Development of a stable, continuous lentiviral (LV) production system for improved gene delivery and cell function

开发稳定、连续的慢病毒 (LV) 生产系统，以改善基因传递和细胞功能

• 批准号: 2298917
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2298917
• 关键词: Development; stable; continuous; lentiviral; LV

Scalable and consistent production of stable cell lines for improved lentiviral (LV) vector production is necessary to ensure sufficient supply of high-quality vector for gene therapies and gene-modified cell therapies. This requires an integrated approach with both biological engineering of the cell lines and bioprocess engineering of the upstream and downstream processing being considered in parallel. From a biological engineering perspective, the project will seek to develop viral vectors which enable the simultaneous transduction and stimulation of T-cells (LentiStim) without the need for Dynabeads or antibodies in addition to undertaking a systematic, comparative study of various viral envelopes for enhanced T-cell engineering. From a bioprocessing perspective, the project will seek to establish a scalable bioprocess for the production of LentiStim viral vectors with the optimal viral envelope and develop an end-to-end process whereby the produced LV will be used to transduce primary T-cells with a chimeric antigen receptor (CAR). The T-cells produced from this process will be tested for anti-tumour efficacy in humanized mouse models. This will be established using NOD/SCID/IL2rynull (NSG) mouse model resistant to graft versus host disease with a B cell lymphoma cell line (Raji) or/and primary lymphoma cells.

为改进慢病毒(LV)载体的生产，需要可扩展和一致的稳定细胞系生产，以确保为基因治疗和基因修饰细胞治疗提供足够的高质量载体。这需要一种综合的方法，同时考虑细胞系的生物工程和上游和下游加工的生物工艺工程。从生物工程的角度来看，该项目将寻求开发能够同时转导和刺激T细胞(LentiSTim)的病毒载体，而不需要动力抗体或抗体，此外还将对各种病毒包膜进行系统的比较研究，以增强T细胞工程。从生物处理的角度来看，该项目将寻求建立一种可扩展的生物工艺，用于生产具有最佳病毒包膜的LentiSTim病毒载体，并开发一种端到端工艺，通过该工艺生产的LV将用于转导具有嵌合抗原受体(CAR)的原代T细胞。通过这一过程产生的T细胞将在人源化的小鼠模型中进行抗肿瘤效果测试。这将利用NOD/SCID/IL2rynull(NSG)小鼠模型建立抵抗移植物抗宿主病的B细胞淋巴瘤细胞系(Raji)或/和原代淋巴瘤细胞。