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

细胞和基因治疗有望为覆盖范围广泛的严重疾病的患者带来巨大好处。仅在英国，根据细胞和基因治疗Catapult临床试验数据库，此类高级治疗药物(ATMP)的临床试验持续每年上升：2019年增长约45%，2020年比前一年增长20%。根据Catapult的数据，这约占全球所有ATMP临床试验的12%，使英国成为这一生物医学领域的全球领先者。这些先进的疗法要求携带者提供有助于对抗这些疾病的感兴趣的基因。慢病毒载体是最具商业前景的基因传递系统之一。在Catapult数据库中，LV最常用于体外治疗，尽管体内治疗的早期研究也有报道。LV携带的基因相对较大，并且带有伪分型，即用包裹在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目标基因。然而，这些基因载体(即像LV这样的基因载体)的供应仍然是细胞和基因疗法发展的主要瓶颈(英国政府报告：2016高级疗法制造工作组报告)。根据2016年ATMP工作组报告的建议，需要成本效益高的基因治疗载体制造，以实现细胞和基因治疗的广泛应用和轻松获得，以投资于病毒载体制造能力和能力。为了解决商业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的过程，即稳健并持续地提供高产基因载体。通过工程生物学来改善慢病毒载体的生产是一个巨大的动机。这可以通过新的LV结构来实现，通过将功能载体的不同主要部分：病毒颗粒和包膜蛋白聚集在一起。建立这些技术来创造和表征这种新的LV生产系统模式，可以提高生物处理过程中的回收率。这项拟议的研究建立在LV工程生物学的最新发展基础上，旨在创造和表征这些新型LV的生产、回收和纯化。我们在这项工作中开发的成本效益高、简化的LV制造平台和相关技术将有助于提高生物处理效率。因此，产品产量的提高最终将有助于满足低压供应的需求。这将增加基因载体的可用性，并使基因和疗法更容易进入市场。这对许多患者有巨大的潜在好处。更广泛地说，LV也被广泛应用于从基础生物学到药物筛选的基因和细胞治疗领域之外。我们在LV技术方面的创新将支持广泛的研究和开发，因此总体上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.