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Enhancement of the viability of T cells electroporated with DNA via osmotic dampening of the DNA-sensing cGAS-STING pathway.
- Source :
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Nature biomedical engineering [Nat Biomed Eng] 2024 Feb; Vol. 8 (2), pp. 149-164. Date of Electronic Publication: 2023 Jul 27. - Publication Year :
- 2024
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Abstract
- Viral delivery of DNA for the targeted reprogramming of human T cells can lead to random genomic integration, and electroporation is inefficient and can be toxic. Here we show that electroporation-induced toxicity in primary human T cells is mediated by the cytosolic pathway cGAS-STING (cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) synthase-stimulator of interferon genes). We also show that an isotonic buffer, identified by screening electroporation conditions, that reduces cGAS-STING surveillance allowed for the production of chimaeric antigen receptor (CAR) T cells with up to 20-fold higher CAR T cell numbers than standard electroporation and with higher antitumour activity in vivo than lentivirally generated CAR T cells. The osmotic pressure of the electroporation buffer dampened cGAS-DNA interactions, affecting the production of the STING activator 2'3'-cGAMP. The buffer also led to superior efficiencies in the transfection of therapeutically relevant primary T cells and human haematopoietic stem cells. Our findings may facilitate the optimization of electroporation-mediated DNA delivery for the production of genome-engineered T cells.<br /> (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)
Details
- Language :
- English
- ISSN :
- 2157-846X
- Volume :
- 8
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Nature biomedical engineering
- Publication Type :
- Academic Journal
- Accession number :
- 37500747
- Full Text :
- https://doi.org/10.1038/s41551-023-01073-7