How do mammalian cells handle mRNA therapeutics: Optimising the molecular basis of manufacture

利用信使核糖核酸产生针对新冠肺炎的疫苗，巩固了核酸开发预防病毒感染的方法并在威胁生命的癌症治疗中有更广泛应用的潜力(自然[2021]589：189-191；柳叶刀[2022]23：e450-e458)。为了最大限度地发挥mRNA(和其他基于核酸的疗法)的潜力，我们需要更多地了解它们进入细胞的传递、细胞内的处理/稳定性以及将细胞定位与有效表达联系起来的过程。我们还需要确定这些发现是否适用于细胞类型，或者过程是否可以针对特定的细胞类型和状态进行优化。这为您的项目提供了总体目标。您项目的具体目标将需要您(I)生成一组mRNA产品，(Ii)检查一组培养的哺乳动物细胞内的摄取、加工和表达，(Iii)根据产品表达的程度筛选宿主细胞的表型，(Iv)使用机器学习方法来了解哺乳动物细胞摄取mRNA产品后代谢途径的变化，以及(V)测试通过工程设计特定细胞功能得出的假设。为了实现这些目标，您将掌握广泛的分析、细胞和分子技术，包括荧光显微镜、高分辨率质谱分析、组学数据查询、化学计量学方法、数据解释以及细胞和过程工程。与过程创新中心(CPI)核糖核酸卓越中心(https://www.uk-cpi.com/about/national-centres/rna-centre-of-excellence)合作，我们将成为创新设计和制造核糖核酸疫苗和疗法的更广泛愿景的一部分。

The use of mRNA to generate vaccines against COVID-19 has cemented the potential for nucleic acids to develop approaches to prevent viral infection and to have wider applications in the treatment of life-threatening cancers (Nature [2021] 589: 189-191; The Lancet [2022] 23: e450-e458). To maximise the potential of mRNA (and other nucleic acid-based therapeutics) we need to understand more about their delivery into cells, intracellular handling/stability and the processes that link cellular localisation to effective expression. We also need to determine if findings are generic to cell types or if processes can be optimised for specific cell types and status. This provides the overall aim for your project. The specific objectives of your project will require you to (i) generate a panel of mRNA products (ii) examine the uptake, processing and expression within a panel of cultured mammalian cells, (iii) screen host cell phenotype in response to the extent of product expression, (iv) use machine learning methods to understand changes in metabolic pathways upon uptake of mRNA products by mammalian cells and (v) test hypotheses derived by engineering specific cellular functionalities. In achieving these objectives you will master a wide range of analytical, cellular and molecular techniques, including fluorescence microscopy, high resolution mass spectrometry assays, omics data interrogation, chemometrics methods, data interpretation and cellular and process engineering. Working with the Centre for Process Innovation (CPI) RNA Centre of Excellence (https://www.uk-cpi.com/about/national-centres/rna-centre-of-excellence) we will be part of a wider vision to innovate design and manufacture of RNA vaccines and therapeutics.