PMID- 36532006 OWN - NLM STAT- MEDLINE DCOM- 20221220 LR - 20231213 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 13 DP - 2022 TI - Simultaneous editing of TCR, HLA-I/II and HLA-E resulted in enhanced universal CAR-T resistance to allo-rejection. PG - 1052717 LID - 10.3389/fimmu.2022.1052717 [doi] LID - 1052717 AB - INTRODUCTION: The major challenge for universal chimeric antigen receptor T cell (UCAR-T) therapy is the inability to persist for a long time in patients leading to inferior efficacy clinically. The objective of this study was to design a novel UCAR-T cell that could avoid the occurrence of allo-rejection and provide effective resistance to allogeneic Natural Killer (NK) cell rejection, together with the validation of its safety and efficacy ex vivo and in vivo. METHODS: We prepared T-cell receptor (TCR), Human leukocyte antigen (HLA)-I/II triple-edited (TUCAR-T) cells and evaluated the anti-tumor efficacy ex vivo and in vivo. We measured the resistance of exogenous HLA-E expressing TUCAR-T (ETUCAR-T) to NK rejection by using an enhanced NK. Furthermore, we established the safety and efficacy of this regimen by treating Nalm6 tumor-bearing mice with a repeated high-dose infusion of ETUCAR-T. Moreover, we analyzed the effects of individual gene deficiency CAR-T on treated mice and the changes in the transcriptional profiles of different gene-edited T cells via RNA-Seq. RESULTS: Data showed that HLA-II editing didn't impair the anti-tumor efficacy of TUCAR-T ex vivo and in vivo and we found for the first time that HLA-II deficiency could facilitate the persistence of CAR-T. Contrastively, as the most commonly eliminated target in UCAR-T, TCR deficiency was found to be a key disadvantageous factor for the shorter-term anti-tumor efficacy in vivo. Our study demonstrated ETUCAR-T could effectively resist allogeneic NK rejection ex vivo and in vivo. DISCUSSION: Our research provided a potential and effective strategy for promoting the persistence of UCAR-T cells in clinical application. And it reveals the potential key factors of the poor persistence of UCAR-T along with new insights for future development. CI - Copyright (c) 2022 Li, Zhu, Xu, Chen, Zhang, Yang, Qi, Hong, Li, Wang, Shen and Qian. FAU - Li, Wuling AU - Li W AD - Key Laboratory for Biorheological Science and Technology of Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China. AD - Center for Precision Medicine of Cancer, Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing, China. FAU - Zhu, Xiuxiu AU - Zhu X AD - Key Laboratory for Biorheological Science and Technology of Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China. AD - Center for Precision Medicine of Cancer, Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing, China. FAU - Xu, Yanmin AU - Xu Y AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Chen, Jun AU - Chen J AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Zhang, Hongtao AU - Zhang H AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Yang, Zhi AU - Yang Z AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Qi, Yanan AU - Qi Y AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Hong, Juan AU - Hong J AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Li, Yunyan AU - Li Y AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Wang, Guixue AU - Wang G AD - Key Laboratory for Biorheological Science and Technology of Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China. AD - Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, Bioengineering College of Chongqing University, Chongqing, China. FAU - Shen, Junjie AU - Shen J AD - Chongqing Key Laboratory of Gene and Cell Therapy, Institute of Precision Medicine and Biotechnology, Chongqing Precision Biotech Co., Ltd., Chongqing, China. FAU - Qian, Cheng AU - Qian C AD - Key Laboratory for Biorheological Science and Technology of Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China. AD - Center for Precision Medicine of Cancer, Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20221202 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - 0 (Receptors, Chimeric Antigen) RN - 0 (Receptors, Antigen, T-Cell) RN - 0 (Histocompatibility Antigens Class I) RN - 0 (HLA Antigens) RN - 0 (Histocompatibility Antigens Class II) SB - IM MH - Humans MH - Mice MH - Animals MH - *Receptors, Chimeric Antigen/genetics MH - Receptors, Antigen, T-Cell/genetics MH - Histocompatibility Antigens Class I MH - HLA Antigens/genetics MH - *Neoplasms MH - Histocompatibility Antigens Class II MH - HLA-E Antigens PMC - PMC9757162 OTO - NOTNLM OT - CRISPR/Cas9 OT - ETUCAR-T OT - HLA-E OT - natural killer cell OT - universal CAR COIS- Chongqing Precision Biotech Co., Ltd. is a biotechnology company focused on research and development of tumor cellular immunotherapy. JS, YX, HZ, JC, ZY, YQ, JH, YL, are full-time employees of Chongqing Precision Biotech Co., Ltd. CQ is the chief scientist of this company. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2022/12/20 06:00 MHDA- 2022/12/21 06:00 PMCR- 2022/01/01 CRDT- 2022/12/19 04:04 PHST- 2022/09/24 00:00 [received] PHST- 2022/11/14 00:00 [accepted] PHST- 2022/12/19 04:04 [entrez] PHST- 2022/12/20 06:00 [pubmed] PHST- 2022/12/21 06:00 [medline] PHST- 2022/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2022.1052717 [doi] PST - epublish SO - Front Immunol. 2022 Dec 2;13:1052717. doi: 10.3389/fimmu.2022.1052717. eCollection 2022.