Increased functional potency of multi-edited CAR-T cells manufactured by a non-viral transfection system

Increased functional potency of multi-edited CAR-T cells manufactured by a non-viral transfection system

Chimeric antigen receptor (CAR)-T cell therapy represents a breakthrough for the treatment of hematological malignancies. However, to treat solid tumors and certain hematologic cancers, next-generation CAR-T cells require further genetic modifications to overcome some of the current limitations. Imp...

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Bibliographic Details
Main Authors: Aine O’Sullivan, Sarah Case, Aisling McCrudden, Emer Hackett, Louise Gallagher, Darren Martin, Gillian P. Johnson, Kirti Mahadik, Thomas Kienzle, Jude Kevin Lim, Aya Nashat, Kartik Srinivasan, Mark W. Lowdell, Lisa O’Flynn, Jamie Frankish
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Molecular Therapy: Methods & Clinical Development
Subjects:
CAR-T
genetic engineering
adoptive cell therapy
metabolism
cell avidity
cell cytotoxicity
Online Access:http://www.sciencedirect.com/science/article/pii/S2329050124002055
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Summary:Chimeric antigen receptor (CAR)-T cell therapy represents a breakthrough for the treatment of hematological malignancies. However, to treat solid tumors and certain hematologic cancers, next-generation CAR-T cells require further genetic modifications to overcome some of the current limitations. Improving manufacturing processes to preserve cell health and function of edited T cells is equally critical. Here, we report that Solupore, a Good Manufacturing Practice-aligned, non-viral physicochemical transfection system, can be used to manufacture multi-edited CAR-T cells using CRISPR-Cas9 ribonucleoproteins while maintaining robust cell functionality. When compared to electroporation, an industry standard, T cells that were triple edited using Solupore had reduced levels of apoptosis and maintained similar proportions of stem cell memory T cells with higher oxidative phosphorylation levels. Following lentiviral transduction with a CD19 CAR, and subsequent cryopreservation, triple-edited CAR-T cells manufactured using Solupore demonstrated improved immunological synapse strength to target CD19+ Raji cells and enhanced cellular cytotoxicity compared with electroporated CAR-T cells. In an in vivo mouse model (NSG), Solupore triple-edited CAR-T cells enhanced tumor growth inhibition by more than 30-fold compared to electroporated cells.
ISSN:2329-0501

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