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Large-scale manufacturing of base-edited chimeric antigen receptor T cells.
- Source :
-
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). - Publication Year :
- 2023
-
Abstract
- 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 <superscript>8</superscript> 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 <superscript>6</superscript> 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.<br />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.<br /> (© 2023 The Authors.)
Details
- Language :
- English
- ISSN :
- 2329-0501
- Volume :
- 31
- Database :
- MEDLINE
- Journal :
- Molecular therapy. Methods & clinical development
- Publication Type :
- Academic Journal
- Accession number :
- 37886606
- Full Text :
- https://doi.org/10.1016/j.omtm.2023.101123