A protein-based method for the generation of allogeneic chimeric antigen receptor T-cells

一种基于蛋白质的同种异体嵌合抗原受体 T 细胞生成方法

• 批准号: MR/S037144/1
• 负责人: Martin Pule
• 金额: $ 223.2万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FS037144%2F1
• 关键词: protein; based; method; generation; allogeneic

Chimeric Antigen Receptor (CAR) T-cell therapy has recently been approved in the USA and Europe for the treatment of relapsed B-cell malignancies, with potential further application in a broad range of tumour types. Currently, most CAR T-cell products are generated as autologous bespoke products for each patient. This has several disadvantages: each patient must undergo leukapheresis; production failure rate is ~10%; product quality is not guaranteed; there is a 3-4 week lag time to treatment; and economies of manufacturing scale cannot be applied.A potential solution is allogeneic CAR T-cells from healthy donors, which could be manufactured in advance, quality-checked and cryopreserved for timely administration to any potential patient. However, unrelated donor CAR T-cells would cause severe or fatal graft-versus-host-disease (taGvHD) mediated by the T-cell receptor (TCR) of donor T-cells. To deliver allogeneic CAR T-cells, TCR signalling must be prevented.To date, allogeneic CAR T-cell strategies have used genome-editing to delete the TCR. Indeed, we were the first to describe clinical use of anti-CD19 CAR T-cells generated from allogeneic TALEN-edited cells. While some clinical efficacy has been seen, a complex multi-stage manufacturing process is required, with highly stringent depletion of TCR positive T-cells. This difficult manufacture obviates much of the advantage in terms of practicality / cost of goods for allogeneic cells.We have since developed an alternative platform for allogeneic CAR T-cell manufacturing without genome-editing. By co-expressing a TCR-specific single-chain variable fragment with a Golgi retention signal (TCR-KDEL) along with the CAR, TCR-negative CAR T-cells can be generated with a single viral transduction. Further, additional co-expression of a GMP compliant marker gene facilitates facile highly stringent selection of TCR-negative, CAR-positive cells in a single sorting step.With this grant, we propose to test the TCR-KDEL allogeneic CAR T platform in patients with advanced B-cell malignancies, using a next-generation CD19 CAR (CAT19) currently in several studies at our centre. While we do not expect TCR-KDEL usage to be limited to CD19 CAR, testing with CAT19 allows direct comparison with ongoing autologous CAR studies, and acts as a 'proof of concept' for potential future studies. We anticipate our approach could reduce cost and broaden access to CAR T-cell therapy.

嵌合抗原受体（CAR）T细胞疗法最近在美国和欧洲被批准用于治疗复发性B细胞恶性肿瘤，并可能进一步应用于广泛的肿瘤类型。目前，大多数CAR T细胞产品都是为每位患者生成的自体定制产品。这有几个缺点：每个患者都必须进行白细胞分离;生产失败率约为10%;产品质量得不到保证;治疗有3-4周的滞后时间;并且不能应用生产规模的经济性。一种潜在的解决方案是来自健康供体的同种异体CAR T细胞，其可以提前生产、质量检查和冷冻保存，以便及时施用给任何潜在的患者。然而，不相关的供体CAR T细胞会引起由供体T细胞的T细胞受体（TCR）介导的严重或致命的移植物抗宿主病（taGvHD）。为了递送同种异体CAR T细胞，必须阻止TCR信号传导。迄今为止，同种异体CAR T细胞策略已经使用基因组编辑来删除TCR。事实上，我们是第一个描述从同种异体TALEN编辑的细胞产生的抗CD 19 CAR T细胞的临床用途的人。虽然已经看到一些临床功效，但需要复杂的多阶段制造过程，其中TCR阳性T细胞的高度严格消耗。这种困难的制造排除了同种异体细胞在实用性/商品成本方面的大部分优势。此后，我们开发了一种无需基因组编辑的同种异体CAR T细胞制造替代平台。通过与CAR一起共表达具有高尔基体滞留信号的TCR特异性单链可变片段（TCR-KDEL）沿着，可以用单次病毒转导产生TCR阴性CAR T细胞。此外，GMP合规标记基因的额外共表达有助于在单个分选步骤中轻松高度严格地选择TCR阴性，CAR阳性细胞。有了这笔资金，我们计划在晚期B细胞恶性肿瘤患者中测试TCR-KDEL同种异体CAR T平台，使用下一代CD 19 CAR（CAT 19）目前在我们中心的几项研究中。虽然我们不希望TCR-KDEL的使用仅限于CD 19 CAR，但使用CAT 19进行的测试允许与正在进行的自体CAR研究进行直接比较，并作为未来潜在研究的“概念验证”。我们预计我们的方法可以降低成本并扩大CAR T细胞治疗的可及性。