Discovery of Selective Small Molecule Probes for pre-microRNAs

 描述(申请人提供)：microRNAs(MiRNAs)包括一个小的、~21个−的23个核苷酸的非编码RNA的大家族，它们已经成为基因表达的关键转录后调节因子，并通过沉默靶mRNAs的翻译而发挥作用。到目前为止，有大约1000个预测的人类miRNAs被认为控制着所有蛋白质编码基因的60%的活性。毫不奇怪，这些小RNA已被证明在从发育到疾病的几乎所有人类生物学方面都发挥着关键作用。MiRNA表达的改变，无论是上调还是下调，都与癌症、肥胖、糖尿病、病毒感染和自身免疫、炎症、神经退行性疾病和心血管疾病等有关。这些联系使得以miRNAs为靶点作为一种新的治疗策略具有吸引力。我们的总体目标是发现和开发选择性的miRNA成熟小分子抑制剂，从而为下一代miR靶向疗法治疗人类疾病提供基础。为了做到这一点，我们开发了一种全新的、创新的方法来分析rna-小分子相互作用，该方法利用了催化信号放大的能力以及Click的选择性和生物正交性。 化学反应。通过这种平台分析技术，我们称之为酶联点击化学分析或CAT-ELCCA的催化分析，我们设计了一种可以高通量实施的方法，几乎没有错误读出，并且对所有miRNAs都是通用的。我们建议进一步开发和应用CAT-ELCCA来发现前miRNA选择性配体，这些配体可以用作疾病中靶向miRNAs的化学探针。

 DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) comprise a large family of small, ~21−23 nucleotide noncoding RNAs that have emerged as key post-transcriptional regulators of gene expression and act by silencing the translation of target mRNAs. To date, there are ~1,000 predicted human miRNAs believed to control the activity of >60% of all protein-coding genes. Not surprisingly, these small RNAs have been shown to play crucial roles in nearly all aspects of human biology from development to disease. Alteration of miRNA expression, up- or downregulation, has been linked to cancer, obesity, diabetes, viral infections and autoimmune, inflammatory, neurodegenerative and cardiovascular diseases among others. These connections have made the targeting of miRNAs attractive as a novel therapeutic strategy. Our overall goal is to discover and develop selective small molecule inhibitors of miRNA maturation, thus providing the basis for next-generation miR-targeted therapeutics for the treatment of human disease. To do so, we have developed a conceptually new and innovative approach for assaying RNA-small molecule interactions that takes advantage of the power of catalytic signal amplification combined with the selectivity and bioorthogonality of click chemistry. Through this platform assay technology, which we term catalytic assay using enzyme-linked click chemistry assay or cat- ELCCA, we have designed a method that can be implemented in high-throughput, is virtually free of false read- outs and is general for all miRNAs. We propose to further develop and apply cat-ELCCA toward the discovery of pre-miRNA-selective ligands that can be used as chemical probes for targeting miRNAs in disease.