Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancer

Abstract Purpose Circular RNA (circRNA) has emerged as a promising RNA therapeutic molecule due to its enhanced stability and prolonged protein expression compared to messenger RNA (mRNA). Using circRNA to construct transient Chimeric Antigen Receptor (CAR)-T cells can mitigate the limitations of co...

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Main Authors: Jingsheng Cai, Zheng Liu, Shaoyi Chen, Jingwei Zhang, Haoran Li, Xun Wang, Feng Yang, Shaodong Wang, Xiao Li, Yun Li, Kezhong Chen, Jun Wang, Ming Sun, Mantang Qiu
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Experimental Hematology & Oncology
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Online Access:https://doi.org/10.1186/s40164-025-00625-8
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author Jingsheng Cai
Zheng Liu
Shaoyi Chen
Jingwei Zhang
Haoran Li
Xun Wang
Feng Yang
Shaodong Wang
Xiao Li
Yun Li
Kezhong Chen
Jun Wang
Ming Sun
Mantang Qiu
author_facet Jingsheng Cai
Zheng Liu
Shaoyi Chen
Jingwei Zhang
Haoran Li
Xun Wang
Feng Yang
Shaodong Wang
Xiao Li
Yun Li
Kezhong Chen
Jun Wang
Ming Sun
Mantang Qiu
author_sort Jingsheng Cai
collection DOAJ
description Abstract Purpose Circular RNA (circRNA) has emerged as a promising RNA therapeutic molecule due to its enhanced stability and prolonged protein expression compared to messenger RNA (mRNA). Using circRNA to construct transient Chimeric Antigen Receptor (CAR)-T cells can mitigate the limitations of conventional viral vector-based CAR-T approaches, such as complex process and long-term side effects. Methods The study first reconfirmed the advantageous properties of circRNA, focusing on its stability and protein expression efficiency. Electroporation conditions were then optimized for the efficient delivery of circRNA into human primary T cells. Subsequently, a circRNA encoding the anti-Delta-like Ligand 3 (DLL3) CAR was constructed, and CAR-T cells were generated via electroporation. The efficacy of circRNA-based CAR-T cells was compared to mRNA-based CAR-T cells in both in vitro and in vivo models, including subcutaneous and orthotopic small cell lung cancer (SCLC) mouse models. Results CircRNA-based CAR-T cells demonstrated superior efficacy against SCLC compared to mRNA-based CAR-T cells. In vitro experiments showed enhanced tumor-killing effects, while in vivo studies revealed complete elimination of human SCLC tumors in both subcutaneous and orthotopic mouse models. These results underscored the therapeutic advantages of circRNA in CAR-T cell therapy. Conclusions This study validated the feasibility of the circRNA-electroporation strategy in CAR-T cell therapy and offered a potentially effective approach for treating SCLC, highlighting the potential of circRNA-based technologies in advancing cell therapies. Graphic Abstract
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spelling doaj-art-e866d84d94574b02b4c482829685c17b2025-08-20T03:02:18ZengBMCExperimental Hematology & Oncology2162-36192025-03-0114111810.1186/s40164-025-00625-8Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancerJingsheng Cai0Zheng Liu1Shaoyi Chen2Jingwei Zhang3Haoran Li4Xun Wang5Feng Yang6Shaodong Wang7Xiao Li8Yun Li9Kezhong Chen10Jun Wang11Ming Sun12Mantang Qiu13Thoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalDepartment of Thoracic Surgery, Sichuan Clinical Research Center for Cancer, Sichuan Cancer Hospital& Institute, Sichuan Cancer Center, University of Electronic Science and Technology of China (UESTC)Thoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalThoracic Oncology Institute, Peking University People’s HospitalDepartment of Oncology Center, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal HospitalThoracic Oncology Institute, Peking University People’s HospitalAbstract Purpose Circular RNA (circRNA) has emerged as a promising RNA therapeutic molecule due to its enhanced stability and prolonged protein expression compared to messenger RNA (mRNA). Using circRNA to construct transient Chimeric Antigen Receptor (CAR)-T cells can mitigate the limitations of conventional viral vector-based CAR-T approaches, such as complex process and long-term side effects. Methods The study first reconfirmed the advantageous properties of circRNA, focusing on its stability and protein expression efficiency. Electroporation conditions were then optimized for the efficient delivery of circRNA into human primary T cells. Subsequently, a circRNA encoding the anti-Delta-like Ligand 3 (DLL3) CAR was constructed, and CAR-T cells were generated via electroporation. The efficacy of circRNA-based CAR-T cells was compared to mRNA-based CAR-T cells in both in vitro and in vivo models, including subcutaneous and orthotopic small cell lung cancer (SCLC) mouse models. Results CircRNA-based CAR-T cells demonstrated superior efficacy against SCLC compared to mRNA-based CAR-T cells. In vitro experiments showed enhanced tumor-killing effects, while in vivo studies revealed complete elimination of human SCLC tumors in both subcutaneous and orthotopic mouse models. These results underscored the therapeutic advantages of circRNA in CAR-T cell therapy. Conclusions This study validated the feasibility of the circRNA-electroporation strategy in CAR-T cell therapy and offered a potentially effective approach for treating SCLC, highlighting the potential of circRNA-based technologies in advancing cell therapies. Graphic Abstracthttps://doi.org/10.1186/s40164-025-00625-8Circular RNAElectroporationCAR-TDelta-like Ligand 3Small cell lung cancer
spellingShingle Jingsheng Cai
Zheng Liu
Shaoyi Chen
Jingwei Zhang
Haoran Li
Xun Wang
Feng Yang
Shaodong Wang
Xiao Li
Yun Li
Kezhong Chen
Jun Wang
Ming Sun
Mantang Qiu
Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancer
Experimental Hematology & Oncology
Circular RNA
Electroporation
CAR-T
Delta-like Ligand 3
Small cell lung cancer
title Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancer
title_full Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancer
title_fullStr Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancer
title_full_unstemmed Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancer
title_short Engineered circular RNA-based DLL3-targeted CAR-T therapy for small cell lung cancer
title_sort engineered circular rna based dll3 targeted car t therapy for small cell lung cancer
topic Circular RNA
Electroporation
CAR-T
Delta-like Ligand 3
Small cell lung cancer
url https://doi.org/10.1186/s40164-025-00625-8
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