Optimization of mRNA untranslated regions for improved expression of therapeutic mRNA
Optimization of mRNA untranslated regions for improved expression of therapeutic mRNA
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DOI：
10.1080/15476286.2018.1450054
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发表时间：
2018-01-01
期刊：
影响因子：
4.1
通讯作者：
Brown, Jeffrey M.
中科院分区：
文献类型：
--
作者：
Asrani, Kirtika H.;Farelli, Jeremiah D.;Brown, Jeffrey M.
基于mRNA的疗法对于治疗遗传疾病具有很大的希望。然而，这种治疗方法受到多种挑战，包括外源性施用的mRNA的短半衰期和随后的蛋白质产生。非翻译区（UTR）的调节代表了一种增强mRNA稳定性和翻译效率的方法。目前的研究描述并验证了使用一组不同的5UTR和3UTR组合的筛选方法，以改善精氨酸酶1（ARG1）蛋白（一种潜在的治疗mRNA靶点）的表达。数据揭示了在开发用于工程mRNA改进的筛选方法时需要考虑的许多关键方面。首先，基于质粒的筛选方法与由外源表达的mRNA驱动的蛋白质表达不相关。其次，ARG1蛋白产量的提高是由翻译增加而不是mRNA稳定性的提高驱动的。最后，5个UTR似乎是外源递送mRNA的蛋白质表达的关键驱动因素。根据组合文库的测试，补体因子3（C3）和细胞色素p4502E1（CYP2E1）的5UTR显示相对于参考UTR的蛋白质表达的最大和最一致的增加。总的来说，这些数据为治疗性mRNA的开发和优化提供了重要信息。
mRNA based therapies hold great promise for the treatment of genetic diseases. However, this therapeutic approach suffers from multiple challenges including the short half-life of exogenously administered mRNA and subsequent protein production. Modulation of untranslated regions (UTR) represents one approach to enhance both mRNA stability and translation efficiency. The current studies describe and validate screening methods using a diverse set of 5UTR and 3UTR combinations for improved expression of the Arginase 1 (ARG1) protein, a potential therapeutic mRNA target. Data revealed a number of critical aspects which need to be considered when developing a screening approach for engineering mRNA improvements. First, plasmid-based screening methods do not correlate with protein expression driven by exogenously expressed mRNA. Second, improved ARG1 protein production was driven by increased translation and not improved mRNA stability. Finally, the 5 UTR appears to be the key driver in protein expression for exogenously delivered mRNA. From the testing of the combinatorial library, the 5UTR for complement factor 3 (C3) and cytochrome p4502E1 (CYP2E1) showed the largest and most consistent increase in protein expression relative to a reference UTR. Collectively, these data provide important information for the development and optimization of therapeutic mRNAs.