Managing the sequence-specificity of antisense oligonucleotides in drug discovery.
Managing the sequence-specificity of antisense oligonucleotides in drug discovery.
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DOI：
10.1093/nar/gkx056
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发表时间：
2017-03-17
影响因子：
14.9
通讯作者：
Lindow M
中科院分区：
文献类型：
--
作者：
Hagedorn PH;Hansen BR;Koch T;Lindow M
所有的药物都会干扰接触到它们的细胞中许多基因的表达。这些基因表达变化可以分为由接合预期靶标引起的效应和由接合非预期靶标引起的效应。对于反义寡核苷酸，生物信息学算法的发展和序列数据库的质量，允许寡核苷酸序列进行计算分析，在其与预期和非预期的RNA靶标的相互作用的可预测性方面。应用这些工具能够选择序列特异性寡核苷酸，其中预期没有或只有很少的非预期RNA靶标。为了通过实验评估寡核苷酸序列特异性，我们推荐一种转录组学方案，其中一起评估两种或更多种靶向相同RNA分子但具有完全不同序列的寡核苷酸。这有助于澄清细胞RNA水平的哪些变化是由接合预期靶点的下游过程引起的，哪些可能与接合非预期靶点有关。根据所有类别药物的要求，在临床试验之前，必须在细胞和动物模型中评估寡核苷酸的潜在毒性。由于与非预期靶向相关的潜在不良反应是序列依赖性的，因此具有物种特异性，因此人细胞中的体外毒理学测定在寡核苷酸药物发现中尤其相关。
All drugs perturb the expression of many genes in the cells that are exposed to them. These gene expression changes can be divided into effects resulting from engaging the intended target and effects resulting from engaging unintended targets. For antisense oligonucleotides, developments in bioinformatics algorithms, and the quality of sequence databases, allow oligonucleotide sequences to be analyzed computationally, in terms of the predictability of their interactions with intended and unintended RNA targets. Applying these tools enables selection of sequence-specific oligonucleotides where no- or only few unintended RNA targets are expected. To evaluate oligonucleotide sequence-specificity experimentally, we recommend a transcriptomics protocol where two or more oligonucleotides targeting the same RNA molecule, but with entirely different sequences, are evaluated together. This helps to clarify which changes in cellular RNA levels result from downstream processes of engaging the intended target, and which are likely to be related to engaging unintended targets. As required for all classes of drugs, the toxic potential of oligonucleotides must be evaluated in cell- and animal models before clinical testing. Since potential adverse effects related to unintended targeting are sequence-dependent and therefore species-specific, in vitro toxicology assays in human cells are especially relevant in oligonucleotide drug discovery.