Optimising Product Quality of In Vitro Transcribed mRNA Therapeutics

基因疗法为治疗许多目前无法治愈的疾病提供了治疗希望，包括癌症、神经疾病和心血管疾病。然而，这些疗法缺乏支持其他生物制药开发的使能技术，导致进入临床的速度慢和/或产品性能差。例如，目前以信使核糖核酸为基础的基因疗法的信使核糖核酸的生产受到产量低和产品质量差的阻碍。这些mRNA分子是由体外转录(IVT)系统产生的，该系统包括四个生物输入组件：DNA模板、RNA聚合酶、核糖核苷三磷酸盐和帽类似物/帽酶。这些生物部分的功能和相互作用决定了‘正确’的mRNA产物和与不希望的产物相关的杂质的形成速度，例如无封顶的mRNA、长/短的dsRNA和长/短的单链RNA分子。这些杂质通过阻碍信使核糖核酸的性能(例如，每信使核糖核酸剂量体内产生的蛋白质)和激活有效的、不受欢迎的免疫反应来显著影响产品质量。因此，这些分子的不可预测和不可控制的形成是成功将IVT mRNA疗法转化为临床的关键障碍。该项目将为IVT mRNAs的生产创造新的生物部件，这些部件专门设计用于消除与产品相关的杂质的形成，同时也最大化系统产量和批次之间的重复性。

Gene therapy offers therapeutic promise for treating many currently incurable conditions, including cancers, neurological disorders and cardiovascular diseases. However, these therapies lack the enabling technologies that support development of other biopharmaceuticals, resulting in slow speed-to-clinic and/or poor product performance. For example, the production of mRNA for mRNA-based gene therapies is currently hampered by low yields and poor product qualities. These mRNA molecules are produced by in vitro transcription (IVT) systems that comprise four biological input components: the DNA template, an RNA polymerase, ribonucleoside triphosphates and a cap analogue/capping enzyme. The functionality of, and interplay between, these biological parts determines the rate of formation of both the 'correct' mRNA product, and undesirable product-related impurities, such as uncapped mRNA, long/short dsRNA, and long/short ssRNA molecules. These impurities significantly impact product quality, both by hampering mRNA performance (e.g. protein produced in vivo per mRNA dose) and activating potent, undesired immune responses. The unpredictable and uncontrollable formation of these molecules is therefore a critical obstacle to successfully translating IVT mRNA therapeutics into the clinic.This project will create new biological parts for production of IVT mRNAs, that are specifically designed to eliminate formation of product-related impurities, while also maximizing both system yield and batch-to-batch reproducibility.