Optimised construct design for in vitro and in vivo therapeutic production

用于体外和体内治疗生产的优化构建体设计

• 批准号: 2602493
• 金额: --
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2602493
• 关键词: Optimised; construct; design; vitro; vivo

In vivo delivery of therapies encoded on nucleic acids is becoming an alternative for rapid development of biologic drugs and vaccines. Nucleic acids can be delivered in viral (AAV) and non-viral forms such as plasmid DNA and mRNA. However, interactions between the therapeutic vectors and the cellular hosts can prove detrimental for the high expression levels required for efficacious effects. Here we will work closely with AstraZeneca to design and assess vector design rules to maximise sustained therapeutic production. Our aim is to understand how to successfully transfer on to industrially-relevant therapeutic systems engineering design considerations testing these in vitro and, possibly, in vivo.The lab work will involve using Golden Gate assembly for the generation of different design configurations, transfection of these into cells and assessment of construct performance (production level) and cellular response. A model-guided approach will be adopted and will include a metabolic flux analysis model to build a pareto optimality front linking vector expression to specific cell growth rate for each design. The results will be used as a measure of process scalability and manufacturability.

核酸编码疗法的体内递送正成为生物药物和疫苗快速发展的另一种选择。核酸可以以病毒（AAV）和非病毒形式（如质粒DNA和mRNA）传递。然而，治疗载体和细胞宿主之间的相互作用可能不利于有效效果所需的高表达水平。在这里，我们将与阿斯利康密切合作，设计和评估载体设计规则，以最大限度地提高持续的治疗生产。我们的目标是了解如何成功地转移到工业相关的治疗系统工程设计考虑，在体外和可能的体内测试这些。实验室工作将包括使用金门组装来生成不同的设计配置，将这些配置转染到细胞中，并评估构建性能（生产水平）和细胞反应。将采用模型指导的方法，并将包括代谢通量分析模型，以建立将每种设计的载体表达与特定细胞生长速率联系起来的帕累托最优性前沿。结果将被用作过程可扩展性和可制造性的度量。