CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients
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 w...
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Nature Publishing Group
2016
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4730182/ |
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pubmed-47301822016-02-03 CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients Su, Shu Hu, Bian Shao, Jie Shen, Bin Du, Juan Du, Yinan Zhou, Jiankui Yu, Lixia Zhang, Lianru Chen, Fangjun Sha, Huizi Cheng, Lei Meng, Fanyan Zou, Zhengyun Huang, Xingxu Liu, Baorui Article 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 Publishing Group 2016-01-28 /pmc/articles/PMC4730182/ /pubmed/26818188 http://dx.doi.org/10.1038/srep20070 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Su, Shu Hu, Bian Shao, Jie Shen, Bin Du, Juan Du, Yinan Zhou, Jiankui Yu, Lixia Zhang, Lianru Chen, Fangjun Sha, Huizi Cheng, Lei Meng, Fanyan Zou, Zhengyun Huang, Xingxu Liu, Baorui |
| spellingShingle |
Su, Shu Hu, Bian Shao, Jie Shen, Bin Du, Juan Du, Yinan Zhou, Jiankui Yu, Lixia Zhang, Lianru Chen, Fangjun Sha, Huizi Cheng, Lei Meng, Fanyan Zou, Zhengyun Huang, Xingxu Liu, Baorui CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients |
| author_facet |
Su, Shu Hu, Bian Shao, Jie Shen, Bin Du, Juan Du, Yinan Zhou, Jiankui Yu, Lixia Zhang, Lianru Chen, Fangjun Sha, Huizi Cheng, Lei Meng, Fanyan Zou, Zhengyun Huang, Xingxu Liu, Baorui |
| author_sort |
Su, Shu |
| title |
CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients |
| title_short |
CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients |
| title_full |
CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients |
| title_fullStr |
CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients |
| title_full_unstemmed |
CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients |
| title_sort |
crispr-cas9 mediated efficient pd-1 disruption on human primary t cells from cancer patients |
| description |
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. |
| publisher |
Nature Publishing Group |
| publishDate |
2016 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4730182/ |
| _version_ |
1613529572443160576 |