Large-scale manufacturing of base-edited chimeric antigen receptor T cells.

Autor:	Woodruff R; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Parekh F; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Lamb K; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Mekkaoui L; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Allen C; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Smetanova K; Imperial College London, South Kensington Campus, SW7 2AZ London, UK., Huang J; Imperial College London, South Kensington Campus, SW7 2AZ London, UK., Williams A; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Toledo GS; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Lilova K; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Roddie C; Deparment of Haematology, Cancer Institute, 72 Huntley Street, WC1E 6BT London, UK., Sillibourne J; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK., Pule M; Autolus Therapeutics, The Mediaworks, 191 Wood Lane, W12 7FP London, UK.; Deparment of Haematology, Cancer Institute, 72 Huntley Street, WC1E 6BT London, UK.
Jazyk:	angličtina
Zdroj:	Molecular therapy. Methods & clinical development [Mol Ther Methods Clin Dev] 2023 Oct 05; Vol. 31, pp. 101123. Date of Electronic Publication: 2023 Oct 05 (Print Publication: 2023).
DOI:	10.1016/j.omtm.2023.101123
Abstrakt:	Base editing is a revolutionary gene-editing technique enabling the introduction of point mutations into the genome without generating detrimental DNA double-stranded breaks. Base-editing enzymes are commonly delivered in the form of modified linear messenger RNA (mRNA) that is costly to produce. Here, we address this problem by developing a simple protocol for manufacturing base-edited cells using circular RNA (circRNA), which is less expensive to synthesize. Compared with linear mRNA, higher editing efficiencies were achieved with circRNA, enabling an 8-fold reduction in the amount of RNA required. We used this protocol to manufacture a clinical dose (1 × 10 8 cells) of base-edited chimeric antigen receptor (CAR) T cells lacking expression of the inhibitory receptor, PD-1. Editing efficiencies of up to 86% were obtained using 0.25 μg circRNA/1 × 10 6 cells. Increased editing efficiencies with circRNA were attributed to more efficient translation. These results suggest that circRNA, which is less expensive to produce than linear mRNA, is a viable option for reducing the cost of manufacturing base-edited cells at scale. Competing Interests: R.W., F.P., K.L., L.M., C.A., A.W., G.S.T, K.L., J.S., and M.P. are employees of Autolus Therapeutics plc and hold equity shares or options in the company. M.P. receives royalty share from patents licensed to Autolus Therapeutics plc. (© 2023 The Authors.)
Databáze:	MEDLINE
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