Tissue Sample; Tissues; Tumor Tissue; Validation; Work; base; biological research; cancer type; clinical application; design; human disease; miRNA expression profiling; novel; public health relevance; research study; success; transcriptome sequencing

 DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) are involved in many diverse biological processes and a single miRNA can modulate the expression of hundreds of gene targets. Changes in miRNA expression could have profound biological impacts, leading to a variety of human diseases. To date, most miRNA expression profiling studies have relied on microarrays or high-throughput RNA sequencing (RNA-seq). For these high-throughput experiments, it is important to validate the profiling results with independent methods. Real-time PCR is considered as the gold standard for gene expression quantification and thus is ideal for validation of microarray or RNA-seq results. Furthermore, despite their wide use for miRNA profiling, there are some major limitations associated with microarrays and RNA-seq, such as the requirement of high-quality RNA samples. To address these issues, we have developed a new real-time RT-PCR method. With this new method, we have profiled the expression of cancer-related miRNAs in hundreds of archived tumor tissues, and identified novel miRNA signatures as predictive biomarkers for multiple types of cancer. Building on the success of our new method for cancer-related miRNA profiling, we propose to significantly expand the scope of the method by designing and testing new assays to encompass all known miRNAs in the human genome (Specific Aim 1). These miRNA assays will serve two main purposes: 1) As a reliable independent method for validation of high-throughput results from microarrays or RNA-seq; and 2) expression profiling of low- quality degraded RNA extracted from clinical tissues. In both cases, availability of these miRNA assays will enable researchers to correlate abnormal miRNA expression changes to human diseases by focusing on a selected set of miRNAs. To facilitate the selection of miRNA assays by end users, we also propose to establish an online database to present experimentally validated assays for all known human miRNAs (Specific Aim 2).

 描述（由申请人提供）：微小RNA（miRNA）参与许多不同的生物过程，单个miRNA可以调节数百个基因靶点的表达。miRNA表达的变化可能产生深远的生物学影响，导致各种人类疾病。迄今为止，大多数miRNA表达谱研究都依赖于微阵列或高通量RNA测序（RNA-seq）。对于这些高通量实验，重要的是用独立的方法验证分析结果。实时PCR被认为是基因表达定量的金标准，因此是验证微阵列或RNA-seq结果的理想方法。此外，尽管它们广泛用于miRNA谱分析，但微阵列和RNA-seq存在一些与之相关的主要限制，例如需要高质量的RNA样品。为了解决这些问题，我们开发了一种新的实时RT-PCR方法。通过这种新方法，我们分析了数百种存档肿瘤组织中癌症相关miRNA的表达，并确定了新的miRNA特征作为多种类型癌症的预测生物标志物。基于我们成功的癌症相关miRNA分析新方法，我们建议通过设计和测试新的检测方法来显着扩大该方法的范围，以涵盖人类基因组中所有已知的miRNA（具体目标1）。这些miRNA测定将用于两个主要目的：1）作为用于验证来自微阵列或RNA-seq的高通量结果的可靠的独立方法;和2）从临床组织提取的低质量降解RNA的表达谱分析。在这两种情况下，这些miRNA检测的可用性将使研究人员能够通过关注一组选定的miRNA来将异常miRNA表达变化与人类疾病相关联。为了方便最终用户选择miRNA检测方法，我们还建议建立一个在线数据库，以提供所有已知人类miRNA的实验验证检测方法（特定目标2）。