mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic Leukemia

Abstract Clinical trials of Chimeric Antigen Receptor T‐cell (CAR‐T) therapy have demonstrated remarkable success in treating both solid tumors and hematological malignancies. Nanobodies (Nbs) have emerged as promising antigen‐targeting domains for CARs, owing to their high specificity, robust stabi...

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Main Authors: Zhixiong Zhu, Hexian Li, Qizhong Lu, Zongliang Zhang, Jia Li, Zeng Wang, Nian Yang, Zhengyu Yu, Chen Yang, Yongdong Chen, Huaqing Lu, Wei Wang, Ting Niu, Chunlai Nie, Aiping Tong
Format: Article
Language:English
Published: Wiley 2024-09-01
Series:Advanced Science
Subjects:
Online Access:https://doi.org/10.1002/advs.202400024
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author Zhixiong Zhu
Hexian Li
Qizhong Lu
Zongliang Zhang
Jia Li
Zeng Wang
Nian Yang
Zhengyu Yu
Chen Yang
Yongdong Chen
Huaqing Lu
Wei Wang
Ting Niu
Chunlai Nie
Aiping Tong
author_facet Zhixiong Zhu
Hexian Li
Qizhong Lu
Zongliang Zhang
Jia Li
Zeng Wang
Nian Yang
Zhengyu Yu
Chen Yang
Yongdong Chen
Huaqing Lu
Wei Wang
Ting Niu
Chunlai Nie
Aiping Tong
author_sort Zhixiong Zhu
collection DOAJ
description Abstract Clinical trials of Chimeric Antigen Receptor T‐cell (CAR‐T) therapy have demonstrated remarkable success in treating both solid tumors and hematological malignancies. Nanobodies (Nbs) have emerged as promising antigen‐targeting domains for CARs, owing to their high specificity, robust stability, and strong affinity, leading to significant advancements in the field of Nb‐CAR‐T. In the realm of T‐cell acute lymphoblastic leukemia (T‐ALL) targets, CD5 stands out as a potentially excellent candidate for T‐cell‐based CAR therapy, due to its distinct expression on the surface of malignant T‐ALL cells. To mitigate graft‐versus‐host disease associated with allogeneic CAR‐T, γδT cells are selected and stimulated from peripheral blood mononuclear cells, and γδT cells are engineered via CRISPR/Cas9 to eliminate fratricide, enabling the creation of fratricide‐resistant CAR‐γδTCD5− cells. In vitro transcribed (IVT) mRNA is used to construct CAR‐T, presenting a safer, faster, and cost‐effective method compared to traditional viral vector approaches. In this study, a CD5‐VHH library is constructed, and specific CD5‐nanobodies are screened for subsequent use in CD5‐CAR‐γδTCD5− therapy. IVT‐mRNA‐CD5‐CAR‐γδTCD5− cells exhibited favorable functional characteristics and demonstrated antitumor efficacy against malignant T cell lines, underlining the potential for advancing mRNA‐CD5‐CAR‐γδTCD5− therapy.
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spelling doaj-art-8b15ce605dcc484b9a66e75fbf9aa14f2025-08-20T01:55:16ZengWileyAdvanced Science2198-38442024-09-011135n/an/a10.1002/advs.202400024mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic LeukemiaZhixiong Zhu0Hexian Li1Qizhong Lu2Zongliang Zhang3Jia Li4Zeng Wang5Nian Yang6Zhengyu Yu7Chen Yang8Yongdong Chen9Huaqing Lu10Wei Wang11Ting Niu12Chunlai Nie13Aiping Tong14State Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaDepartment of Hematology State Key Laboratory of Biotherapy and Cancer Center West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaDepartment of Hematology State Key Laboratory of Biotherapy and Cancer Center West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaState Key Laboratory of Biotherapy and Cancer Center Collaborative Innovation Center of Biotherapy West China Hospital Sichuan University Chengdu 610041 ChinaAbstract Clinical trials of Chimeric Antigen Receptor T‐cell (CAR‐T) therapy have demonstrated remarkable success in treating both solid tumors and hematological malignancies. Nanobodies (Nbs) have emerged as promising antigen‐targeting domains for CARs, owing to their high specificity, robust stability, and strong affinity, leading to significant advancements in the field of Nb‐CAR‐T. In the realm of T‐cell acute lymphoblastic leukemia (T‐ALL) targets, CD5 stands out as a potentially excellent candidate for T‐cell‐based CAR therapy, due to its distinct expression on the surface of malignant T‐ALL cells. To mitigate graft‐versus‐host disease associated with allogeneic CAR‐T, γδT cells are selected and stimulated from peripheral blood mononuclear cells, and γδT cells are engineered via CRISPR/Cas9 to eliminate fratricide, enabling the creation of fratricide‐resistant CAR‐γδTCD5− cells. In vitro transcribed (IVT) mRNA is used to construct CAR‐T, presenting a safer, faster, and cost‐effective method compared to traditional viral vector approaches. In this study, a CD5‐VHH library is constructed, and specific CD5‐nanobodies are screened for subsequent use in CD5‐CAR‐γδTCD5− therapy. IVT‐mRNA‐CD5‐CAR‐γδTCD5− cells exhibited favorable functional characteristics and demonstrated antitumor efficacy against malignant T cell lines, underlining the potential for advancing mRNA‐CD5‐CAR‐γδTCD5− therapy.https://doi.org/10.1002/advs.202400024CAR‐TIVT‐mRNAT‐ALLγδT cells
spellingShingle Zhixiong Zhu
Hexian Li
Qizhong Lu
Zongliang Zhang
Jia Li
Zeng Wang
Nian Yang
Zhengyu Yu
Chen Yang
Yongdong Chen
Huaqing Lu
Wei Wang
Ting Niu
Chunlai Nie
Aiping Tong
mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic Leukemia
Advanced Science
CAR‐T
IVT‐mRNA
T‐ALL
γδT cells
title mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic Leukemia
title_full mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic Leukemia
title_fullStr mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic Leukemia
title_full_unstemmed mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic Leukemia
title_short mRNA‐Engineered CD5‐CAR‐γδTCD5‐ Cells for the Immunotherapy of T‐Cell Acute Lymphoblastic Leukemia
title_sort mrna engineered cd5 car γδtcd5 cells for the immunotherapy of t cell acute lymphoblastic leukemia
topic CAR‐T
IVT‐mRNA
T‐ALL
γδT cells
url https://doi.org/10.1002/advs.202400024
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