In Vitro Validation of miRNA-Mediated Gene Expression Linked to Drug Metabolism.
In Vitro Validation of miRNA-Mediated Gene Expression Linked to Drug Metabolism.
复制标题
与药物代谢相关的miRNA介导的基因表达的体外验证。
DOI：
10.1002/cpph.30
复制
发表时间：
2017-12-20
影响因子：
--
通讯作者：
Ajit SK
中科院分区：
文献类型：
--
作者：
Shenoda BB;Ramanathan S;Ajit SK
用于研究基因如何影响药物反应的药物基因组学方法对于设计旨在最大化疗效和最小化不良反应的个性化治疗至关重要。药物功效通常取决于药物靶基因或参与治疗剂代谢和转运的基因的序列和表达水平。反过来，这些基因的表达受到小的非编码miRNA的负调控。体液中的miRNA水平已被广泛研究作为潜在的诊断和预后生物标志物。研究表明，miRNA调控多个基因，序列同源性用于预测哪些基因受到特定miRNA的调控。一旦基因被鉴定为目标miRNA的潜在靶标，则进行实验以确认miRNA与靶基因相互作用并且可以改变其表达水平和/或其活性。例如，在治疗复杂区域疼痛综合征之前和之后，已经报道了从对氯胺酮的良好和不良反应者获得的全血中miRNA的差异表达。在这种情况下，hsa-miR-548 d-5 p在不良反应者中相对于良好反应者显著较低。该miRNA被预测为靶向UDP-葡萄糖醛酸基转移酶1A 1（UGT 1A 1），一种关键的药物代谢酶。本单元描述了用于确认miR-548 d-5 p介导的UGT 1A 1调控的方案。所描述的方法可以广泛地用于验证任何基因的miRNA介导的负调控。确定影响药物代谢的酶和转运蛋白的miRNA介导的调节是设计个性化治疗和理解对治疗剂的反应变化的机制的关键步骤。
Pharmacogenomic approaches used to investigate how genes affect drug responses are critical for designing personalized therapies aimed at maximizing efficacy and minimizing adverse effects. Drug efficacy is often dependent on the sequence and expression levels of drug target genes or those involved in the metabolism and transport of the therapeutic agent. Expression of these genes, in turn, is negatively regulated by small noncoding miRNAs. The levels of miRNAs in bodily fluids have been studied extensively as potential diagnostic and prognostic biomarkers. Studies have shown that miRNAs regulate multiple genes and sequence homology is used to predict which genes are subject to regulation by a particular miRNA. Once a gene is identified as a potential target for an miRNA of interest, experiments are undertaken to confirm that the miRNA interacts with the target gene and can alter its level of expression and/or its activity. For example, the differential expression of miRNAs in whole blood obtained from good and poor responders to ketamine has been reported both prior to, and following treatment for complex regional pain syndrome. In this case, hsa-miR-548d-5p was significantly lower in poor responders relative to good responders. This miRNA was predicted to target UDP-glucuronyl transferase 1A1 (UGT1A1), a key drug metabolizing enzyme. Described in this unit are protocols used to confirm miR-548d-5p-mediated UGT1A1 regulation. The approaches described can be employed broadly for the validation of miRNA-mediated negative regulation of any gene. Determining miRNA-mediated regulation of enzymes and transporters affecting drug metabolism is a critical step in designing personalized therapy and for understanding the mechanisms responsible for variations in the responses to therapeutic agents.