eprintid: 10180184
rev_number: 7
eprint_status: archive
userid: 699
dir: disk0/10/18/01/84
datestamp: 2023-11-06 17:38:11
lastmod: 2023-11-06 17:38:11
status_changed: 2023-11-06 17:38:11
type: article
metadata_visibility: show
sword_depositor: 699
creators_name: Woodruff, Rosie
creators_name: Parekh, Farhaan
creators_name: Lamb, Katarina
creators_name: Mekkaoui, Leila
creators_name: Allen, Christopher
creators_name: Smetanova, Katerina
creators_name: Huang, Jasmine
creators_name: Williams, Alex
creators_name: Toledo, Gerardo Santiago
creators_name: Lilova, Koki
creators_name: Roddie, Claire
creators_name: Sillibourne, James
creators_name: Pule, Martin
title: Large-scale manufacturing of base-edited chimeric antigen receptor T cells
ispublished: pub
divisions: UCL
divisions: B02
divisions: C10
divisions: D19
divisions: G98
keywords: base editing, BE4max, chimeric antigen receptor, circular RNAPD-1TIM3
note: © 2023 The Authors. Published by Elsevier under a Creative Commons license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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 × 108 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 × 106 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.
date: 2023-12-14
date_type: published
publisher: Elsevier BV
official_url: https://doi.org/10.1016/j.omtm.2023.101123
oa_status: green
full_text_type: pub
language: eng
primo: open
primo_central: open_green
verified: verified_manual
elements_id: 2103693
doi: 10.1016/j.omtm.2023.101123
medium: Electronic-eCollection
pii: S2329-0501(23)00162-6
lyricists_name: Pule, Martin
lyricists_id: MPULE38
actors_name: Flynn, Bernadette
actors_id: BFFLY94
actors_role: owner
full_text_status: public
publication: Molecular Therapy: Methods & Clinical Development
volume: 31
article_number: 101123
event_location: United States
citation:        Woodruff, Rosie;    Parekh, Farhaan;    Lamb, Katarina;    Mekkaoui, Leila;    Allen, Christopher;    Smetanova, Katerina;    Huang, Jasmine;                         ... Pule, Martin; + view all <#>        Woodruff, Rosie;  Parekh, Farhaan;  Lamb, Katarina;  Mekkaoui, Leila;  Allen, Christopher;  Smetanova, Katerina;  Huang, Jasmine;  Williams, Alex;  Toledo, Gerardo Santiago;  Lilova, Koki;  Roddie, Claire;  Sillibourne, James;  Pule, Martin;   - view fewer <#>    (2023)    Large-scale manufacturing of base-edited chimeric antigen receptor T cells.                   Molecular Therapy: Methods & Clinical Development , 31     , Article 101123.  10.1016/j.omtm.2023.101123 <https://doi.org/10.1016/j.omtm.2023.101123>.       Green open access   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/10180184/1/1-s2.0-S2329050123001626-main.pdf