Discovery of Selective Small Molecule Probes for pre-microRNAs

 描述（由申请人提供）：MicroRNA（miRNAs）包括一个大家族，由约21 - 23个核苷酸的小非编码RNA组成，它们是基因表达的关键转录后调节因子，通过沉默靶mRNA的翻译发挥作用。到目前为止，大约有1,000种预测的人类miRNA被认为控制着超过60%的蛋白质编码基因的活性。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发挥关键作用。miRNA表达的改变、上调或下调与癌症、肥胖症、糖尿病、病毒感染和自身免疫性疾病、炎症、神经退行性疾病和心血管疾病等有关。这些联系使得靶向miRNA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miRNA成熟的选择性小分子抑制剂，从而为治疗人类疾病的下一代miR靶向疗法提供基础。为此，我们开发了一种概念新颖的创新方法，用于测定RNA-小分子相互作用，该方法利用催化信号放大的能力以及点击的选择性和生物正交性。 化学.通过这种平台测定技术，我们称之为使用酶联点击化学测定或cat-ELCCA的催化测定，我们设计了一种可以高通量实施的方法，几乎没有错误的读出，并且对于所有miRNA都是通用的。我们建议进一步开发和应用cat-ELCCA，以发现可用作靶向疾病中miRNA的化学探针的pre-miRNA选择性配体。

 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.