21EBTA Novel Engineering Biology Solutions to the Production of Lentiviral Vectors by In vitro Assembly for Gene Therapy

细胞和基因治疗对患者有很大的好处，涵盖了广泛的严重疾病。根据细胞和基因治疗Catapult临床试验数据库，仅在英国，此类先进治疗药品（ATMP）的临床试验每年继续增加：2019年增加约45%，2020年比去年增加20%。据Catapult称，这占全球所有ATMP临床试验的12%，使英国成为该生物医学领域的全球领导者。这些先进的疗法需要载体来传递可以帮助对抗这些疾病的感兴趣的基因。最具商业前景的基因递送系统之一是慢病毒载体（LV）。在Catapult数据库中，LV最常用于体外治疗，尽管也报告了体内治疗的早期研究。LV携带的基因相对较大，并且具有假型，即，用包裹病毒颗粒表面的蛋白质来修饰LV，LV可以被设计成进入更广泛的细胞并传递感兴趣的基因。这些基因载体的供应（即，然而，在细胞和基因疗法的发展中仍然是一个主要的瓶颈（英国政府报告：2016年先进疗法制造工作组报告）。为了实现细胞和基因治疗的广泛应用和容易获得，需要具有成本效益的基因治疗载体制造，2016年ATMP工作组报告建议投资于病毒载体制造能力和能力。为了解决商业化生产过程中的病毒载体供应问题，载体生产方法需要改变为工业化工艺，该工艺具有稳健性并始终提供高产基因载体。通过工程生物学改进慢病毒载体生产有巨大的动机。这可以通过将功能载体的不同主要部分（病毒颗粒和包膜蛋白）结合在一起来构建LV来实现。这些技术的建立，以创建和验证这种新的LV生产系统模式，可以提高生物加工过程中的回收率。拟议的研究，建立在工程生物学的LV的最新发展，旨在创建和验证这些新的LV的生产，回收和纯化。具有成本效益的，简化的LV制造平台和我们在这项工作中开发的相关技术将有助于提高生物处理效率。因此，产品良率的提高最终将有助于满足低压电源的需求。这将增加基因载体的可用性，使基因和疗法更容易进入市场。这对许多患者来说都有巨大的潜在好处。更广泛地说，LV还广泛用于基因和细胞治疗领域之外，从基础生物学到药物筛选。我们在LV技术方面的创新将支持广泛的研究和开发，因此通常有助于生物科学和技术创新。

Cell and gene therapy promises great benefits to patients covering a wide range of serious diseases. In the UK alone, clinical trials for such advanced therapy medicinal products (ATMP) continue to rise annually according to the Cell and Gene Therapy Catapult clinical trials database: in 2019 the increase was about 45% and in 2020 the increase was 20% from previous year. According to the Catapult, this represents about 12% of all ATMP clinical trials globally making the UK a global leader in this field of biomedicine. These advanced therapies require carriers to deliver the genes of interest that can help in combatting these diseases. One of the most commercially promising gene delivery systems are lentiviral vectors (LVs). In the Catapult database, LVs are most commonly used for ex-vivo therapies although early phase studies are also reported for in-vivo therapies. The genes that LVs carry are relatively large and with pseudotyping, i.e., dressing up the LVs with proteins that envelope the surface of the viral particles, LVs can be designed to enter a broader range of cells and deliver the gene of interest. The supply of these gene vectors (i.e., gene carriers like LVs), however remain a major bottleneck in the development of cell and gene therapies (UK Government Report: 2016 Advanced Therapies Manufacturing Taskforce Report). Cost-efficient gene therapy vector manufacture is needed to realise wide applications of and easy access to cell and gene therapy with recommendations from the 2016 ATMP Taskforce Report to invest in viral vector manufacturing capacity and capability. To address viral vector supply issues during commercial manufacture, vector production methods need to change to an industrialised process that is robust and consistently delivers high yielding gene vectors. There is a huge incentive to improve lentiviral vector production through engineering biology. This can be achieved via novel construction of LVs by bringing together the different major parts of functional vector: the viral particle and the envelope proteins. The establishment of these techniques to create and characterise this new mode of LV production system could improve recovery yields during bioprocessing. The proposed research, which builds on recent developments in engineering biology of LVs, aims to create and characterise the production, recovery, and purification of these novel LVs.The cost-effective, simplified platforms for LV manufacture and associated technologies we develop in this work will help improve bioprocessing efficiencies. Therefore, the increase in product yield will eventually help to meet demand for LV supply. This will increase availability of gene vectors and make gene and therapies easier to reach the market. This has a huge potential benefit for many patients. More broadly, LVs are also widely used outside gene and cell therapy field from basic biology to drug screening. Our innovations in LV technology will support a wide range of research and development, therefore generally contribute to biological science and technology innovations.