[HTML][HTML] CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients

S Su, B Hu, J Shao, B Shen, J Du, Y Du, J Zhou, L Yu… - Scientific reports, 2016 - nature.com
S Su, B Hu, J Shao, B Shen, J Du, Y Du, J Zhou, L Yu, L Zhang, F Chen, H Sha, L Cheng…
Scientific reports, 2016nature.com
Strategies that enhance the function of T cells are critical for immunotherapy. One negative
regulator of T-cell activity is ligand PD-L1, which is expressed on dentritic cells (DCs) or
some tumor cells and functions through binding of programmed death-1 (PD-1) receptor on
activated T cells. Here we described for the first time a non-viral mediated approach to
reprogram primary human T cells by disruption of PD-1. We showed that the gene knockout
of PD-1 by electroporation of plasmids encoding sgRNA and Cas9 was technically feasible …
Abstract
Strategies that enhance the function of T cells are critical for immunotherapy. One negative regulator of T-cell activity is ligand PD-L1, which is expressed on dentritic cells (DCs) or some tumor cells and functions through binding of programmed death-1 (PD-1) receptor on activated T cells. Here we described for the first time a non-viral mediated approach to reprogram primary human T cells by disruption of PD-1. We showed that the gene knockout of PD-1 by electroporation of plasmids encoding sgRNA and Cas9 was technically feasible. The disruption of inhibitory checkpoint gene PD-1 resulted in significant reduction of PD-1 expression but didn’t affect the viability of primary human T cells during the prolonged in vitro culture. Cellular immune response of the gene modified T cells was characterized by up-regulated IFN-γ production and enhanced cytotoxicity. These results suggest that we have demonstrated an approach for efficient checkpoint inhibitor disruption in T cells, providing a new strategy for targeting checkpoint inhibitors, which could potentialy be useful to improve the efficacy of T-cell based adoptive therapies.
nature.com